Osteoclasts are essential for maintaining healthy bone.Pathological elevation of os-teoclastogenesis or osteoclast activity can cause osteoporosis and increase the risk of bone fracture.However,a few options are avail...Osteoclasts are essential for maintaining healthy bone.Pathological elevation of os-teoclastogenesis or osteoclast activity can cause osteoporosis and increase the risk of bone fracture.However,a few options are available for directly measuring osteoclast activity in vivo to test interventions that may affect osteoclasts.Here,we describe an in vivo method to measure osteoclast-mediated bone loss targeted at normal mouse calvaria.The method employs a novel procedure for measuring osteoclast resorption pits using micro-computed tomography.The potential utility of this mouse calvaria model to assess therapies targeting osteoclasts was validated using zoledronic acid,which is a nitrogen-containing bisphosphonate drug used to treat osteoporosis.展开更多
Alzheimer’s disease is the most common cause of dementia.Although increasing evidence suggests that disruptions in lipid metabolism are closely associated with the disease,the overall profile of lipid and sterol chan...Alzheimer’s disease is the most common cause of dementia.Although increasing evidence suggests that disruptions in lipid metabolism are closely associated with the disease,the overall profile of lipid and sterol changes that occur in the brain during Alzheimer’s disease remains unclear.In this study,we compared brain tissues extracted from 32-week-old male wild-type mice and 5×FAD transgenic Alzheimer’s disease model mice,which carry mutations in the amyloid precursor protein(APP)and presenilin 1(PS1)genes.Using untargeted lipidomics and sterolomics techniques,we investigated the metabolic profiles of lipids,with a focus on sterols specifically,in three brain regions:cerebellum,hippocampus,and olfactory bulb.Our results revealed significant alterations in various lipids,particularly in the hippocampus and olfactory bulb,suggesting changes in energy levels in these regions.Further pathway analysis indicated notable disruptions in key metabolic processes,particularly those related to fatty acids and cell membrane components.Additionally,we observed decreased expression of 15 genes involved in lipid and sterol regulation.Collectively,these findings provide new insights into how imbalances in lipid and sterol metabolism may contribute to the progression of Alzheimer’s disease,highlighting potential metabolic pathways involved in the development of this debilitating disease.展开更多
Realistic models for cancer research representing disease progression that commensurately respond to therapeutics consistent with clinical observation are the holy grail for pre-clinical research and screening.Althoug...Realistic models for cancer research representing disease progression that commensurately respond to therapeutics consistent with clinical observation are the holy grail for pre-clinical research and screening.Although such an ideal is elusive,well-characterized in vivo models facilitate our understanding of disease,progression,and therapeutic opportunities.Here,we characterize a commonly used syngeneic BALB/c mouse model of triple negative breast cancer(4T1)after establishing tumors in their flanks.Tumors developed at the subcutaneous injection site for all experimental mice and their volumes were monitored.We quantified a rare subset of breast cancer stemlike cells(CSCs),classified as CD44^(+)/CD24^(−)phenotypes in in vitro and ex vivo cell populations.Chromosome numbers in ex vivo metaphase cells were greater than cells cultured in vitro(89.4±3.4,range of 70-132 and 82.6±1.1,range of 70-128;respectively).Further,we observed different types of chromosome aberrations,including gap,deletion,exchange,interstitial deletion,terminal deletion,ring,dicentric,and Robertsonian translocations.For both sources of cells,the number of aberrations was dominated by deletions,terminal deletions,and Robertsonian translocations.Ex vivo cells exhibited greater prevalence of deletions and terminal deletions,whereas in vitro cells displayed more ring aberrations and Robertsonian translocations.In conclusion,we successfully characterized cancer cells from a syngeneic mouse model of breast cancer in terms of rare CSC proportion and a variety of chromosomal aberrations,which is useful for understanding tumor traits associated with cancer development and therapeutic action.The data act as a valuable resource for other studies using the 4T1 BALB/c model.展开更多
The brain's functions are governed by molecular metabolic networks.However,due to the sophisticated spatial organization and diverse activities of the brain,characterizing both the minute and large-scale metabolic...The brain's functions are governed by molecular metabolic networks.However,due to the sophisticated spatial organization and diverse activities of the brain,characterizing both the minute and large-scale metabolic activity across the entire brain and its numerous micro-regions remains incredibly challenging.Here,we offer a high-definition spatially resolved metabolomics technique to better understand the metabolic specialization and interconnection throughout the mouse brain using improved ambient mass spectrometry imaging.This method allows for the simultaneous mapping of thousands of metabolites at a 30 μm spatial resolution across the mouse brain,ranging from structural lipids to functional neurotransmitters.This approach effectively reveals the distribution patterns of delicate microregions and their distinctive metabolic characteristics.Using an integrated database,we annotated 259 metabolites,demonstrating that the metabolome and metabolic pathways are unique to each brain microregion.The distribution of metabolites,closely linked to functionally connected brain regions and their interactions,offers profound insights into the complexity of chemical processes and their roles in brain function.An initial dataset for future metabolomics research might be obtained from the high-definition mouse brain's spatial metabolome atlas.展开更多
Background:In recent decades,the global incidence of dengue fever has been stead-ily increasing,with continuous geographical expansion.Researchers have successfully modeled most clinical symptoms of human dengue fever...Background:In recent decades,the global incidence of dengue fever has been stead-ily increasing,with continuous geographical expansion.Researchers have successfully modeled most clinical symptoms of human dengue fever using interferon type I(IFN-I)or combined IFN-I/II receptor knockout mice infected with dengue virus(DENV).However,this model requires further optimization to better support related studies.Methods:This study aimed to establish a stable dengue infection model by evaluating the effects of different genetic backgrounds and injection routes on DENV infection in interferon receptor knockout mice.We first infected various strains of interferon receptor-deficient mice with DENV and compared their susceptibility based on clini-cal symptoms,viremia levels,organ indices,histopathological findings,and vascular leakage markers.Subsequently,we selected the most susceptible strain to further investigate the impact of different injection methods on infection outcomes.Results:We found that BALB/c background mice with type 1 interferon recep-tor knockout(IFNAR)had the most obvious symptoms.Subsequently,we selected IFNAR−/−BALB/c mice to further explore the effects of different injection methods on dengue virus infection.The results showed that the intraperitoneal injection group had the most severe clinical symptoms,the longest duration of viremia,and the most obvious degree of organ damage.Conclusion:Through systematic screening and optimization,we established a robust animal model of dengue virus infection via intraperitoneal injection in IFNAR−/−BALB/c mice.This model offers a valuable tool for future dengue research.展开更多
Background:Humanized mouse models are essential for studying the human immune response and antibody development.However,conventional models show limited B cell maturation and antigen-specific humoral responses.To over...Background:Humanized mouse models are essential for studying the human immune response and antibody development.However,conventional models show limited B cell maturation and antigen-specific humoral responses.To overcome these limitations,we used the NOG-EXL mice expressing human interleukin 3(IL-3)and granulocyte-macrophage colony-stimulating factor(GM-CSF)to enhance myeloid and B-cell lineage differentiation.Methods:Human CD34^(+)hematopoietic stem cells(HSC)were transplanted into NOG-EXL mice to produce humanized immune systems.After immune cell reconstitution was confirmed across 12 weeks,the mice were immunized twice with inactivated severe fever with thrombocytopenia syndrome virus(SFTSV)antigens.Peripheral blood mononuclear cells and splenocytes were analyzed using multicolor flow cytometry to assess human immune cell subsets.Antigen-specific immunoglobulin G(IgG)production was quantified using enzyme-linked immunosorbent assay(ELISA),and virus-specific B cells were isolated using antigen-labeled recombinant protein probes.Results:Twelve weeks after transplantation of HSCs into NOG-EXL mice,they exhibited robust engraftment of human leukocytes,including T,B,and dendritic cells,compared to NOG mice.Unlike NOG mice,humanized NOG-EXL mice exhibited an increase in human IgG levels,indicating the production of human antibody responses to antigens.Humanized NOG-EXL mice were immunized twice every 2 weeks with inactivated SFTSV,and antigen-specific human antibodies against the virus were detected in the mouse sera by ELISA.Sera from SFTSV-immunized humanized mice demonstrated neutralizing activity against SFTSV,confirming the induction of functional virus-specific neutralizing antibodies.Antigen-binding IgG-positive human B cells were isolated from mouse splenocytes using recombinant protein probes.Conclusion:This model provides a valuable platform for evaluating humoral immunity and isolating B cells using high-affinity human monoclonal antibodies without genetic engineering.展开更多
Background:KIT proto-oncogene,receptor tyrosine kinase(KIT,CD117)and platelet-derived growth factor-alpha(PDGFRA)are key drivers of gastrointestinal stromal tumors(GIST),but resistance to targeted therapy often arises...Background:KIT proto-oncogene,receptor tyrosine kinase(KIT,CD117)and platelet-derived growth factor-alpha(PDGFRA)are key drivers of gastrointestinal stromal tumors(GIST),but resistance to targeted therapy often arises from tumor protein p53(p53)alterations and loss of cell cycle control.However,the role of p53 status in GIST therapeutic potential has rarely been studied,so this study aimed to employ both wild-type andmutant p53 GIST models to investigate how p53 dysfunction influences the efficacy of p53 pathway-targeted therapies.Methods:The efficacy of the mouse double minute 2 homolog(MDM2)inhibitor(HDM201)and the Wee1 G2 checkpoint kinase(Wee1)inhibitor(adavosertib)was confirmed in both p53 wild-type(p53 WT)and p53 mutant(p53 MT)GIST cells.The anti-proliferative effects were assessed using the Cell Counting Kit-8(CCK-8)assay.Flow cytometry(FACS)and immunoblotting were employed to evaluate apoptosis and the expression of proteins related to drug efficacy.These findings were further validated in a xenograft model.Results:HDM201 selectively inhibited growth and triggered apoptosis in p53WT GIST cells,while adavosertib was effective mainly in p53 MT cells.Western blot analysis revealed thatHDM201 increased p53 and p21 levels in p53WT cells,and adavosertib affectedWee1 and phospho-cdc2 expression in both p53WT and p53 MT cells.In a xenograft mouse model,HDM201 significantly reduced the tumor volume and weight in p53WTGIST cells,whereas p53MT tumors showed only a moderate size reduction with adavosertib,without significant changes.Conclusions:Our results highlight the importance of p53 status in guiding GIST treatment.p53 WT tumors respond toMDM2 inhibitors,while p53 MTtumors show greater sensitivity toWee1 inhibitors,supporting p53 pathway targeting as a promising strategy for GIST patients.展开更多
Epithelial ovarian cancer(EOC) is the leading cause of gynecological cancer-related mortality in the developed world. EOC is a heterogeneous disease represented by several histological and molecular subtypes. Therefor...Epithelial ovarian cancer(EOC) is the leading cause of gynecological cancer-related mortality in the developed world. EOC is a heterogeneous disease represented by several histological and molecular subtypes. Therefore, exploration of relevant preclinical animal models that consider the heterogenic nature of EOC is of great importance for the development of novel therapeutic strategies that can be translated clinically to combat this devastating disease. In this review, we discuss recent progress in the development of preclinical mouse models for EOC study as well as their advantages and limitations.展开更多
Genetically engineered mouse(GEM)models are commonly used in biomedical research.Generating GEMs involve complex set of experimental procedures requiring sophisticated equipment and highly skilled technical staff.Beca...Genetically engineered mouse(GEM)models are commonly used in biomedical research.Generating GEMs involve complex set of experimental procedures requiring sophisticated equipment and highly skilled technical staff.Because of these reasons,most research institutes set up centralized core facilities where custom GEMs are created for research groups.Researchers,on the other hand,when they begin thinking about generating GEMs for their research,several questions arise in their minds.For example,what type of model(s)would be best useful for my research,how do I design them,what are the latest technologies and tools available for developing my model(s),and finally how to breed GEMs in my research.As there are several considerations and options in mouse designs,and as it is an expensive and time-consuming endeavor,careful planning upfront can ensure the highest chance of success.In this article,we provide brief answers to several frequently asked questions that arise when researchers begin thinking about generating mouse model(s)for their work.展开更多
This paper introduces part of the content in the association standard,T/CAAM0002–2020 Nomenclature and Location of Acupuncture Points for Laboratory Animals Part 3:Mouse.This standard was released by the China Associ...This paper introduces part of the content in the association standard,T/CAAM0002–2020 Nomenclature and Location of Acupuncture Points for Laboratory Animals Part 3:Mouse.This standard was released by the China Association of Acupuncture and Moxibustion on May 15,2020,implemented on October 31,2020,and published by Standards Press of China.The standard was drafted by the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,and the Nanjing University of Chinese Medicine.Principal draftsmen:Xiang-hong JING and Xing-bang HUA.Participating draftsmen:Wan-zhu BAI,Bin XU,Dong-sheng XU,Yi GUO,Tie-ming MA,Xin-jun WANG,and Sheng-feng LU.展开更多
Background:Over the past few decades,a threefold increase in obesity and type 2 diabetes(T2D)has placed a heavy burden on the health-care system and society.Previous studies have shown correlations between obesity,T2D...Background:Over the past few decades,a threefold increase in obesity and type 2 diabetes(T2D)has placed a heavy burden on the health-care system and society.Previous studies have shown correlations between obesity,T2D,and neurodegenera-tive diseases,including dementia.It is imperative to further understand the relation-ship between obesity,T2D,and cognitive deficits.Methods:This investigation tested and evaluated the cognitive impact of obesity and T2D induced by high-fat diet(HFD)and the effect of the host genetic background on the severity of cognitive decline caused by obesity and T2D in collaborative cross(CC)mice.The CC mice are a genetically diverse panel derived from eight inbred strains.Results:Our findings demonstrated significant variations in the recorded phenotypes across different CC lines compared to the reference mouse line,C57BL/6J.CC037 line exhibited a substantial increase in body weight on HFD,whereas line CC005 ex-hibited differing responses based on sex.Glucose tolerance tests revealed significant variations,with some lines like CC005 showing a marked increase in area under the curve(AUC)values on HFD.Organ weights,including brain,spleen,liver,and kidney,varied significantly among the lines and sexes in response to HFD.Behavioral tests using the Morris water maze indicated that cognitive performance was differentially affected by diet and genetic background.Conclusions:Our study establishes a foundation for future quantitative trait loci map-ping using CC lines and identifying genes underlying the comorbidity of Alzheimer's disease(AD),caused by obesity and T2D.The genetic components may offer new tools for early prediction and prevention.展开更多
Background:Most mutations in the COL6A3 gene lead to collagen VI-related myopathies.This is due to a reduced expression or mislocalization of the COL6A3 protein.Therefore,studying the consequence of knocking out the C...Background:Most mutations in the COL6A3 gene lead to collagen VI-related myopathies.This is due to a reduced expression or mislocalization of the COL6A3 protein.Therefore,studying the consequence of knocking out the Col6a3 gene in mouse models is relevant,but the Col6a3 mouse models reported so far do not entirely abolish COL6A3 protein expression.Methods:Here,we present the development,validation and preliminary phenotypic characterization of a novel CRISPR-based knockout mouse model targeting Col6a3 exon 3(Col6a3^(d3/d3)).Results:In this mouse model,Col6a3 mRNA is still expressed at a similar level to wild-type littermates,although the expected protein is undetectable by mass spectrometry.Histological analysis of Col6a3^(d3/d3)quadriceps revealed an abnormally high frequency of muscle cells with internally nucleated muscle cells,consistent with a myopathy phenotype.Interestingly,Col6a3^(d3/d3)mice are smaller in size,with their fat,muscle,and bone kept proportional compared to wild-type littermates.Conclusions:In summary,we performed the validation and preliminary phenotypic characterization of a novel Col6a3 knockout mouse model that could be further characterized and used to study COL6A3 biology and model collagen VI-associated diseases.展开更多
Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific...Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific animal models induced by inflammatory cytokines.This study introduces a novel mouse model of PD driven by the proinflammatory cytokine CXCL1,identified in our previous research.The involvement of CXCL1 in PD pathogenesis was validated using subacute and chronic MPTP-induced mouse models.Based on these findings,2-month-old C57BL/6J mice were intravenously administered CXCL1(20 ng/kg/day)for 2 weeks(5 days per week),successfully replicating motor deficits and pathological alterations in the substantia nigra observed in the chronic MPTP model.These results demonstrate the potential of CXCL1-induced inflammation as a mechanism for PD modeling.The model revealed activation of the PPAR signaling pathway in CXCL1-mediated neuronal damage by CXCL1.Linoleic acid,a PPAR-γactivator,significantly mitigated MPTPand CXCL1-induced toxicity and reduced serum CXCL1levels.In addition,the CXCL1-injected mouse model shortened the timeline for developing chronic PD mouse model to 2 weeks,offering an efficient platform for studying inflammation-driven processes in PD.The findings provide critical insights into the inflammatory mechanisms underlying PD and identify promising therapeutic targets for intervention.展开更多
Alzheimer'sdisease(AD)isaprogressive neurodegenerative disorder characterized by cognitive impairment and distinct neuropathological features,including amyloid-βplaques,neurofibrillary tangles,and reactive astrog...Alzheimer'sdisease(AD)isaprogressive neurodegenerative disorder characterized by cognitive impairment and distinct neuropathological features,including amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis.Developing effective diagnostic,preventative,and therapeutic strategies for AD necessitates the establishment of animal models that accurately recapitulate the pathophysiological processes of the disease.Existing transgenic mouse models have significantly contributed to understanding AD pathology but often fail to replicate the complexity of human AD.Additionally,these models are limited in their ability to elucidate the interplay among amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis due to the absence of spatially and temporally specific genetic manipulation.In this study,we introduce a novel AD mouse model(APP/PS1-TauP301L-Adeno mice)designed to rapidly induce pathological symptoms and enhance understanding of AD mechanisms.Neurofibrillary tangles and severe reactive astrogliosis were induced by injecting AAVDJ-EF1a-hTauP301L-EGFP and Adeno-GFAP-GFP viruses into the hippocampi of 5-month-old APP/PS1 mice.Three months post-injection,these mice exhibited pronounced astrogliosis,substantial amyloid-βplaque accumulation,extensiveneurofibrillarytangles,accelerated neuronal loss,elevated astrocytic GABA levels,and significant spatial memory deficits.Notably,these pathological features were less severe in AAVTauP301L-expressing APP/PS1 mice without augmented reactive astrogliosis.These findings indicate an exacerbating role of severe reactive astrogliosis in amyloid-βplaque and neurofibrillary tangle-associated pathology.The APP/PS1-TauP301L-Adeno mouse model provides a valuable tool for advancing therapeutic research aimed at mitigating the progression of AD.展开更多
Age-related osteoporosis poses a significant challenge in musculoskeletal health;a condition characterized by reduced bone density and increased fracture susceptibility in older individuals necessitates a better under...Age-related osteoporosis poses a significant challenge in musculoskeletal health;a condition characterized by reduced bone density and increased fracture susceptibility in older individuals necessitates a better understanding of underlying molecular and cellular mechanisms.Emerging evidence suggests that osteocytes are the pivotal orchestrators of bone remodeling and represent novel therapeutic targets for age-related bone loss.Our study uses the prematurely aged PolgD257A/D257A(PolgA)mouse model to scrutinize age-and sex-related alterations in musculoskeletal health parameters(frailty,grip strength,gait data),bone and particularly the osteocyte lacuno-canalicular network(LCN).Moreover,a new quantitative in silico image analysis pipeline is used to evaluate the alterations in the osteocyte network with aging.Our findings underscore the pronounced degenerative changes in the musculoskeletal health parameters,bone,and osteocyte LCN in PolgA mice as early as 40 weeks,with more prominent alterations evident in aged males.Our findings suggest that the PolgA mouse model serves as a valuable model for studying the cellular mechanisms underlying age-related bone loss,given the comparable aging signs and age-related degeneration of the bone and the osteocyte network observed in naturally aging mice and elderly humans.展开更多
Background:Glucocorticoids are used as anti-inflammatory drugs for the treatment of various diseases,however,their side effects on normal brain tissue remain underinvestigated.Objectives:The study aimed to investigate...Background:Glucocorticoids are used as anti-inflammatory drugs for the treatment of various diseases,however,their side effects on normal brain tissue remain underinvestigated.Objectives:The study aimed to investigate dexamethasone(DXM)effects on cell composition and myelin content in the mouse brain tissue.Methods:C57Bl/6 male mice(n 60)received single and ten multiple intraperitoneal DXM injections(2.5 mg/kg),and the studied=parameters were analysed at 1,3,7,10 days after a single DXM injection and 15,30,60,and 90 days after the multiple injections.Oligodendrocytes,microglia,and astrocytes were assayed by immunohistochemistry with specific antibodies(Olig2,CD68,and GFAP,respectively)in the corpus callosum of the normal brain tissue.The myelin content was estimated by staining with LuxolFastBlue.The presence of GFAP isoforms was determined by western blotting.Results:DXM administration did not affect oligodendrocytes in the mouse brain but temporarily significantly decreased myelin content(1.2-fold,p 0.0058;1.4-fold,p 0.0001)at 3–15 days time points.At the same time,DXM significantly=<decreased the number of microglial cells(1.5–3.5-fold,p 0.0001)and significantly increased astrocytes(1.8-fold,p<<0.0001).Prolonged administration of DXM resulted in the decrease of the main GFAPα-isoform(50 kDa)and the appearance of shorter GFAP isoforms(30 kDa,42 kDa,44 kDa)similar to that in some neurodegenerative animal models.Conclusion:DXM can modify the cell composition of the normal mouse brain tissue by decreasing microglial cells and increasing astrocytes.Long-term use of DXM results in the inhibition of myelin formation and the appearance of truncated GFAP isoforms,suggesting its ability to induce neurodegeneration-like changes in the normal mouse brain.展开更多
Background:Spinocerebellar ataxia type 2(SCA2)is a neurodegenerative disease marked by significant clinical and genetic heterogeneity,primarily caused by expanded CAG mutations in the ATXN2 gene.The unstable expansion...Background:Spinocerebellar ataxia type 2(SCA2)is a neurodegenerative disease marked by significant clinical and genetic heterogeneity,primarily caused by expanded CAG mutations in the ATXN2 gene.The unstable expansion of CAG repeats disrupts the genetic stability of animal models,which is detrimental to disease research.Methods:In this study,we established a mouse model in which CAG repeats do not undergo microsatellite instability(MSI)across generations.A humanized ATXN2 cDNA with four CAA interruptions within 73 CAG expansions was inserted into the Rosa26 locus of C57BL/6J mice.A 23 CAG control mouse model was also generated to verify ATXN2 integration and expression.Results:In our model,the number of CAG repeats remained stable during transmission,with no CAG repeat expansion observed in 64 parent-to-offspring transmissions.Compared with SCA2-Q23 mice,SCA2-Q73 mice exhibited progressive motor impairment,reduced Purkinje cell count and volume(indicative of cell atrophy),and muscle atrophy.These observations in the mice suggest that the behavioral and neuropathological phenotypes may reflect the features of SCA2 patients.RNA-seq analysis of the gastrocnemius muscle in SCA2-Q73 mice showed significant changes in muscle differentiation and development gene expression at 56 weeks,with no significant differences at 16 weeks compared to SCA2-Q23 mice.The expression level of the Myf6 gene significantly changed in the muscles of aged mice.Conclusion:In summary,the establishment of this model not only provides a stable animal model for studying CAG transmission in SCA2 but also indicates that the lack of long-term neural stimulation leads to muscle atrophy.展开更多
The Leprdb/db mouse is a common and well-studied model of type II diabetes mel-litus that is often employed in biomedical research.Despite being one of the most commonly used models for the investigation of diabetic w...The Leprdb/db mouse is a common and well-studied model of type II diabetes mel-litus that is often employed in biomedical research.Despite being one of the most commonly used models for the investigation of diabetic wound healing,there are a few specific guidelines for its husbandry,and wound complications such as infection and expansion are common.This study presents a modified animal husbandry ap-proach for the Leprdb/db mouse to reduce the incidence of complications during wound healing experiments.Compared to standard rodent housing protocols,the use of this modified protocol leads to decreased rates of complications among experimental animals across several experiments.The protocol includes increased cage size,de-creased housing density,and more frequent cage replacements.The use of improved husbandry for the Leprdb/db mouse decreases the total number of animals required,minimizes harm during experimentation,and improves the consistency and reproduc-ibility of wound healing studies.展开更多
Dear Editor,Crimean–Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a severe tick-borne illness with a wide geographical distribution,posing a significant threat with case fatality rates ranging from...Dear Editor,Crimean–Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a severe tick-borne illness with a wide geographical distribution,posing a significant threat with case fatality rates ranging from 5%to 70%(Hawman and Feldmann,2023).Due to the lack of approved vaccines and therapeutics,the World Health Organization(WHO)has listed CCHF as one of the priority diseases(Semper et al.,2024).CCHF initially presents as a nonspecific febrile illness,characterized by fever,malaise,myalgia,and nausea,which can rapidly progress to hemorrhagic disease.The hemorrhagic stage is particularly pronounced in severe cases,with rapid progression to disseminated intravascular coagulation(DIC),overt bleeding,kidney or liver failure,and shock(Frank et al.,2024).Up to date,there is an absence of a suitable animal model that can accurately mimic the coagulopathy and bleeding associated with CCHFV infection.Consequently,our understanding of the pathogenic mechanisms underlying these conditions remains limited(Rodriguez et al.,2022).展开更多
Background:The current understanding of diabetic kidney disease(DKD)has significant gaps regarding the underlying pathogenesis.In this study,we aimed to characterize the temporal progression of DKD using a state-of-th...Background:The current understanding of diabetic kidney disease(DKD)has significant gaps regarding the underlying pathogenesis.In this study,we aimed to characterize the temporal progression of DKD using a state-of-the-art mouse model of hypertension-accelerated disease,integrating kidney biomarker analysis,histopathology,and glomerular transcriptomic profiling.Methods:Female diabetic db/db mice received a single intravenous dose of adenoassociated virus-mediated renin overexpression(ReninAAV,week 5)and underwent uninephrectomy(UNx,week 4).db/db UNx-ReninAAV mice were terminated at weeks 1,4,8,and 12(n=7–8 per group).Female db/m mice were used as healthy controls.Study endpoints included plasma and urine biochemistry,glomerulosclerosis scoring,quantitative kidney histology,and RNA sequencing of glomeruli isolated using lasercapture microdissection.Results:db/db UNx-ReninAAV mice developed progressive albuminuria(from week 4)and glomerulosclerosis(from week 8).A pathway analysis of clustered gene regulations revealed broad glomerular transcriptome perturbations with signatures of increased extracellular matrix(ECM)turnover from week 8 and early onset of metabolic dysfunction.Markers of glomerular cell types and injury exhibited temporal regulation over the course of DKD,with early and sustained downregulation of endothelial markers,heterogeneous regulation of podocyte markers,and significant mesangial and parietal epithelial aberrations.Furthermore,the upregulation of cell injury markers confirmed progressive glomerular injury in the model.Conclusion:The db/db UNx-ReninAAV mouse model exhibits distinct temporal dynamics in glomerular cell markers,metabolic dysregulation,ECM remodeling,and injury.Together,these results highlight the utility of the db/db UNx-ReninAAV model as a relevant preclinical platform for studying progressive DKD.展开更多
基金National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health,Grant/Award Number:R01AR069044Rutgers-New Jersey Medical School Department of Orthopaedics。
文摘Osteoclasts are essential for maintaining healthy bone.Pathological elevation of os-teoclastogenesis or osteoclast activity can cause osteoporosis and increase the risk of bone fracture.However,a few options are available for directly measuring osteoclast activity in vivo to test interventions that may affect osteoclasts.Here,we describe an in vivo method to measure osteoclast-mediated bone loss targeted at normal mouse calvaria.The method employs a novel procedure for measuring osteoclast resorption pits using micro-computed tomography.The potential utility of this mouse calvaria model to assess therapies targeting osteoclasts was validated using zoledronic acid,which is a nitrogen-containing bisphosphonate drug used to treat osteoporosis.
基金supported by the National Natural Science Foundation of China,Nos.82200784,32271311Qizhen Foundation,No.226‐2023‐00008(all to LH).
文摘Alzheimer’s disease is the most common cause of dementia.Although increasing evidence suggests that disruptions in lipid metabolism are closely associated with the disease,the overall profile of lipid and sterol changes that occur in the brain during Alzheimer’s disease remains unclear.In this study,we compared brain tissues extracted from 32-week-old male wild-type mice and 5×FAD transgenic Alzheimer’s disease model mice,which carry mutations in the amyloid precursor protein(APP)and presenilin 1(PS1)genes.Using untargeted lipidomics and sterolomics techniques,we investigated the metabolic profiles of lipids,with a focus on sterols specifically,in three brain regions:cerebellum,hippocampus,and olfactory bulb.Our results revealed significant alterations in various lipids,particularly in the hippocampus and olfactory bulb,suggesting changes in energy levels in these regions.Further pathway analysis indicated notable disruptions in key metabolic processes,particularly those related to fatty acids and cell membrane components.Additionally,we observed decreased expression of 15 genes involved in lipid and sterol regulation.Collectively,these findings provide new insights into how imbalances in lipid and sterol metabolism may contribute to the progression of Alzheimer’s disease,highlighting potential metabolic pathways involved in the development of this debilitating disease.
基金National Research,Development and Innovation Fund of the Ministry of Culture and Innovation under the National Laboratories Program(National Tumor Biology Laboratory,Grant/Award Number:2022-2.1.1-NL-2022-00010Senior Research Fellowship from National Health and Medical Research Council of Australia,Grant/Award Number:1156693+1 种基金Hungarian Thematic Excellence Program,Grant/Award Number:TKP2021-EGA-44Tour de Cure,Pioneering Grant,Grant/Award Number:RSP-253-18/19。
文摘Realistic models for cancer research representing disease progression that commensurately respond to therapeutics consistent with clinical observation are the holy grail for pre-clinical research and screening.Although such an ideal is elusive,well-characterized in vivo models facilitate our understanding of disease,progression,and therapeutic opportunities.Here,we characterize a commonly used syngeneic BALB/c mouse model of triple negative breast cancer(4T1)after establishing tumors in their flanks.Tumors developed at the subcutaneous injection site for all experimental mice and their volumes were monitored.We quantified a rare subset of breast cancer stemlike cells(CSCs),classified as CD44^(+)/CD24^(−)phenotypes in in vitro and ex vivo cell populations.Chromosome numbers in ex vivo metaphase cells were greater than cells cultured in vitro(89.4±3.4,range of 70-132 and 82.6±1.1,range of 70-128;respectively).Further,we observed different types of chromosome aberrations,including gap,deletion,exchange,interstitial deletion,terminal deletion,ring,dicentric,and Robertsonian translocations.For both sources of cells,the number of aberrations was dominated by deletions,terminal deletions,and Robertsonian translocations.Ex vivo cells exhibited greater prevalence of deletions and terminal deletions,whereas in vitro cells displayed more ring aberrations and Robertsonian translocations.In conclusion,we successfully characterized cancer cells from a syngeneic mouse model of breast cancer in terms of rare CSC proportion and a variety of chromosomal aberrations,which is useful for understanding tumor traits associated with cancer development and therapeutic action.The data act as a valuable resource for other studies using the 4T1 BALB/c model.
基金financial support from the National Natural Science Foundation of China (Nos.82473887 and 21927808)the Scientific and Technological Innovation Program of Shanghai (No.23DZ2202500)the CAMS Innovation Fund for Medical Sciences (No.2021-1-I2M-026)。
文摘The brain's functions are governed by molecular metabolic networks.However,due to the sophisticated spatial organization and diverse activities of the brain,characterizing both the minute and large-scale metabolic activity across the entire brain and its numerous micro-regions remains incredibly challenging.Here,we offer a high-definition spatially resolved metabolomics technique to better understand the metabolic specialization and interconnection throughout the mouse brain using improved ambient mass spectrometry imaging.This method allows for the simultaneous mapping of thousands of metabolites at a 30 μm spatial resolution across the mouse brain,ranging from structural lipids to functional neurotransmitters.This approach effectively reveals the distribution patterns of delicate microregions and their distinctive metabolic characteristics.Using an integrated database,we annotated 259 metabolites,demonstrating that the metabolome and metabolic pathways are unique to each brain microregion.The distribution of metabolites,closely linked to functionally connected brain regions and their interactions,offers profound insights into the complexity of chemical processes and their roles in brain function.An initial dataset for future metabolomics research might be obtained from the high-definition mouse brain's spatial metabolome atlas.
基金Guangdong Province Medical Research Fund Project,Grant/Award Number:B2024112The Scientific Research Special Project of the Joint Construction Project of High-level Hospitals between Guangzhou University of Chinese Medicine and the Scientific Research Fund Project,Grant/Award Number:GZYZS2024G09+2 种基金Special Project of the Research Platform of Guangdong Provincial Department of Traditional Chinese Medicine,Grant/Award Number:20254040the Project of the Incubation Program for the Science and Technology Development of Chinese Medicine Guangdong Laboratory/Hengqin Laboratory,Grant/Award Number:HQL2024PZ043Guangdong Province Natural Science Foundation-Guangzhou-South China Joint Youth Fund Project,Grant/Award Number:2023A1515110849。
文摘Background:In recent decades,the global incidence of dengue fever has been stead-ily increasing,with continuous geographical expansion.Researchers have successfully modeled most clinical symptoms of human dengue fever using interferon type I(IFN-I)or combined IFN-I/II receptor knockout mice infected with dengue virus(DENV).However,this model requires further optimization to better support related studies.Methods:This study aimed to establish a stable dengue infection model by evaluating the effects of different genetic backgrounds and injection routes on DENV infection in interferon receptor knockout mice.We first infected various strains of interferon receptor-deficient mice with DENV and compared their susceptibility based on clini-cal symptoms,viremia levels,organ indices,histopathological findings,and vascular leakage markers.Subsequently,we selected the most susceptible strain to further investigate the impact of different injection methods on infection outcomes.Results:We found that BALB/c background mice with type 1 interferon recep-tor knockout(IFNAR)had the most obvious symptoms.Subsequently,we selected IFNAR−/−BALB/c mice to further explore the effects of different injection methods on dengue virus infection.The results showed that the intraperitoneal injection group had the most severe clinical symptoms,the longest duration of viremia,and the most obvious degree of organ damage.Conclusion:Through systematic screening and optimization,we established a robust animal model of dengue virus infection via intraperitoneal injection in IFNAR−/−BALB/c mice.This model offers a valuable tool for future dengue research.
基金The Korea Centers for Disease Control and Prevention,Grant/Award Number:2022-ER1701-00,2022-NI-041-02,2024-ER1702-00 and 2025-NI-014-00。
文摘Background:Humanized mouse models are essential for studying the human immune response and antibody development.However,conventional models show limited B cell maturation and antigen-specific humoral responses.To overcome these limitations,we used the NOG-EXL mice expressing human interleukin 3(IL-3)and granulocyte-macrophage colony-stimulating factor(GM-CSF)to enhance myeloid and B-cell lineage differentiation.Methods:Human CD34^(+)hematopoietic stem cells(HSC)were transplanted into NOG-EXL mice to produce humanized immune systems.After immune cell reconstitution was confirmed across 12 weeks,the mice were immunized twice with inactivated severe fever with thrombocytopenia syndrome virus(SFTSV)antigens.Peripheral blood mononuclear cells and splenocytes were analyzed using multicolor flow cytometry to assess human immune cell subsets.Antigen-specific immunoglobulin G(IgG)production was quantified using enzyme-linked immunosorbent assay(ELISA),and virus-specific B cells were isolated using antigen-labeled recombinant protein probes.Results:Twelve weeks after transplantation of HSCs into NOG-EXL mice,they exhibited robust engraftment of human leukocytes,including T,B,and dendritic cells,compared to NOG mice.Unlike NOG mice,humanized NOG-EXL mice exhibited an increase in human IgG levels,indicating the production of human antibody responses to antigens.Humanized NOG-EXL mice were immunized twice every 2 weeks with inactivated SFTSV,and antigen-specific human antibodies against the virus were detected in the mouse sera by ELISA.Sera from SFTSV-immunized humanized mice demonstrated neutralizing activity against SFTSV,confirming the induction of functional virus-specific neutralizing antibodies.Antigen-binding IgG-positive human B cells were isolated from mouse splenocytes using recombinant protein probes.Conclusion:This model provides a valuable platform for evaluating humoral immunity and isolating B cells using high-affinity human monoclonal antibodies without genetic engineering.
基金financially supported by grants from the Chang-Gung Memorial Hospital(CMRPG3J0971~3,CMRPVVP0111,and CMRPVVQ0041 to CEWCMRPG3P0101 to HJS)the National Science and Technology Council(113-2628-B-182-001-MY3 and 113-2811-B-182-024 to CEW).
文摘Background:KIT proto-oncogene,receptor tyrosine kinase(KIT,CD117)and platelet-derived growth factor-alpha(PDGFRA)are key drivers of gastrointestinal stromal tumors(GIST),but resistance to targeted therapy often arises from tumor protein p53(p53)alterations and loss of cell cycle control.However,the role of p53 status in GIST therapeutic potential has rarely been studied,so this study aimed to employ both wild-type andmutant p53 GIST models to investigate how p53 dysfunction influences the efficacy of p53 pathway-targeted therapies.Methods:The efficacy of the mouse double minute 2 homolog(MDM2)inhibitor(HDM201)and the Wee1 G2 checkpoint kinase(Wee1)inhibitor(adavosertib)was confirmed in both p53 wild-type(p53 WT)and p53 mutant(p53 MT)GIST cells.The anti-proliferative effects were assessed using the Cell Counting Kit-8(CCK-8)assay.Flow cytometry(FACS)and immunoblotting were employed to evaluate apoptosis and the expression of proteins related to drug efficacy.These findings were further validated in a xenograft model.Results:HDM201 selectively inhibited growth and triggered apoptosis in p53WT GIST cells,while adavosertib was effective mainly in p53 MT cells.Western blot analysis revealed thatHDM201 increased p53 and p21 levels in p53WT cells,and adavosertib affectedWee1 and phospho-cdc2 expression in both p53WT and p53 MT cells.In a xenograft mouse model,HDM201 significantly reduced the tumor volume and weight in p53WTGIST cells,whereas p53MT tumors showed only a moderate size reduction with adavosertib,without significant changes.Conclusions:Our results highlight the importance of p53 status in guiding GIST treatment.p53 WT tumors respond toMDM2 inhibitors,while p53 MTtumors show greater sensitivity toWee1 inhibitors,supporting p53 pathway targeting as a promising strategy for GIST patients.
基金supported by the US National Institutes of Health (R01CA160331, R01CA163377, R01CA202919,R01CA239128, P01AG031862, P50CA228991 to R.G.Z. and K99CA241395 to S.K.)US Department of Defense (OC180109 and OC190181 to R.G.Z.)+2 种基金The Honorable Tina Brozman Foundation for Ovarian Cancer Research (to R.G.Z.)Ovarian Cancer Research Alliance Collaborative Research Development Grant (to R.G.Z.)Core facilities support was provided by a Cancer Centre Support Grant(CA010815) to the Wistar Institute。
文摘Epithelial ovarian cancer(EOC) is the leading cause of gynecological cancer-related mortality in the developed world. EOC is a heterogeneous disease represented by several histological and molecular subtypes. Therefore, exploration of relevant preclinical animal models that consider the heterogenic nature of EOC is of great importance for the development of novel therapeutic strategies that can be translated clinically to combat this devastating disease. In this review, we discuss recent progress in the development of preclinical mouse models for EOC study as well as their advantages and limitations.
基金We thank D.D.Meigs(University of Nebraska Medical Center)and Tonya Cejka(freelance English editor)for editing assistance.C.B.G.is funded by NIH grants R35HG010719,R21GM129559,R21AI143394 and R21DA046831.M.O.is funded by 2016–2017 Tokai University School of Medicine Project Research,the Research Aid from the Institute of Medical Sciences in Tokai University,Grant-in-Aid for Scientific Research(25290035)from MEXTa Grant-in-Aid for Challenging Exploratory Research(15K14371)from JSPS.
文摘Genetically engineered mouse(GEM)models are commonly used in biomedical research.Generating GEMs involve complex set of experimental procedures requiring sophisticated equipment and highly skilled technical staff.Because of these reasons,most research institutes set up centralized core facilities where custom GEMs are created for research groups.Researchers,on the other hand,when they begin thinking about generating GEMs for their research,several questions arise in their minds.For example,what type of model(s)would be best useful for my research,how do I design them,what are the latest technologies and tools available for developing my model(s),and finally how to breed GEMs in my research.As there are several considerations and options in mouse designs,and as it is an expensive and time-consuming endeavor,careful planning upfront can ensure the highest chance of success.In this article,we provide brief answers to several frequently asked questions that arise when researchers begin thinking about generating mouse model(s)for their work.
文摘This paper introduces part of the content in the association standard,T/CAAM0002–2020 Nomenclature and Location of Acupuncture Points for Laboratory Animals Part 3:Mouse.This standard was released by the China Association of Acupuncture and Moxibustion on May 15,2020,implemented on October 31,2020,and published by Standards Press of China.The standard was drafted by the Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,and the Nanjing University of Chinese Medicine.Principal draftsmen:Xiang-hong JING and Xing-bang HUA.Participating draftsmen:Wan-zhu BAI,Bin XU,Dong-sheng XU,Yi GUO,Tie-ming MA,Xin-jun WANG,and Sheng-feng LU.
文摘Background:Over the past few decades,a threefold increase in obesity and type 2 diabetes(T2D)has placed a heavy burden on the health-care system and society.Previous studies have shown correlations between obesity,T2D,and neurodegenera-tive diseases,including dementia.It is imperative to further understand the relation-ship between obesity,T2D,and cognitive deficits.Methods:This investigation tested and evaluated the cognitive impact of obesity and T2D induced by high-fat diet(HFD)and the effect of the host genetic background on the severity of cognitive decline caused by obesity and T2D in collaborative cross(CC)mice.The CC mice are a genetically diverse panel derived from eight inbred strains.Results:Our findings demonstrated significant variations in the recorded phenotypes across different CC lines compared to the reference mouse line,C57BL/6J.CC037 line exhibited a substantial increase in body weight on HFD,whereas line CC005 ex-hibited differing responses based on sex.Glucose tolerance tests revealed significant variations,with some lines like CC005 showing a marked increase in area under the curve(AUC)values on HFD.Organ weights,including brain,spleen,liver,and kidney,varied significantly among the lines and sexes in response to HFD.Behavioral tests using the Morris water maze indicated that cognitive performance was differentially affected by diet and genetic background.Conclusions:Our study establishes a foundation for future quantitative trait loci map-ping using CC lines and identifying genes underlying the comorbidity of Alzheimer's disease(AD),caused by obesity and T2D.The genetic components may offer new tools for early prediction and prevention.
文摘Background:Most mutations in the COL6A3 gene lead to collagen VI-related myopathies.This is due to a reduced expression or mislocalization of the COL6A3 protein.Therefore,studying the consequence of knocking out the Col6a3 gene in mouse models is relevant,but the Col6a3 mouse models reported so far do not entirely abolish COL6A3 protein expression.Methods:Here,we present the development,validation and preliminary phenotypic characterization of a novel CRISPR-based knockout mouse model targeting Col6a3 exon 3(Col6a3^(d3/d3)).Results:In this mouse model,Col6a3 mRNA is still expressed at a similar level to wild-type littermates,although the expected protein is undetectable by mass spectrometry.Histological analysis of Col6a3^(d3/d3)quadriceps revealed an abnormally high frequency of muscle cells with internally nucleated muscle cells,consistent with a myopathy phenotype.Interestingly,Col6a3^(d3/d3)mice are smaller in size,with their fat,muscle,and bone kept proportional compared to wild-type littermates.Conclusions:In summary,we performed the validation and preliminary phenotypic characterization of a novel Col6a3 knockout mouse model that could be further characterized and used to study COL6A3 biology and model collagen VI-associated diseases.
基金supported by the National Natural Science Foundation of China (32471049,32170984,32471188,32200802)Natural Science Foundation of Shandong Province (ZR2023QH110)。
文摘Substantial evidence points to the early onset of peripheral inflammation in the development of Parkinson's disease(PD),supporting the“body-first”hypothesis.However,there remains a notable absence of PD-specific animal models induced by inflammatory cytokines.This study introduces a novel mouse model of PD driven by the proinflammatory cytokine CXCL1,identified in our previous research.The involvement of CXCL1 in PD pathogenesis was validated using subacute and chronic MPTP-induced mouse models.Based on these findings,2-month-old C57BL/6J mice were intravenously administered CXCL1(20 ng/kg/day)for 2 weeks(5 days per week),successfully replicating motor deficits and pathological alterations in the substantia nigra observed in the chronic MPTP model.These results demonstrate the potential of CXCL1-induced inflammation as a mechanism for PD modeling.The model revealed activation of the PPAR signaling pathway in CXCL1-mediated neuronal damage by CXCL1.Linoleic acid,a PPAR-γactivator,significantly mitigated MPTPand CXCL1-induced toxicity and reduced serum CXCL1levels.In addition,the CXCL1-injected mouse model shortened the timeline for developing chronic PD mouse model to 2 weeks,offering an efficient platform for studying inflammation-driven processes in PD.The findings provide critical insights into the inflammatory mechanisms underlying PD and identify promising therapeutic targets for intervention.
基金supported by the National Research Foundation of Korea (NRF)funded by the Ministry of Science,ICT&Future Planning (2022R1A2C2006229,2022R1A6A3A01086868)Korea Dementia Research Project through the Korea Dementia Research Center (KDRC)funded by the Ministry of Health&Welfare and Ministry of Science and ICT,Republic of Korea (RS-2024-00345328)KIST Institutional Grant (2E32851)。
文摘Alzheimer'sdisease(AD)isaprogressive neurodegenerative disorder characterized by cognitive impairment and distinct neuropathological features,including amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis.Developing effective diagnostic,preventative,and therapeutic strategies for AD necessitates the establishment of animal models that accurately recapitulate the pathophysiological processes of the disease.Existing transgenic mouse models have significantly contributed to understanding AD pathology but often fail to replicate the complexity of human AD.Additionally,these models are limited in their ability to elucidate the interplay among amyloid-βplaques,neurofibrillary tangles,and reactive astrogliosis due to the absence of spatially and temporally specific genetic manipulation.In this study,we introduce a novel AD mouse model(APP/PS1-TauP301L-Adeno mice)designed to rapidly induce pathological symptoms and enhance understanding of AD mechanisms.Neurofibrillary tangles and severe reactive astrogliosis were induced by injecting AAVDJ-EF1a-hTauP301L-EGFP and Adeno-GFAP-GFP viruses into the hippocampi of 5-month-old APP/PS1 mice.Three months post-injection,these mice exhibited pronounced astrogliosis,substantial amyloid-βplaque accumulation,extensiveneurofibrillarytangles,accelerated neuronal loss,elevated astrocytic GABA levels,and significant spatial memory deficits.Notably,these pathological features were less severe in AAVTauP301L-expressing APP/PS1 mice without augmented reactive astrogliosis.These findings indicate an exacerbating role of severe reactive astrogliosis in amyloid-βplaque and neurofibrillary tangle-associated pathology.The APP/PS1-TauP301L-Adeno mouse model provides a valuable tool for advancing therapeutic research aimed at mitigating the progression of AD.
基金the European Research Council(ERC Advanced MechAGE-ERC-2016-ADG-741883)the Swiss National Science Foundation(no.188522).
文摘Age-related osteoporosis poses a significant challenge in musculoskeletal health;a condition characterized by reduced bone density and increased fracture susceptibility in older individuals necessitates a better understanding of underlying molecular and cellular mechanisms.Emerging evidence suggests that osteocytes are the pivotal orchestrators of bone remodeling and represent novel therapeutic targets for age-related bone loss.Our study uses the prematurely aged PolgD257A/D257A(PolgA)mouse model to scrutinize age-and sex-related alterations in musculoskeletal health parameters(frailty,grip strength,gait data),bone and particularly the osteocyte lacuno-canalicular network(LCN).Moreover,a new quantitative in silico image analysis pipeline is used to evaluate the alterations in the osteocyte network with aging.Our findings underscore the pronounced degenerative changes in the musculoskeletal health parameters,bone,and osteocyte LCN in PolgA mice as early as 40 weeks,with more prominent alterations evident in aged males.Our findings suggest that the PolgA mouse model serves as a valuable model for studying the cellular mechanisms underlying age-related bone loss,given the comparable aging signs and age-related degeneration of the bone and the osteocyte network observed in naturally aging mice and elderly humans.
基金funded by the budgetary funding to FRC FTM for the project“Post-Genomic High-Tech Research on the Mechanisms of Development of Socially Significant Diseases and Stress-Induced Conditions”(Grant No.125031203556-7).
文摘Background:Glucocorticoids are used as anti-inflammatory drugs for the treatment of various diseases,however,their side effects on normal brain tissue remain underinvestigated.Objectives:The study aimed to investigate dexamethasone(DXM)effects on cell composition and myelin content in the mouse brain tissue.Methods:C57Bl/6 male mice(n 60)received single and ten multiple intraperitoneal DXM injections(2.5 mg/kg),and the studied=parameters were analysed at 1,3,7,10 days after a single DXM injection and 15,30,60,and 90 days after the multiple injections.Oligodendrocytes,microglia,and astrocytes were assayed by immunohistochemistry with specific antibodies(Olig2,CD68,and GFAP,respectively)in the corpus callosum of the normal brain tissue.The myelin content was estimated by staining with LuxolFastBlue.The presence of GFAP isoforms was determined by western blotting.Results:DXM administration did not affect oligodendrocytes in the mouse brain but temporarily significantly decreased myelin content(1.2-fold,p 0.0058;1.4-fold,p 0.0001)at 3–15 days time points.At the same time,DXM significantly=<decreased the number of microglial cells(1.5–3.5-fold,p 0.0001)and significantly increased astrocytes(1.8-fold,p<<0.0001).Prolonged administration of DXM resulted in the decrease of the main GFAPα-isoform(50 kDa)and the appearance of shorter GFAP isoforms(30 kDa,42 kDa,44 kDa)similar to that in some neurodegenerative animal models.Conclusion:DXM can modify the cell composition of the normal mouse brain tissue by decreasing microglial cells and increasing astrocytes.Long-term use of DXM results in the inhibition of myelin formation and the appearance of truncated GFAP isoforms,suggesting its ability to induce neurodegeneration-like changes in the normal mouse brain.
基金CAMS Innovation Fund for Medical Sciences,Grant/Award Number:CIFMS,2021-I2M-1-024The Joint Fund for the Department of Science and Technology of Yunnan Province-Kunming Medical University,Grant/Award Number:202201AY070001-007+1 种基金Open Research Fund Project of Yunnan Provincial Key Laboratory of Pharmacology of Natural Medicines,Grant/Award Number:YKLPNP-G2403The Science and Technology Leading Talent Program of Yunnan Province,Grant/Award Number:202405AB350002。
文摘Background:Spinocerebellar ataxia type 2(SCA2)is a neurodegenerative disease marked by significant clinical and genetic heterogeneity,primarily caused by expanded CAG mutations in the ATXN2 gene.The unstable expansion of CAG repeats disrupts the genetic stability of animal models,which is detrimental to disease research.Methods:In this study,we established a mouse model in which CAG repeats do not undergo microsatellite instability(MSI)across generations.A humanized ATXN2 cDNA with four CAA interruptions within 73 CAG expansions was inserted into the Rosa26 locus of C57BL/6J mice.A 23 CAG control mouse model was also generated to verify ATXN2 integration and expression.Results:In our model,the number of CAG repeats remained stable during transmission,with no CAG repeat expansion observed in 64 parent-to-offspring transmissions.Compared with SCA2-Q23 mice,SCA2-Q73 mice exhibited progressive motor impairment,reduced Purkinje cell count and volume(indicative of cell atrophy),and muscle atrophy.These observations in the mice suggest that the behavioral and neuropathological phenotypes may reflect the features of SCA2 patients.RNA-seq analysis of the gastrocnemius muscle in SCA2-Q73 mice showed significant changes in muscle differentiation and development gene expression at 56 weeks,with no significant differences at 16 weeks compared to SCA2-Q23 mice.The expression level of the Myf6 gene significantly changed in the muscles of aged mice.Conclusion:In summary,the establishment of this model not only provides a stable animal model for studying CAG transmission in SCA2 but also indicates that the lack of long-term neural stimulation leads to muscle atrophy.
基金Funding was provided by the following National Institutes of Health grants:F30-DK123989(May Barakat)R01-GM50875(Luisa A.DiPietro)+1 种基金R35-GM139603(Luisa A.DiPietro)R01-AR065941(Terry W.Moore).Additional funding was provided by the University of Illinois Chicago Chancellor's Translational Science Initiative Award.
文摘The Leprdb/db mouse is a common and well-studied model of type II diabetes mel-litus that is often employed in biomedical research.Despite being one of the most commonly used models for the investigation of diabetic wound healing,there are a few specific guidelines for its husbandry,and wound complications such as infection and expansion are common.This study presents a modified animal husbandry ap-proach for the Leprdb/db mouse to reduce the incidence of complications during wound healing experiments.Compared to standard rodent housing protocols,the use of this modified protocol leads to decreased rates of complications among experimental animals across several experiments.The protocol includes increased cage size,de-creased housing density,and more frequent cage replacements.The use of improved husbandry for the Leprdb/db mouse decreases the total number of animals required,minimizes harm during experimentation,and improves the consistency and reproduc-ibility of wound healing studies.
基金supported in part by grants from the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB0490000 to Z.H.)National Key Research and Development Program(2021YFF0702002 to J.L.,2022YFC2303300 to Z.H.,and 2023YFC2305900 to M.W.)+3 种基金“Youth Commando”project(2023QNTJ-02 TO J.L.)Key Project(2024JZZD-02 to Z.H.)of State Key Laboratory of Virology and BiosafetyWuhan Institute of Virology,the National Natural Science Foundation of China(U22A20336 to Z.H.and Y.Z.)Wuhan Natural Science Foundation(202404071010067 to M.W.and 202404071010068 to J.L.).
文摘Dear Editor,Crimean–Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a severe tick-borne illness with a wide geographical distribution,posing a significant threat with case fatality rates ranging from 5%to 70%(Hawman and Feldmann,2023).Due to the lack of approved vaccines and therapeutics,the World Health Organization(WHO)has listed CCHF as one of the priority diseases(Semper et al.,2024).CCHF initially presents as a nonspecific febrile illness,characterized by fever,malaise,myalgia,and nausea,which can rapidly progress to hemorrhagic disease.The hemorrhagic stage is particularly pronounced in severe cases,with rapid progression to disseminated intravascular coagulation(DIC),overt bleeding,kidney or liver failure,and shock(Frank et al.,2024).Up to date,there is an absence of a suitable animal model that can accurately mimic the coagulopathy and bleeding associated with CCHFV infection.Consequently,our understanding of the pathogenic mechanisms underlying these conditions remains limited(Rodriguez et al.,2022).
基金funding from the Innovation Fund Denmark(grant number 2040-00034B).
文摘Background:The current understanding of diabetic kidney disease(DKD)has significant gaps regarding the underlying pathogenesis.In this study,we aimed to characterize the temporal progression of DKD using a state-of-the-art mouse model of hypertension-accelerated disease,integrating kidney biomarker analysis,histopathology,and glomerular transcriptomic profiling.Methods:Female diabetic db/db mice received a single intravenous dose of adenoassociated virus-mediated renin overexpression(ReninAAV,week 5)and underwent uninephrectomy(UNx,week 4).db/db UNx-ReninAAV mice were terminated at weeks 1,4,8,and 12(n=7–8 per group).Female db/m mice were used as healthy controls.Study endpoints included plasma and urine biochemistry,glomerulosclerosis scoring,quantitative kidney histology,and RNA sequencing of glomeruli isolated using lasercapture microdissection.Results:db/db UNx-ReninAAV mice developed progressive albuminuria(from week 4)and glomerulosclerosis(from week 8).A pathway analysis of clustered gene regulations revealed broad glomerular transcriptome perturbations with signatures of increased extracellular matrix(ECM)turnover from week 8 and early onset of metabolic dysfunction.Markers of glomerular cell types and injury exhibited temporal regulation over the course of DKD,with early and sustained downregulation of endothelial markers,heterogeneous regulation of podocyte markers,and significant mesangial and parietal epithelial aberrations.Furthermore,the upregulation of cell injury markers confirmed progressive glomerular injury in the model.Conclusion:The db/db UNx-ReninAAV mouse model exhibits distinct temporal dynamics in glomerular cell markers,metabolic dysregulation,ECM remodeling,and injury.Together,these results highlight the utility of the db/db UNx-ReninAAV model as a relevant preclinical platform for studying progressive DKD.
