Otoancorin(OTOA)is a glycosylphosphatidylinositol(GPI)-anchored protein mediating the attachment of the tectorial membrane(TM)to the spiral limbus(SL)in the inner ear.Homozygous or compound heterozygous mutations in O...Otoancorin(OTOA)is a glycosylphosphatidylinositol(GPI)-anchored protein mediating the attachment of the tectorial membrane(TM)to the spiral limbus(SL)in the inner ear.Homozygous or compound heterozygous mutations in OTOA cause autosomal recessive deafness(DFNB22).We performed short-read exome sequencing(SRS)in a 10-monthold boy with sensorineural hearing loss,identifying a potential p.Glu787*variant in OTOA.Interestingly,this variant is common among normal-hearing individuals,leading us to question its pathogenic potential.展开更多
Background Megalencephalic leukoencephalopathy with subcortical cysts(MLC)is a rare neurological degenerative disorder caused by the mutations of MLC1 or GLIALCAM with autosomal recessive or autosomal dominant inherit...Background Megalencephalic leukoencephalopathy with subcortical cysts(MLC)is a rare neurological degenerative disorder caused by the mutations of MLC1 or GLIALCAM with autosomal recessive or autosomal dominant inheritance and a different prognosis,characterized by macrocephaly,delayed motor and cognitive development,and bilateral abnormal signals in cerebral white matter(WM)with or without cysts on magnetic resonance imaging(MRI).This study aimed to reveal the clinical and genetic features of MLC patients with GLIALCAM mutations and to explore the brain pathological characteristics and prognosis of mouse models with different modes of inheritance.Methods Clinical information and peripheral venous blood were collected from six families.Genetic analysis was performed by Sanger sequencing of GLIALCAM.Glialcam^(Arg92Trp/+)and Glialcam^(Lys68Met/Thr132Asn)mouse models were generated based on mutations from patients(c.274C>T(p.Arg92Trp)(c.203A>T(p.Lys68Met),and c.395C>A(p.Thr132Asn))).Brain pathologies of the mouse models at different time points were analyzed.Results Six patients were clinically diagnosed with MLC.Of the six patients,five(Pt1-Pt5)presented with a heterozygous mutation in GLIALCAM(c.274C>T(p.Arg92Trp)or c.275G>C(p.Arg92Pro))and were diagnosed with MLC2B;the remaining patient(Pt6)with two compound heterozygous mutations in GLIALCAM(c.203A>T(p.Lys68Met)and c.395C>A(p.Thr132Asn))was diagnosed with MLC2A.The mutation c.275C>G(p.Arg92Pro)has not been reported before.Clinical manifestations of the patient with MLC2A(Pt6)progressed with regression,whereas the course of the five MLC2B patients remained stable or improved.The Glialcam^(Arg92Trp/+)and Glialcam^(Lys68Met/Thr132Asn)mouse models showed vacuolization in the anterior commissural WM at 1 month of age and vacuolization in the cerebellar WM at 3 and 6 months,respectively.At 9 months,the vacuolization of the GlialcamiLys68Met/Thr132Asn mouse model was heavier than that of the Glialcam^(Arg92Trp/+)mouse model.Decreased expression of Glialcam in Glialcam^(Arg92Trp/+)and Glialcam^(Lys68Met/Thr132Asn)mice may contribute to the vacuolization.Conclusions Clinical and genetic characterization of patients with MLC and GLIALCAM mutations revealed a novel mutation,expanding the spectrum of GLIALCAM mutations.The first Glialcam mouse model with autosomal recessive inheritance and a new Glialcam mouse model with autosomal dominant inheritance were generated.The two mouse models with different modes of inheritance showed different degrees of brain pathological features,which were consistent with the patients'phenotype and further confirmed the pathogenicity of the corresponding mutations.展开更多
Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) are one of the most frequent genetic causes of both familial and sporadic Parkinson’s disease (PD). Mounting evidence has demonstrated pathological similarit...Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) are one of the most frequent genetic causes of both familial and sporadic Parkinson’s disease (PD). Mounting evidence has demonstrated pathological similarities between LRRK2-associated PD (LRRK2-PD) and sporadic PD, suggesting that LRRK2 is a potential disease modulator and a thera-peutic target in PD. LRRK2 mutant knock-in (KI) mouse models display subtle alterations in pathological aspects that mirror early-stage PD, including increased susceptibility of nigrostriatal neurotransmission, development of motor and non-motor symptoms, mitochondrial and autophagy-lysosomal defects and synucleinopathies. This review provides a rationale for the use of LRRK2 KI mice to investigate the LRRK2-mediated pathogenesis of PD and implications from current findings from different LRRK2 KI mouse models, and ultimately discusses the therapeutic potentials against LRRK2-associated pathologies in PD.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
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
Huntington＇s disease （HD） is an inherited autosomal dominant neurodegenerative disease characterized by pro- gressive motor deficits, cognitive decline, and psychiatric symptoms. It is caused by a pathological expa...Huntington＇s disease （HD） is an inherited autosomal dominant neurodegenerative disease characterized by pro- gressive motor deficits, cognitive decline, and psychiatric symptoms. It is caused by a pathological expansion of CAG trinucleotide repeats in exon 1 of the HD gene, resulting in the translation of a mutant form of huntingtin protein （mutant Htt） with an expanded polyglutamine domain in the N-terminal region [1 ]. Despite great progress in understanding the pathogenesis of HD using multiple mouse models, the exact mechanisms by which mutant Htt induces neuronal dysfunction and death are still not completely clear, and there is no curative treatment for this disease. An important reason is that the mouse, which is the most widely used animal model in HD research, differs from the human in many aspects, including the physiology, drug metabolism, blood-brain barrier, life span, brain volume, and neuroanatomical organization [2]. Thus, it is necessary to establish HD models with higher species than rodents, such as the dog, pig, and non- human primate, so as to bridge the gap between preclinical mouse models and clinical studies.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
Air mouse has a wide range of uses in robotics,automation,and VR/AR technologies.In this work,the air mouse is prepared using triboelectric sensors,controller units,and machine learning.The triboelectric nanogenerator...Air mouse has a wide range of uses in robotics,automation,and VR/AR technologies.In this work,the air mouse is prepared using triboelectric sensors,controller units,and machine learning.The triboelectric nanogenerator(TENG)performance was optimized by altering the filler’s properties.A dual-ferroelectric crystal system BNKT(xBi_(0.5) Na_(0.5) TiO_(3)-(1−x)Bi_(0.5) K_(0.5) TiO_(3))was prepared with different concentrations(x=_(0.5),0.6,0.7,0.8,and 0.9)to alter the dielectric property.The BNKT-8-based TENG showed a higher performance of 134.04 V and 1.49μA.The prepared device enables to power the small electronic devices such as hygrometers and calculators.Using this TENG device air mouse system with machine learning allows the user to control the mouse pointer in the computer using the smart glove with a high accuracy of 100%.展开更多
The advancement of tissue clearing technology has significantly propelled neuroscience research.Nevertheless,the fluorescent proteins used in traditional transgenic mouse strains were not specifically optimized for ti...The advancement of tissue clearing technology has significantly propelled neuroscience research.Nevertheless,the fluorescent proteins used in traditional transgenic mouse strains were not specifically optimized for tissue clearing procedures,resulting in a substantial decrease in fluorescent intensity after clearing.In this study,we developed the Ci1 reporter mouse strain(where Ci stands for the Chinese Institute for Brain Research,CIBR)based on the bright red fluorescent protein mScarlet.The Ci1 reporter exhibits no fluorescence leakage in various organs or tissue types and can be readily crossed with multiple tissue-specific Cre lines.Compared to the Ai14 mouse strain,the Ci1 reporter strain demonstrates lower non-specific leakage,stronger fluorescence intensity in different tissues,and better preservation of fluorescence following tissue clearing treatment.The creation of the Ci1 reporter provides a more effective tool for both neuroscience and other biomedical research applications.展开更多
Murine subarachnoid hemorrhage(SAH)induced using the filament perforation method is a useful in vivo experimental model to investigate the pathophysiological mechanisms in the brain underlying SAH.However,identifying ...Murine subarachnoid hemorrhage(SAH)induced using the filament perforation method is a useful in vivo experimental model to investigate the pathophysiological mechanisms in the brain underlying SAH.However,identifying mice with comorbid acute neurogenic pulmonary edema(NPE),a life-threatening systemic consequence often induced by SAH,in this model is difficult without histopathological investiga-tions.Herein,we present an imaging procedure involving dual-energy X-ray absorp-tiometry(DXA)to identify NPE in a murine model of SAH.We quantified the lung lean mass(LM)and compared the relationship between micro-computed tomography(CT)evidence of Hounsfield unit(HU)values and histopathological findings of PE.Of the 85 mice with successful induction of SAH by filament perforation,16(19%)had NPE,as verified by postmortem histology.The DXA-LM values correlate well with CT-HU levels(r=0.63,p<0.0001).Regarding the relationship between LM and HU in mice with post-SAH NPE,the LM was positively associated with HU values(r2=0.43;p=0.0056).A receiver operating characteristics curve of LM revealed a sensitivity of 87%and specificity of 57%for detecting PE,with a similar area under the curve as the HU(0.79±0.06 vs.0.84±0.07;p=0.21).These data suggest that confirming acute NPE using DXA-LM is a valuable method for selecting a clinically relevant murine NPE model that could be used in future experimental SAH studies.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
Ostreopsis cf.ovata is a marine benthic dinoflagellate in tropical and temperate seas and can produce potent toxic compounds.The existence of O.cf.ovata has been found in the Chinese coastal areas,but studies on its t...Ostreopsis cf.ovata is a marine benthic dinoflagellate in tropical and temperate seas and can produce potent toxic compounds.The existence of O.cf.ovata has been found in the Chinese coastal areas,but studies on its toxicity are very few.This study investigated the toxicity of the O.cf.ovata(TIO991)isolated from Weizhou Island in the South China Sea by using methanol and chloroform to extract toxic compounds from the algal cells cultured indoor.Experiments on mouse acute toxicity showed that the crude methanol extract(CME)of O.cf.ovata caused the death of mice in 16–18 min.Furthermore,CME inhibited the cell reproduction of human neuroblastoma cells(BE(2)-M17 cells)by Cell Counting Kit-8 with a dose-and time-effect relationship and caused cell death in the form of cell necrosis.We found that CME had strong hemolytic activity and was significantly inhibited by ouabain,indicating that CME might contain palytoxins.By contrast,the crude chloroform extract of O.cf.ovata was relatively weak in toxicity as obtained in our experiments on mouse acute toxicity,cytotoxicity,and hemolytic activity.This suggests that the algae may raise the potential threat to marine ecosystems and public health.展开更多
Background:Rabies virus(RABV)-derived neuronal tracing tools are extensively applied in retrograde tracing due to their strict retrograde transsynaptic transfer property and low neurotoxicity.However,the RABV infectio...Background:Rabies virus(RABV)-derived neuronal tracing tools are extensively applied in retrograde tracing due to their strict retrograde transsynaptic transfer property and low neurotoxicity.However,the RABV infection and expression of fluorescence products would be gradually cleared while the infected neurons still survive,a phenomenon known as non-cytolytic immune clearance(NCLIC).This phenomenon introduced the risk of fluorescence loss and led to the omission of a subset of neurons that should be labeled,thereby interfering in the analysis of tracing results.Methods:To compensate for the fluorescence loss problem,in this study,we developed a novel marker footprints(MF)mouse,involving a Cre recombinase-dependent red fluorescent reporter system and systemic expression of glycoprotein(G)and ASLV-A receptor(TVA).Using this mouse model combined with the well-developed RABV-EnvA-ΔG-GFP-Cre viral tool,we developed a novel green-to-red spectral labeling strategy.Results:Neurons in the MF mouse could be co-labeled with green fluorescence from the very quick expression of the viral tool and with red fluorescence from the relatively slow expression of the neuron itself,so neurons undergoing NCLIC with green fluorescence loss could be relabeled red.Furthermore,newly infected neurons could be labeled green and other neurons could be labeled yellow due to the temporal expression difference between the two fluorescent proteins.Conclusions:This is the first polysynaptic retrograde tracing labeling strategy that could label neurons using spectral fluorescence colors with only one injection of the viral tool,enabling its application in recognizing the labeling sequence of neurons in brain regions and enhancing the spatiotemporal resolution of neuronal tracing.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
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
Melanosomes are specialized membrane-bound organelles within which melanin is synthesized and stored.The levels of melanin can be effectively reduced by inhibiting melanin synthesis or promoting melanosome degradation...Melanosomes are specialized membrane-bound organelles within which melanin is synthesized and stored.The levels of melanin can be effectively reduced by inhibiting melanin synthesis or promoting melanosome degradation via autophagy.Ceramide,a central molecule in sphingolipid metabolism,has been widely implicated in the regulation of autophagy.Few researchers have addressed the potential effects of ceramide analogs on suppressing melanin synthesis.However,whether ceramide can induce melanosome autophagy and the potential autophagy-dependent mechanism underlying this phenomenon remain unknown.Here,an active compound from the marine microalgae Emiliania huxleyi extract was firstly isolated and identified as a long-chain C22-ceramide(C22-Cer).In vitro results of mouse B16 melanoma cell experiments showed that treatment with 2-5µmol/L C22-Cer significantly suppressed the increase ofα-MSH-induced melanin levels and tyrosinase activity without cytotoxicity.C22-Cer induced typical hallmarks of autophagy such as accumulation of autophagosomes,enhanced autophagic flux and microtubule-associated protein light chain 3,LC3-II expression,and p62 degradation through activating c-Jun N-terminal kinase(JNK)directly.Furthermore,C22-Cer activated JNK-Bcl-2 signaling,dissociated the Beclin1/Bcl-2 complex,and induced melanosome autophagy without affecting the expression of MITF.Besides,the Ca^(2+)influx induced by treatment with C22-Cer further increased the substantial accumulation of autophagosomes.Together,we found a novel marine-derived compound,C22-Cer,targeting JNK pathway and Ca^(2+)signaling to induce melanosome autophagy and suppress melanin accumulation in B16 cells.This study implicates that C22-Cer might be a potential therapeutic mediator against skin pigmentation in mammals.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
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korean government(MSIT)(No.2021R1C1C1007980 to B.J.K.)Chungnam National University Sejong Hospital Research Fund,2022,and Chungnam National University(to B.J.K.)+6 种基金supported by the Basic Science Research Program through the NRF,funded by the Ministry of Education(No.2021R1A2C2092038 to B.Y.C.)Bio Core Facility Center program(No.NRF-2022M3A9G1014007 to B.Y.C.)the Basic Research Laboratory program through the NRF,funded by the Ministry of Education(No.RS-2023-0021971031482092640001 to B.Y.C.)the Technology Innovation Program(No.K_G012002572001 to B.Y.C.)funded By the Ministry of Trade,Industry&Energy(MOTIE,Korea)funded by SNUBH(Seoul National University Bundang Hospital)intramural research fund(No.13-2022-0010,02-2017-0060,16-2023-0002,13-2023-0002,16-2022-0005,13-2024-0004,and 13-2017-0013 to B.Y.C.)supported by the National Institute on Deafness and Other Communication Disorders(NIDCD)part of the US National Institutes of Health(No.R01DC018814 to S.P.).
文摘Otoancorin(OTOA)is a glycosylphosphatidylinositol(GPI)-anchored protein mediating the attachment of the tectorial membrane(TM)to the spiral limbus(SL)in the inner ear.Homozygous or compound heterozygous mutations in OTOA cause autosomal recessive deafness(DFNB22).We performed short-read exome sequencing(SRS)in a 10-monthold boy with sensorineural hearing loss,identifying a potential p.Glu787*variant in OTOA.Interestingly,this variant is common among normal-hearing individuals,leading us to question its pathogenic potential.
基金funded by the National Natural Science Foundation of China(Grant Number:81741053,81501123)the Beijing Natural Science Foundation(Grant Number:7151010,7172217)+5 种基金the Bejing Municipal Science&Technology Commission(Grant Number:Z161100000216133,Z161100004916169)the Beijing Institute for Brain Disorders Foundation(Grant Number:BIBDPXM2014_014226_000016)the Beijing Municipal Natural Science Key Project(Grant Number 15G10050)Bejing key laboratory of molecular diagnosis and study on pediatric genetic discases(Grant Number BZ0317)the National Key Rescarch and Development Program of China(Grant Number:2016YFC1306201,2016YFC0901505)the Fundamental Research Funds for the Central Universities(Grant Number:BMU2017JI002).
文摘Background Megalencephalic leukoencephalopathy with subcortical cysts(MLC)is a rare neurological degenerative disorder caused by the mutations of MLC1 or GLIALCAM with autosomal recessive or autosomal dominant inheritance and a different prognosis,characterized by macrocephaly,delayed motor and cognitive development,and bilateral abnormal signals in cerebral white matter(WM)with or without cysts on magnetic resonance imaging(MRI).This study aimed to reveal the clinical and genetic features of MLC patients with GLIALCAM mutations and to explore the brain pathological characteristics and prognosis of mouse models with different modes of inheritance.Methods Clinical information and peripheral venous blood were collected from six families.Genetic analysis was performed by Sanger sequencing of GLIALCAM.Glialcam^(Arg92Trp/+)and Glialcam^(Lys68Met/Thr132Asn)mouse models were generated based on mutations from patients(c.274C>T(p.Arg92Trp)(c.203A>T(p.Lys68Met),and c.395C>A(p.Thr132Asn))).Brain pathologies of the mouse models at different time points were analyzed.Results Six patients were clinically diagnosed with MLC.Of the six patients,five(Pt1-Pt5)presented with a heterozygous mutation in GLIALCAM(c.274C>T(p.Arg92Trp)or c.275G>C(p.Arg92Pro))and were diagnosed with MLC2B;the remaining patient(Pt6)with two compound heterozygous mutations in GLIALCAM(c.203A>T(p.Lys68Met)and c.395C>A(p.Thr132Asn))was diagnosed with MLC2A.The mutation c.275C>G(p.Arg92Pro)has not been reported before.Clinical manifestations of the patient with MLC2A(Pt6)progressed with regression,whereas the course of the five MLC2B patients remained stable or improved.The Glialcam^(Arg92Trp/+)and Glialcam^(Lys68Met/Thr132Asn)mouse models showed vacuolization in the anterior commissural WM at 1 month of age and vacuolization in the cerebellar WM at 3 and 6 months,respectively.At 9 months,the vacuolization of the GlialcamiLys68Met/Thr132Asn mouse model was heavier than that of the Glialcam^(Arg92Trp/+)mouse model.Decreased expression of Glialcam in Glialcam^(Arg92Trp/+)and Glialcam^(Lys68Met/Thr132Asn)mice may contribute to the vacuolization.Conclusions Clinical and genetic characterization of patients with MLC and GLIALCAM mutations revealed a novel mutation,expanding the spectrum of GLIALCAM mutations.The first Glialcam mouse model with autosomal recessive inheritance and a new Glialcam mouse model with autosomal dominant inheritance were generated.The two mouse models with different modes of inheritance showed different degrees of brain pathological features,which were consistent with the patients'phenotype and further confirmed the pathogenicity of the corresponding mutations.
基金Tai Hung Fai Charitable Foundation-Edwin S H Leong Research Programme for Parkinson’s DiseaseThe Henry G.Leong Endowed Professorship in Neurology+1 种基金The Donation Fund for Neurology ResearchHealth and Medical Research Fund(HMRF),Food and Health Bureau,Hong Kong S.A.R.
文摘Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) are one of the most frequent genetic causes of both familial and sporadic Parkinson’s disease (PD). Mounting evidence has demonstrated pathological similarities between LRRK2-associated PD (LRRK2-PD) and sporadic PD, suggesting that LRRK2 is a potential disease modulator and a thera-peutic target in PD. LRRK2 mutant knock-in (KI) mouse models display subtle alterations in pathological aspects that mirror early-stage PD, including increased susceptibility of nigrostriatal neurotransmission, development of motor and non-motor symptoms, mitochondrial and autophagy-lysosomal defects and synucleinopathies. This review provides a rationale for the use of LRRK2 KI mice to investigate the LRRK2-mediated pathogenesis of PD and implications from current findings from different LRRK2 KI mouse models, and ultimately discusses the therapeutic potentials against LRRK2-associated pathologies in PD.
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.
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.
文摘Huntington＇s disease （HD） is an inherited autosomal dominant neurodegenerative disease characterized by pro- gressive motor deficits, cognitive decline, and psychiatric symptoms. It is caused by a pathological expansion of CAG trinucleotide repeats in exon 1 of the HD gene, resulting in the translation of a mutant form of huntingtin protein （mutant Htt） with an expanded polyglutamine domain in the N-terminal region [1 ]. Despite great progress in understanding the pathogenesis of HD using multiple mouse models, the exact mechanisms by which mutant Htt induces neuronal dysfunction and death are still not completely clear, and there is no curative treatment for this disease. An important reason is that the mouse, which is the most widely used animal model in HD research, differs from the human in many aspects, including the physiology, drug metabolism, blood-brain barrier, life span, brain volume, and neuroanatomical organization [2]. Thus, it is necessary to establish HD models with higher species than rodents, such as the dog, pig, and non- human primate, so as to bridge the gap between preclinical mouse models and clinical studies.
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financially supported by the Basic Science Re-search Program(No.RS-2023-NR077252)Regional Leading Re-search Center(No.RS-2024-00405278)through the National Re-search Foundation of Korea(NRF)grant funded by the Korea Gov-ernment(MSIT).
文摘Air mouse has a wide range of uses in robotics,automation,and VR/AR technologies.In this work,the air mouse is prepared using triboelectric sensors,controller units,and machine learning.The triboelectric nanogenerator(TENG)performance was optimized by altering the filler’s properties.A dual-ferroelectric crystal system BNKT(xBi_(0.5) Na_(0.5) TiO_(3)-(1−x)Bi_(0.5) K_(0.5) TiO_(3))was prepared with different concentrations(x=_(0.5),0.6,0.7,0.8,and 0.9)to alter the dielectric property.The BNKT-8-based TENG showed a higher performance of 134.04 V and 1.49μA.The prepared device enables to power the small electronic devices such as hygrometers and calculators.Using this TENG device air mouse system with machine learning allows the user to control the mouse pointer in the computer using the smart glove with a high accuracy of 100%.
基金supported by the startup funding from the Chinese Institute for Brain Research to Hu Zhao.
文摘The advancement of tissue clearing technology has significantly propelled neuroscience research.Nevertheless,the fluorescent proteins used in traditional transgenic mouse strains were not specifically optimized for tissue clearing procedures,resulting in a substantial decrease in fluorescent intensity after clearing.In this study,we developed the Ci1 reporter mouse strain(where Ci stands for the Chinese Institute for Brain Research,CIBR)based on the bright red fluorescent protein mScarlet.The Ci1 reporter exhibits no fluorescence leakage in various organs or tissue types and can be readily crossed with multiple tissue-specific Cre lines.Compared to the Ai14 mouse strain,the Ci1 reporter strain demonstrates lower non-specific leakage,stronger fluorescence intensity in different tissues,and better preservation of fluorescence following tissue clearing treatment.The creation of the Ci1 reporter provides a more effective tool for both neuroscience and other biomedical research applications.
基金supported by the Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science KAKENHI 22K09110.
文摘Murine subarachnoid hemorrhage(SAH)induced using the filament perforation method is a useful in vivo experimental model to investigate the pathophysiological mechanisms in the brain underlying SAH.However,identifying mice with comorbid acute neurogenic pulmonary edema(NPE),a life-threatening systemic consequence often induced by SAH,in this model is difficult without histopathological investiga-tions.Herein,we present an imaging procedure involving dual-energy X-ray absorp-tiometry(DXA)to identify NPE in a murine model of SAH.We quantified the lung lean mass(LM)and compared the relationship between micro-computed tomography(CT)evidence of Hounsfield unit(HU)values and histopathological findings of PE.Of the 85 mice with successful induction of SAH by filament perforation,16(19%)had NPE,as verified by postmortem histology.The DXA-LM values correlate well with CT-HU levels(r=0.63,p<0.0001).Regarding the relationship between LM and HU in mice with post-SAH NPE,the LM was positively associated with HU values(r2=0.43;p=0.0056).A receiver operating characteristics curve of LM revealed a sensitivity of 87%and specificity of 57%for detecting PE,with a similar area under the curve as the HU(0.79±0.06 vs.0.84±0.07;p=0.21).These data suggest that confirming acute NPE using DXA-LM is a valuable method for selecting a clinically relevant murine NPE model that could be used in future experimental SAH studies.
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 Special Research for the Science and Technology Basic Resources Investigation Program of China(No.2018FY100200)the National Natural Science Foundation of China(No.42176206)the Natural Science Foundation of Shandong Province(No.ZR2021MD071)。
文摘Ostreopsis cf.ovata is a marine benthic dinoflagellate in tropical and temperate seas and can produce potent toxic compounds.The existence of O.cf.ovata has been found in the Chinese coastal areas,but studies on its toxicity are very few.This study investigated the toxicity of the O.cf.ovata(TIO991)isolated from Weizhou Island in the South China Sea by using methanol and chloroform to extract toxic compounds from the algal cells cultured indoor.Experiments on mouse acute toxicity showed that the crude methanol extract(CME)of O.cf.ovata caused the death of mice in 16–18 min.Furthermore,CME inhibited the cell reproduction of human neuroblastoma cells(BE(2)-M17 cells)by Cell Counting Kit-8 with a dose-and time-effect relationship and caused cell death in the form of cell necrosis.We found that CME had strong hemolytic activity and was significantly inhibited by ouabain,indicating that CME might contain palytoxins.By contrast,the crude chloroform extract of O.cf.ovata was relatively weak in toxicity as obtained in our experiments on mouse acute toxicity,cytotoxicity,and hemolytic activity.This suggests that the algae may raise the potential threat to marine ecosystems and public health.
基金Hubei Natural Science Foundation of China,Grant/Award Number:2024AFB593。
文摘Background:Rabies virus(RABV)-derived neuronal tracing tools are extensively applied in retrograde tracing due to their strict retrograde transsynaptic transfer property and low neurotoxicity.However,the RABV infection and expression of fluorescence products would be gradually cleared while the infected neurons still survive,a phenomenon known as non-cytolytic immune clearance(NCLIC).This phenomenon introduced the risk of fluorescence loss and led to the omission of a subset of neurons that should be labeled,thereby interfering in the analysis of tracing results.Methods:To compensate for the fluorescence loss problem,in this study,we developed a novel marker footprints(MF)mouse,involving a Cre recombinase-dependent red fluorescent reporter system and systemic expression of glycoprotein(G)and ASLV-A receptor(TVA).Using this mouse model combined with the well-developed RABV-EnvA-ΔG-GFP-Cre viral tool,we developed a novel green-to-red spectral labeling strategy.Results:Neurons in the MF mouse could be co-labeled with green fluorescence from the very quick expression of the viral tool and with red fluorescence from the relatively slow expression of the neuron itself,so neurons undergoing NCLIC with green fluorescence loss could be relabeled red.Furthermore,newly infected neurons could be labeled green and other neurons could be labeled yellow due to the temporal expression difference between the two fluorescent proteins.Conclusions:This is the first polysynaptic retrograde tracing labeling strategy that could label neurons using spectral fluorescence colors with only one injection of the viral tool,enabling its application in recognizing the labeling sequence of neurons in brain regions and enhancing the spatiotemporal resolution of neuronal tracing.
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supported by the National Natural Science Foundation of China(Nos.42076086 and 32202068)Fujian Province Natural Science Foundation of China(Nos.2019J01696 and 2022J01332)the Fujian Province Young and Middle-Aged Teacher Education Research Project(No.JAT200247).
文摘Melanosomes are specialized membrane-bound organelles within which melanin is synthesized and stored.The levels of melanin can be effectively reduced by inhibiting melanin synthesis or promoting melanosome degradation via autophagy.Ceramide,a central molecule in sphingolipid metabolism,has been widely implicated in the regulation of autophagy.Few researchers have addressed the potential effects of ceramide analogs on suppressing melanin synthesis.However,whether ceramide can induce melanosome autophagy and the potential autophagy-dependent mechanism underlying this phenomenon remain unknown.Here,an active compound from the marine microalgae Emiliania huxleyi extract was firstly isolated and identified as a long-chain C22-ceramide(C22-Cer).In vitro results of mouse B16 melanoma cell experiments showed that treatment with 2-5µmol/L C22-Cer significantly suppressed the increase ofα-MSH-induced melanin levels and tyrosinase activity without cytotoxicity.C22-Cer induced typical hallmarks of autophagy such as accumulation of autophagosomes,enhanced autophagic flux and microtubule-associated protein light chain 3,LC3-II expression,and p62 degradation through activating c-Jun N-terminal kinase(JNK)directly.Furthermore,C22-Cer activated JNK-Bcl-2 signaling,dissociated the Beclin1/Bcl-2 complex,and induced melanosome autophagy without affecting the expression of MITF.Besides,the Ca^(2+)influx induced by treatment with C22-Cer further increased the substantial accumulation of autophagosomes.Together,we found a novel marine-derived compound,C22-Cer,targeting JNK pathway and Ca^(2+)signaling to induce melanosome autophagy and suppress melanin accumulation in B16 cells.This study implicates that C22-Cer might be a potential therapeutic mediator against skin pigmentation in mammals.
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.
