The proper flowering time of rose(Rosa hybrida)is vital for the market value of this horticultural crop,but the mechanism regulating this trait is largely unclear.Here,we found that the transcription factor SQUAMOSA P...The proper flowering time of rose(Rosa hybrida)is vital for the market value of this horticultural crop,but the mechanism regulating this trait is largely unclear.Here,we found that the transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE4(RhSPL4)positively regulates flowering time in rose.Transient silencing or overexpression transgenic rose plants of RhSPL4 exhibited delayed or early flowering,respectively.Analysis of transcriptome data from transgenic lines overexpressing RhSPL4 compared to the wild type indicated that differentially expressed genes were significantly enriched in the circadian rhythm pathway.Among the proteins encoded by these genes,RhSPL4 binds to the promoter of PSEUDO-RESPONSE REGULATOR 5-LIKE(RhPRR5L),as revealed in yeast one-hybrid,dual-Luciferase/Renilla luciferase reporter,chromatin immunoprecipitation-quantitative PCR and electrophoretic mobility shift assay.Furthermore,RhSPL4 specifically binds to the478 to441 bp region of the RhPRR5L promoter and activates its transcription.The silencing of RhPRR5L delayed flowering time in rose,resembling the phenotype of RhSPL4-silenced plants.Together,these results indicate that the RhSPL4-RhPRR5L module positively regulates flowering time in rose,laying the foundation for the genetic improvement of flowering time in this important horticultural crop.展开更多
Carotenoid isomerase(CRTISO)is an important enzyme in carotenoid biosynthesis,and it catalyzes the conversion of lycopene precursors to lycopene in several plant species.However,the role of CRTISO in other biochemical...Carotenoid isomerase(CRTISO)is an important enzyme in carotenoid biosynthesis,and it catalyzes the conversion of lycopene precursors to lycopene in several plant species.However,the role of CRTISO in other biochemical processes during plant growth and development remains unclear.Here,we showed that Chinese kale boacrtiso mutants have distinctive characteristics,including a yellowgreen hue and glossy appearance,and this contrasts with the dark green and glaucous traits observed in wild-type(WT)plants.Analysis of pigments in mutants revealed that the reduction in the content of carotenoids and chlorophylls contributed to the yellow-green coloration observed in mutants.An examination of cuticular waxes in Chinese kale indicated that there was a decrease in both the total wax content and the content of individual waxes in boacrtiso mutants(bearing a mutation of BoaCRTISO),which may be caused by the decrease of abscisic acid(ABA)content.The expression of carotenoid,chlorophyll,ABA,and wax biosynthesis genes was downregulated in boacrtiso mutants.This finding confirms that BoaCRTISO regulates the biosynthesis of pigments,ABA,and cuticular waxes in Chinese kale.Our results provide new insights into the interplay between plant pigment and cuticular wax metabolic pathways in Brassica vegetables.展开更多
Red fruit peel is one of pear's most valuable economic traits and is mainly determined by anthocyanins.Many pear cultivars with a red peel originated from bud sports;however,little is known about the genetic mecha...Red fruit peel is one of pear's most valuable economic traits and is mainly determined by anthocyanins.Many pear cultivars with a red peel originated from bud sports;however,little is known about the genetic mechanisms underlying this trait.We have previously identified a mutant Pp BBX24 containing a 14-nucleotide deletion in the coding region(Ppbbx24-del)as the only known variant associated with the red coloration of the mutant‘Red Zaosu'pear(Pyrus pyrifolia White Pear Group).Herein,we analyzed the role of the mutant gene in red coloration and its mechanism of action.The results showed that light promoted red peel coloration in the‘Red Zaosu'pear,and Ppbbx24-del positively affected light-induced anthocyanin biosynthesis,while normal Pp BBX24 had the opposite effects.Transient and stable transformation experiments confirmed that Ppbbx24-del could promote anthocyanin accumulation in pear fruit peels,calli,and tobacco flowers.Due to the loss of nuclear localization sequence(NLS)and viral protein(VP)domains,Ppbbx24-del co-localized in the nucleus and cytoplasm,whereas PpBBX24 localized only in the nucleus.Real-time PCR and transcriptome analyses indicated that PpM YB10 and PpH Y5 are highly expressed in the‘Red Zaosu'pear.In yeast one-hybrid and dual-luciferase assays,Ppbbx24-del and PpHY5 independently promoted the expression of PpC HS,PpC HI,and PpM YB10 by binding to their promoters;however,PpBBX24 did not affect the expression of these genes.Additionally,we found that Ppbbx24-del and PpHY5 had additive effects on the expression of PpC HS,PpC HI,and PpMYB10,as they promote the expression of anthocyanin synthesis genes separately.The co-expression of PpB BX24 and PpHY5 inhibited the activation of downstream genes by PpHY5,which was attributed to the interaction between the two loci.In conclusion,our results clarify the molecular mechanism by which mutant Ppbbx24-del and PpBBX24 exert opposite effects in regulating anthocyanin accumulation in pear.These findings lay an important theoretical foundation for using Ppbbx24-del to create red pear cultivars.展开更多
Coloration in rose(Rosa hybrida) petals is primarily determined by anthocyanin accumulation in vacuoles, and vacuolar acidification plays a central role in controlling the accumulation of this pigment. Nevertheless, t...Coloration in rose(Rosa hybrida) petals is primarily determined by anthocyanin accumulation in vacuoles, and vacuolar acidification plays a central role in controlling the accumulation of this pigment. Nevertheless, the regulatory interplay between anthocyanin accumulation and tissue acidification processes remains somewhat unclear. The present study characterized an activator Rh MYB114 and a repressor Rh MYB16,which functioned synergistically in anthocyanin accumulation and tissue acidification in rose. Transforming tobacco and roses by overexpression, the introduction of Rh MYB114 resulted in an increase in anthocyanin levels and a noticeable decrease in p H in the petal cells of both rose and tobacco, whereas Rh MYB16 introduction led to inverse effects. To further clarify the underlying the regulatory mechanisms, the yeast two-hybrid(Y2H), bimolecular fluorescence complementation(Bi FC) and dual-luciferase(LUC) were employed. The results showed that Rh MYB16 competed with Rh MYB114, bound to Rhb HLH3 or Rhb HLH33, and inhibited its ability to induce the expression of genes related to anthocyanin biosynthesis and acidification. Our findings revealed a feedback mechanism for the regulation of anthocyanin synthesis and tissue acidification involving Rh MYB114, which stimulated the transcriptional expression of Rh MYB16, whose encoded protein Rh MYB16, in turn, negatively regulated the transcriptional expression of Rh MYB114. Therefore, this study underscores the pivotal roles of the Rh MYB114-Rh MYB16 loop in regulating anthocyanin synthesis and tissue acidification, offering insights into metabolic manipulation to enhance the aesthetic appeal of roses.展开更多
Objective Glioma is a highly heterogeneous and malignant intracranial tumor that presents challenges for clinical treatment.ELMO domain containing 2(ELMOD2)is a GTPase-activating protein that regulates a range of cell...Objective Glioma is a highly heterogeneous and malignant intracranial tumor that presents challenges for clinical treatment.ELMO domain containing 2(ELMOD2)is a GTPase-activating protein that regulates a range of cellular biological processes.However,its specific role and prognostic value in tumorigenesis are still unknown.This study aimed to assess the prognostic relevance and signaling function of ELMOD2 in gliomas.Methods The Chinese Glioma Genome Atlas(CGGA)and The Cancer Genome Atlas(TCGA)databases were utilized to conduct a comprehensive analysis of the expression profile of ELMOD2 in gliomas,elucidating its associations with clinicopathological parameters and patient prognosis.Single-cell analysis was performed to characterize ELMOD2 expression across distinct glioma cell subpopulations.Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,and Gene Set Variation Analysis(GSVA)were employed to evaluate the potential biological functions of ELMOD2 in gliomagenesis.Specific small interfering RNAs(siRNAs)were used to knock down ELMOD2 in the glioma cell lines U251 and A172 to assess their cellular behaviors and examine the levels of multiple key signaling molecules associated with the occurrence of gliomas.Results ELMOD2 was overexpressed in gliomas,and this upregulation was correlated with tumor grade,isocitrate dehydrogenase mutation,and 1p/19q codeletion status.Notably,ELMOD2 expression was elevated in classical and mesenchymal subtypes,and single-cell resolution analysis revealed predominant enrichment within malignant cells.Functionally,ELMOD2 regulated cell cycle progression,and its overexpression was related to independent adverse outcomes.In vitro experiments revealed that ELMOD2 was located in the cytoplasm and nucleoplasm.Furthermore,ELMOD2 knockdown reduced proliferation,migration,and invasion and increased apoptosis in U251 and A172 cell lines.Finally,ELMOD2 knockdown significantly decreased p-Erk1/2.Conclusions ELMOD2 expression in glioma is positively correlated with tumorigenesis and is a crucial independent prognostic marker.Thus,ELMOD2 is a promising biomarker and therapeutic target for glioma treatment.展开更多
RANKL signaling is essential for osteoclastogenesis. Its role in osteoblastic differentiation and bone formation is unknown. Here we demonstrate that RANK is expressed at an early stage of bone marrow mesenchymal stem...RANKL signaling is essential for osteoclastogenesis. Its role in osteoblastic differentiation and bone formation is unknown. Here we demonstrate that RANK is expressed at an early stage of bone marrow mesenchymal stem cells(BMSCs) during osteogenic differentiation in both mice and human and decreased rapidly. RANKL signaling inhibits osteogenesis by promoting β-catenin degradation and inhibiting its synthesis. In contrast, RANKL signaling has no significant effects on adipogenesis of BMSCs.Interestingly, conditional knockout of rank in BMSCs with Prx1-Cre mice leads to a higher bone mass and increased trabecular bone formation independent of osteoclasts. In addition, rank: Prx1-Cre mice show resistance to ovariectomy-(OVX) induced bone loss. Thus, our results reveal that RANKL signaling regulates both osteoclasts and osteoblasts by inhibition of osteogenic differentiation of BMSCs and promotion of osteoclastogenesis.展开更多
Age-related osteoporosis is associated with the reduced capacity of bone marrow mesenchymal stem cells （BMSCs） to differentiate into osteoblasts instead of adipocytes. However, the molecular mechanisms that decide t...Age-related osteoporosis is associated with the reduced capacity of bone marrow mesenchymal stem cells （BMSCs） to differentiate into osteoblasts instead of adipocytes. However, the molecular mechanisms that decide the fate of BMSCs remain unclear. In our study, microRNA-23a, and microRNA-23b （miR-23a/b） were found to be markedly downregulated in BMSCs of aged mice and humans. The overexpression of miR-23a/b in BMSCs promoted osteogenic differentiation, whereas the inhibition of miR-23a/b increased adipogenic differentiation. Transmembrane protein 64 （Tmem64）, which has expression levels inversely related to those of miR-23a/b in aged and young mice, was identified as a major target of miR-23a/b during BMSC differentiation. In conclusion, our study suggests that miR-23a/b has a critical role in the regulation of mesenchymal lineage differentiation through the suppression of Tmem64.展开更多
In the recent two decades, it has been well elucidated that receptor activator of nuclear factor-κB ligand （RANKL; also known as TNFSF11） binding to its receptor RANK （also known as TNFRSF11A） drives osteoclast d...In the recent two decades, it has been well elucidated that receptor activator of nuclear factor-κB ligand （RANKL; also known as TNFSF11） binding to its receptor RANK （also known as TNFRSF11A） drives osteoclast development as the crucial signaling pathway.;However, accumulating evidence also implies that展开更多
The essential effect of vitamin A on immune function occurs through various mechanisms including direct effect on ThloTh2 balance modulation. However, it is unclear whether or not vitamin A can regulate Thl-Th2 balanc...The essential effect of vitamin A on immune function occurs through various mechanisms including direct effect on ThloTh2 balance modulation. However, it is unclear whether or not vitamin A can regulate Thl-Th2 balance under a strong Thl-polarizing condition. Therefore, the purpose of our study was to examine the effect of vitamin A metabolite allotrans retinoic acid （ATRA） on ThloTh2 differentiation in CD4~ T cells under GATA-3 deficiency, which can induce Thl-polarizing condition. In the present study, GATA-3 deficiency T cells were induced by siRNA and checked by real-time quantitative PCR and western blot. GATA-3 deficiency CD4＋ T cells and normal CD4＋ T were treated for 48 h with or without ATRA.展开更多
Amelogenin（AMG） is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to inves...Amelogenin（AMG） is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG（rhAMG） on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts（OCCM-30）were treated with different concentrations（0.1, 1, 10, 100, 1 000, 10 000, 100 000 ng · mL^-1） of recombinant human AMG（rhAMG）and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein（BSP）, osteocalcin（OCN）, collagen type I（COL I）, osteopontin（OPN）, runt-related transcription factor 2（Runx2）, cementum attachment protein（CAP）, and alkaline phosphatase（ALP） genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer.Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomalassociated membrane protein-1（LAMP-1）, which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100 000 ng · mL^-1 increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG（100 000 ng · mL^-1） upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100 000 ng · mL^-1 remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10 000 and 100 000 ng · mL^-1 rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue.展开更多
Carboxyl terminus of Hsp70-interacting protein（CHIP or STUB1） is an E3 ligase and regulates the stability of several proteins which are involved in different cellular functions. Our previous studies demonstrated tha...Carboxyl terminus of Hsp70-interacting protein（CHIP or STUB1） is an E3 ligase and regulates the stability of several proteins which are involved in different cellular functions. Our previous studies demonstrated that Chip deficient mice display bone loss phenotype due to increased osteoclast formation through enhancing TRAF6 activity in osteoclasts. In this study we provide novel evidence about the function of CHIP. We found that osteoblast differentiation and bone formation were also decreased in Chip KO mice. In bone marrow stromal（BMS） cells derived from Chip^-/- mice, expression of a panel of osteoblast marker genes was significantly decreased. ALP activity and mineralized bone matrix formation were also reduced in Chip-deficient BMS cells. We also found that in addition to the regulation of TRAF6, CHIP also inhibits TNFα-induced NF-κB signaling through promoting TRAF2 and TRAF5 degradation. Specific deletion of Chip in BMS cells downregulated expression of osteoblast marker genes which could be reversed by the addition of NF-κB inhibitor. These results demonstrate that the osteopenic phenotype observed in Chip^-/- mice was due to the combination of increased osteoclast formation and decreased osteoblast differentiation. Taken together, our findings indicate a significant role of CHIP in bone remodeling.展开更多
Chondrocytes and osteoblasts differentiate from a common mesenchymal precursor, the osteochondroprogenitor（OCP）, and help build the vertebrate skeleton. The signaling pathways that control lineage commitment for OCP...Chondrocytes and osteoblasts differentiate from a common mesenchymal precursor, the osteochondroprogenitor（OCP）, and help build the vertebrate skeleton. The signaling pathways that control lineage commitment for OCPs are incompletely understood. We asked whether the ubiquitously expressed protein-tyrosine phosphatase SHP2（encoded by Ptpn11） affects skeletal lineage commitment by conditionally deleting Ptpn11 in mouse limb and head mesenchyme using ＂Cre-lox P＂-mediated gene excision.SHP2-deficient mice have increased cartilage mass and deficient ossification, suggesting that SHP2-deficient OCPs become chondrocytes and not osteoblasts. Consistent with these observations, the expression of the master chondrogenic transcription factor SOX9 and its target genes Acan, Col2a1, and Col10a1 were increased in SHP2-deficient chondrocytes, as revealed by gene expression arrays, q RT-PCR, in situ hybridization, and immunostaining. Mechanistic studies demonstrate that SHP2 regulates OCP fate determination via the phosphorylation and SUMOylation of SOX9, mediated at least in part via the PKA signaling pathway. Our data indicate that SHP2 is critical for skeletal cell lineage differentiation and could thus be a pharmacologic target for bone and cartilage regeneration.展开更多
The dorsal lingual epithelium,which is composed of taste buds and keratinocytes differentiated from K14^(+)basal cells,discriminates taste compounds and maintains the epithelial barrier.N6-methyladenosine(m^(6)A)is th...The dorsal lingual epithelium,which is composed of taste buds and keratinocytes differentiated from K14^(+)basal cells,discriminates taste compounds and maintains the epithelial barrier.N6-methyladenosine(m^(6)A)is the most abundant mRNA modification in eukaryotic cells.How METTL3-mediated m^(6)A modification regulates K14^(+)basal cell fate during dorsal lingual epithelium formation and regeneration remains unclear.Here we show knockout of Mettl3 in K14^(+)cells reduced the taste buds and enhanced keratinocytes.Deletion of Mettl3 led to increased basal cell proliferation and decreased cell division in taste buds.Conditional Mettl3 knock-in mice showed little impact on taste buds or keratinization,but displayed increased proliferation of cells around taste buds in a protective manner during post-irradiation recovery.Mechanically,we revealed that the most frequent m^(6)A modifications were enriched in Hippo and Wnt signaling,and specific peaks were observed near the stop codons of Lats1 and FZD7.Our study elucidates that METTL3 is essential for taste bud formation and could promote the quantity recovery of taste bud after radiation.展开更多
Methylation of adenosine base on the nitrogen-6 position （N6-methyladenosine, m^6A） is the most common and abundant modification on mRNA transcripts. This post-transcriptional modification was first described in the...Methylation of adenosine base on the nitrogen-6 position （N6-methyladenosine, m^6A） is the most common and abundant modification on mRNA transcripts. This post-transcriptional modification was first described in the 1970s in hepatoma cells （Desrosiers et al., 1974）.展开更多
Methyl-CpG binding protein 2 （MeCP2） has a crucial role in transcriptional regulation and neural development （Ausi6 et al., 2014）. Loss of function mutations of MECP2 in human lead to Rett syndrome （RTT）, a seve...Methyl-CpG binding protein 2 （MeCP2） has a crucial role in transcriptional regulation and neural development （Ausi6 et al., 2014）. Loss of function mutations of MECP2 in human lead to Rett syndrome （RTT）, a severe neurodevelopmental disorders （Amir et al., 1999）, whereas individuals with the chromosomal duplications containing the MECP2 locus showed severe autism-like symptoms （Ramocki et al., 2009）.展开更多
Autophagy is an evolutionarily conserved lysosome-mediated catabolic process（Klionsky,2007）.Autophagy is believed to be essential for cell survival,especially when cells were exposed to stresses,such as nutrient sta...Autophagy is an evolutionarily conserved lysosome-mediated catabolic process（Klionsky,2007）.Autophagy is believed to be essential for cell survival,especially when cells were exposed to stresses,such as nutrient starvation.展开更多
Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in me...Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.展开更多
This report,"Mandible exosomal ssc-mir-133b regulates tooth development in miniature swine via endogenous apoptosis" by Li et al. is an important step forward in describing the factors that control tooth dev...This report,"Mandible exosomal ssc-mir-133b regulates tooth development in miniature swine via endogenous apoptosis" by Li et al. is an important step forward in describing the factors that control tooth development in a large animal model. That many of the regulatory miRNA pathways have been elucidated in murine species have always begged the question as to how relevant they展开更多
Objective:This study was conducted to examine the effect of focal adhesion kinase-associated non-kinase(FRNK)overexpression or inhibition on hepatic stellate cell(HSC)-mediated liver fibrosis progression and to explor...Objective:This study was conducted to examine the effect of focal adhesion kinase-associated non-kinase(FRNK)overexpression or inhibition on hepatic stellate cell(HSC)-mediated liver fibrosis progression and to explore the role of FRNK in improving liver fibrosis.Methods:i)Using primary mouse HSCs and the human HSC cell line LX-2.展开更多
Establishment of a hematopoietic stem cell（HSC）pool depends on the appropriate formation,maturation and mobilization of HSCs in vertebrates.In mice,the aorta-gonad-mesonephros（AGM）is a prominent site for the forma...Establishment of a hematopoietic stem cell（HSC）pool depends on the appropriate formation,maturation and mobilization of HSCs in vertebrates.In mice,the aorta-gonad-mesonephros（AGM）is a prominent site for the formation of definitive HSCs from endothelial cells,although the placenta and yolk sac also give rise to HSCs（Mikkola and Orkin,2006;Chen et al.,2009）.After formation,AGM-derived HSCs migrate to the fetal liver（FL）,and ultimately to the bone marrow （BM）, two definitive hematopoietic organs （Cumano and Godin, 2007）.展开更多
基金supported by Yunnan Province Agricultural Joint Key Project(Grant No.202401BD070001-016)the National Natural Science Foundation of China(Grant No.32202530)+3 种基金Talent Introduction and Training Project of Yunnan Academy of Agricultural Sciences(Grant No.2024RCYP-09)Fundamental Research Project(Grant No.202401CF070046)Xingdian Talent support program(XDYC-QNRC-2023-0457)Yunnan Technology Innovation Center of Flower Technique.
文摘The proper flowering time of rose(Rosa hybrida)is vital for the market value of this horticultural crop,but the mechanism regulating this trait is largely unclear.Here,we found that the transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE4(RhSPL4)positively regulates flowering time in rose.Transient silencing or overexpression transgenic rose plants of RhSPL4 exhibited delayed or early flowering,respectively.Analysis of transcriptome data from transgenic lines overexpressing RhSPL4 compared to the wild type indicated that differentially expressed genes were significantly enriched in the circadian rhythm pathway.Among the proteins encoded by these genes,RhSPL4 binds to the promoter of PSEUDO-RESPONSE REGULATOR 5-LIKE(RhPRR5L),as revealed in yeast one-hybrid,dual-Luciferase/Renilla luciferase reporter,chromatin immunoprecipitation-quantitative PCR and electrophoretic mobility shift assay.Furthermore,RhSPL4 specifically binds to the478 to441 bp region of the RhPRR5L promoter and activates its transcription.The silencing of RhPRR5L delayed flowering time in rose,resembling the phenotype of RhSPL4-silenced plants.Together,these results indicate that the RhSPL4-RhPRR5L module positively regulates flowering time in rose,laying the foundation for the genetic improvement of flowering time in this important horticultural crop.
基金supported by National Natural Science Foundation of China(Grant Nos.32372732,32072586,32372683,32460750,31500247)Natural Science Foundation of Sichuan Province(Grant No.2022NSFSC1689)+3 种基金Sichuan Innovation Team of National Modern Agricultural Industry Technology System(Grant No.SCCXTD-2024-05)Central Leading Local Science and Technology Development Project(Grant No.2023ZYD0090)the Joint research on mustard breeding in Sichuan Province(Grant No.2023YZ002)Guizhou Provincial Key Technology R&D Program[(2021)No.207].
文摘Carotenoid isomerase(CRTISO)is an important enzyme in carotenoid biosynthesis,and it catalyzes the conversion of lycopene precursors to lycopene in several plant species.However,the role of CRTISO in other biochemical processes during plant growth and development remains unclear.Here,we showed that Chinese kale boacrtiso mutants have distinctive characteristics,including a yellowgreen hue and glossy appearance,and this contrasts with the dark green and glaucous traits observed in wild-type(WT)plants.Analysis of pigments in mutants revealed that the reduction in the content of carotenoids and chlorophylls contributed to the yellow-green coloration observed in mutants.An examination of cuticular waxes in Chinese kale indicated that there was a decrease in both the total wax content and the content of individual waxes in boacrtiso mutants(bearing a mutation of BoaCRTISO),which may be caused by the decrease of abscisic acid(ABA)content.The expression of carotenoid,chlorophyll,ABA,and wax biosynthesis genes was downregulated in boacrtiso mutants.This finding confirms that BoaCRTISO regulates the biosynthesis of pigments,ABA,and cuticular waxes in Chinese kale.Our results provide new insights into the interplay between plant pigment and cuticular wax metabolic pathways in Brassica vegetables.
基金National Natural Science Foundation of China(32072531)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS-ASTIP-2016-RIP)+2 种基金the National Key Research and Development Program of China(2021YFE0190700)the ScienceTechnology&Innovation Funding Authority(STDF)of Egypt(43093)for funding this work。
文摘Red fruit peel is one of pear's most valuable economic traits and is mainly determined by anthocyanins.Many pear cultivars with a red peel originated from bud sports;however,little is known about the genetic mechanisms underlying this trait.We have previously identified a mutant Pp BBX24 containing a 14-nucleotide deletion in the coding region(Ppbbx24-del)as the only known variant associated with the red coloration of the mutant‘Red Zaosu'pear(Pyrus pyrifolia White Pear Group).Herein,we analyzed the role of the mutant gene in red coloration and its mechanism of action.The results showed that light promoted red peel coloration in the‘Red Zaosu'pear,and Ppbbx24-del positively affected light-induced anthocyanin biosynthesis,while normal Pp BBX24 had the opposite effects.Transient and stable transformation experiments confirmed that Ppbbx24-del could promote anthocyanin accumulation in pear fruit peels,calli,and tobacco flowers.Due to the loss of nuclear localization sequence(NLS)and viral protein(VP)domains,Ppbbx24-del co-localized in the nucleus and cytoplasm,whereas PpBBX24 localized only in the nucleus.Real-time PCR and transcriptome analyses indicated that PpM YB10 and PpH Y5 are highly expressed in the‘Red Zaosu'pear.In yeast one-hybrid and dual-luciferase assays,Ppbbx24-del and PpHY5 independently promoted the expression of PpC HS,PpC HI,and PpM YB10 by binding to their promoters;however,PpBBX24 did not affect the expression of these genes.Additionally,we found that Ppbbx24-del and PpHY5 had additive effects on the expression of PpC HS,PpC HI,and PpMYB10,as they promote the expression of anthocyanin synthesis genes separately.The co-expression of PpB BX24 and PpHY5 inhibited the activation of downstream genes by PpHY5,which was attributed to the interaction between the two loci.In conclusion,our results clarify the molecular mechanism by which mutant Ppbbx24-del and PpBBX24 exert opposite effects in regulating anthocyanin accumulation in pear.These findings lay an important theoretical foundation for using Ppbbx24-del to create red pear cultivars.
基金supported by the National Natural Science Foundation of China(Grant No.32171851)Natural Science Foundation of Zhejiang(Grant No.LZ23C160003)the Fundamental Research Funds for the Zhejiang A&F University(Grant No.2016FR033)。
文摘Coloration in rose(Rosa hybrida) petals is primarily determined by anthocyanin accumulation in vacuoles, and vacuolar acidification plays a central role in controlling the accumulation of this pigment. Nevertheless, the regulatory interplay between anthocyanin accumulation and tissue acidification processes remains somewhat unclear. The present study characterized an activator Rh MYB114 and a repressor Rh MYB16,which functioned synergistically in anthocyanin accumulation and tissue acidification in rose. Transforming tobacco and roses by overexpression, the introduction of Rh MYB114 resulted in an increase in anthocyanin levels and a noticeable decrease in p H in the petal cells of both rose and tobacco, whereas Rh MYB16 introduction led to inverse effects. To further clarify the underlying the regulatory mechanisms, the yeast two-hybrid(Y2H), bimolecular fluorescence complementation(Bi FC) and dual-luciferase(LUC) were employed. The results showed that Rh MYB16 competed with Rh MYB114, bound to Rhb HLH3 or Rhb HLH33, and inhibited its ability to induce the expression of genes related to anthocyanin biosynthesis and acidification. Our findings revealed a feedback mechanism for the regulation of anthocyanin synthesis and tissue acidification involving Rh MYB114, which stimulated the transcriptional expression of Rh MYB16, whose encoded protein Rh MYB16, in turn, negatively regulated the transcriptional expression of Rh MYB114. Therefore, this study underscores the pivotal roles of the Rh MYB114-Rh MYB16 loop in regulating anthocyanin synthesis and tissue acidification, offering insights into metabolic manipulation to enhance the aesthetic appeal of roses.
基金supported by grants from the Natural Science Foundation of Guangxi Province(Grant No:2022GXNSFAA035639 and 2023GXNSFBA026092)the National Natural Science Foundation of China(Grant No:81860445 and 82260554)the Innovation Project of Guangxi Graduate Education(Grant No:YCBZ2024118)。
文摘Objective Glioma is a highly heterogeneous and malignant intracranial tumor that presents challenges for clinical treatment.ELMO domain containing 2(ELMOD2)is a GTPase-activating protein that regulates a range of cellular biological processes.However,its specific role and prognostic value in tumorigenesis are still unknown.This study aimed to assess the prognostic relevance and signaling function of ELMOD2 in gliomas.Methods The Chinese Glioma Genome Atlas(CGGA)and The Cancer Genome Atlas(TCGA)databases were utilized to conduct a comprehensive analysis of the expression profile of ELMOD2 in gliomas,elucidating its associations with clinicopathological parameters and patient prognosis.Single-cell analysis was performed to characterize ELMOD2 expression across distinct glioma cell subpopulations.Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,and Gene Set Variation Analysis(GSVA)were employed to evaluate the potential biological functions of ELMOD2 in gliomagenesis.Specific small interfering RNAs(siRNAs)were used to knock down ELMOD2 in the glioma cell lines U251 and A172 to assess their cellular behaviors and examine the levels of multiple key signaling molecules associated with the occurrence of gliomas.Results ELMOD2 was overexpressed in gliomas,and this upregulation was correlated with tumor grade,isocitrate dehydrogenase mutation,and 1p/19q codeletion status.Notably,ELMOD2 expression was elevated in classical and mesenchymal subtypes,and single-cell resolution analysis revealed predominant enrichment within malignant cells.Functionally,ELMOD2 regulated cell cycle progression,and its overexpression was related to independent adverse outcomes.In vitro experiments revealed that ELMOD2 was located in the cytoplasm and nucleoplasm.Furthermore,ELMOD2 knockdown reduced proliferation,migration,and invasion and increased apoptosis in U251 and A172 cell lines.Finally,ELMOD2 knockdown significantly decreased p-Erk1/2.Conclusions ELMOD2 expression in glioma is positively correlated with tumorigenesis and is a crucial independent prognostic marker.Thus,ELMOD2 is a promising biomarker and therapeutic target for glioma treatment.
基金supported by the National Natural Science Foundation (NNSF) Key Research Program in Aging (91749204)National Natural Science Foundation of China (81871099, 31370958, 81701364, 81771491, 81501052)+1 种基金Shanghai Municipal Science and Technology Commission Key Program (15411950600, 18431902300)Municipal Human Resources Development Program for Outstanding Leaders in Medical Disciplines in Shanghai (2017BR011)
文摘RANKL signaling is essential for osteoclastogenesis. Its role in osteoblastic differentiation and bone formation is unknown. Here we demonstrate that RANK is expressed at an early stage of bone marrow mesenchymal stem cells(BMSCs) during osteogenic differentiation in both mice and human and decreased rapidly. RANKL signaling inhibits osteogenesis by promoting β-catenin degradation and inhibiting its synthesis. In contrast, RANKL signaling has no significant effects on adipogenesis of BMSCs.Interestingly, conditional knockout of rank in BMSCs with Prx1-Cre mice leads to a higher bone mass and increased trabecular bone formation independent of osteoclasts. In addition, rank: Prx1-Cre mice show resistance to ovariectomy-(OVX) induced bone loss. Thus, our results reveal that RANKL signaling regulates both osteoclasts and osteoblasts by inhibition of osteogenic differentiation of BMSCs and promotion of osteoclastogenesis.
文摘Age-related osteoporosis is associated with the reduced capacity of bone marrow mesenchymal stem cells （BMSCs） to differentiate into osteoblasts instead of adipocytes. However, the molecular mechanisms that decide the fate of BMSCs remain unclear. In our study, microRNA-23a, and microRNA-23b （miR-23a/b） were found to be markedly downregulated in BMSCs of aged mice and humans. The overexpression of miR-23a/b in BMSCs promoted osteogenic differentiation, whereas the inhibition of miR-23a/b increased adipogenic differentiation. Transmembrane protein 64 （Tmem64）, which has expression levels inversely related to those of miR-23a/b in aged and young mice, was identified as a major target of miR-23a/b during BMSC differentiation. In conclusion, our study suggests that miR-23a/b has a critical role in the regulation of mesenchymal lineage differentiation through the suppression of Tmem64.
文摘In the recent two decades, it has been well elucidated that receptor activator of nuclear factor-κB ligand （RANKL; also known as TNFSF11） binding to its receptor RANK （also known as TNFRSF11A） drives osteoclast development as the crucial signaling pathway.;However, accumulating evidence also implies that
基金supported by the National Natural Science Foundation of China(No.30671761)
文摘The essential effect of vitamin A on immune function occurs through various mechanisms including direct effect on ThloTh2 balance modulation. However, it is unclear whether or not vitamin A can regulate Thl-Th2 balance under a strong Thl-polarizing condition. Therefore, the purpose of our study was to examine the effect of vitamin A metabolite allotrans retinoic acid （ATRA） on ThloTh2 differentiation in CD4~ T cells under GATA-3 deficiency, which can induce Thl-polarizing condition. In the present study, GATA-3 deficiency T cells were induced by siRNA and checked by real-time quantitative PCR and western blot. GATA-3 deficiency CD4＋ T cells and normal CD4＋ T were treated for 48 h with or without ATRA.
基金supported by TüBITAK SBAG108S265(Turkey)BMBF TUR08/09(Germany)
文摘Amelogenin（AMG） is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG（rhAMG） on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts（OCCM-30）were treated with different concentrations（0.1, 1, 10, 100, 1 000, 10 000, 100 000 ng · mL^-1） of recombinant human AMG（rhAMG）and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein（BSP）, osteocalcin（OCN）, collagen type I（COL I）, osteopontin（OPN）, runt-related transcription factor 2（Runx2）, cementum attachment protein（CAP）, and alkaline phosphatase（ALP） genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer.Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomalassociated membrane protein-1（LAMP-1）, which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100 000 ng · mL^-1 increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG（100 000 ng · mL^-1） upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100 000 ng · mL^-1 remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10 000 and 100 000 ng · mL^-1 rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue.
基金supported by National Institutes of Health Grants, R01 AR054465, R01 AR070222, and R01 AR070222supported by the grants of Natural Science Foundation of China (NSFC) to TW (grants No. 81301531 and 81572104)+1 种基金supported by the grant from Shenzhen Science and Technology Innovation Committee, China (grant No. JCYJ20160331114205502)the grant from Shenzhen Development and Reform Committee, China for Shenzhen Engineering Laboratory of Orthopedic Regenerative Technologies
文摘Carboxyl terminus of Hsp70-interacting protein（CHIP or STUB1） is an E3 ligase and regulates the stability of several proteins which are involved in different cellular functions. Our previous studies demonstrated that Chip deficient mice display bone loss phenotype due to increased osteoclast formation through enhancing TRAF6 activity in osteoclasts. In this study we provide novel evidence about the function of CHIP. We found that osteoblast differentiation and bone formation were also decreased in Chip KO mice. In bone marrow stromal（BMS） cells derived from Chip^-/- mice, expression of a panel of osteoblast marker genes was significantly decreased. ALP activity and mineralized bone matrix formation were also reduced in Chip-deficient BMS cells. We also found that in addition to the regulation of TRAF6, CHIP also inhibits TNFα-induced NF-κB signaling through promoting TRAF2 and TRAF5 degradation. Specific deletion of Chip in BMS cells downregulated expression of osteoblast marker genes which could be reversed by the addition of NF-κB inhibitor. These results demonstrate that the osteopenic phenotype observed in Chip^-/- mice was due to the combination of increased osteoclast formation and decreased osteoblast differentiation. Taken together, our findings indicate a significant role of CHIP in bone remodeling.
基金supported by NIH R21AR57156NIH R37 CA49152+4 种基金the Rhode Island Hospital Orthopaedic Foundationgrant from the Pediatric Orthopaedic Society of North AmericaArthritis National Research Foundationrecipient of Ryan Fellowshippilot award recipient from NIGMS1P20 GM119943
文摘Chondrocytes and osteoblasts differentiate from a common mesenchymal precursor, the osteochondroprogenitor（OCP）, and help build the vertebrate skeleton. The signaling pathways that control lineage commitment for OCPs are incompletely understood. We asked whether the ubiquitously expressed protein-tyrosine phosphatase SHP2（encoded by Ptpn11） affects skeletal lineage commitment by conditionally deleting Ptpn11 in mouse limb and head mesenchyme using ＂Cre-lox P＂-mediated gene excision.SHP2-deficient mice have increased cartilage mass and deficient ossification, suggesting that SHP2-deficient OCPs become chondrocytes and not osteoblasts. Consistent with these observations, the expression of the master chondrogenic transcription factor SOX9 and its target genes Acan, Col2a1, and Col10a1 were increased in SHP2-deficient chondrocytes, as revealed by gene expression arrays, q RT-PCR, in situ hybridization, and immunostaining. Mechanistic studies demonstrate that SHP2 regulates OCP fate determination via the phosphorylation and SUMOylation of SOX9, mediated at least in part via the PKA signaling pathway. Our data indicate that SHP2 is critical for skeletal cell lineage differentiation and could thus be a pharmacologic target for bone and cartilage regeneration.
基金supported by the National Natural Science Foundation of China(81970913 and 82125006)。
文摘The dorsal lingual epithelium,which is composed of taste buds and keratinocytes differentiated from K14^(+)basal cells,discriminates taste compounds and maintains the epithelial barrier.N6-methyladenosine(m^(6)A)is the most abundant mRNA modification in eukaryotic cells.How METTL3-mediated m^(6)A modification regulates K14^(+)basal cell fate during dorsal lingual epithelium formation and regeneration remains unclear.Here we show knockout of Mettl3 in K14^(+)cells reduced the taste buds and enhanced keratinocytes.Deletion of Mettl3 led to increased basal cell proliferation and decreased cell division in taste buds.Conditional Mettl3 knock-in mice showed little impact on taste buds or keratinization,but displayed increased proliferation of cells around taste buds in a protective manner during post-irradiation recovery.Mechanically,we revealed that the most frequent m^(6)A modifications were enriched in Hippo and Wnt signaling,and specific peaks were observed near the stop codons of Lats1 and FZD7.Our study elucidates that METTL3 is essential for taste bud formation and could promote the quantity recovery of taste bud after radiation.
文摘Methylation of adenosine base on the nitrogen-6 position （N6-methyladenosine, m^6A） is the most common and abundant modification on mRNA transcripts. This post-transcriptional modification was first described in the 1970s in hepatoma cells （Desrosiers et al., 1974）.
基金supported by the Strategic Priority Research Program of Chinese Academy of Sciences(Grant No.XDB02050400)the National Natural Science Foundation of China(Grant No. #91432111) to Z.Qiu
文摘Methyl-CpG binding protein 2 （MeCP2） has a crucial role in transcriptional regulation and neural development （Ausi6 et al., 2014）. Loss of function mutations of MECP2 in human lead to Rett syndrome （RTT）, a severe neurodevelopmental disorders （Amir et al., 1999）, whereas individuals with the chromosomal duplications containing the MECP2 locus showed severe autism-like symptoms （Ramocki et al., 2009）.
基金supported by the National Basic Research Program of China (973 Program)(No.2016YFA0100400)the National Natural Science Foundation of China(No.81773009)
文摘Autophagy is an evolutionarily conserved lysosome-mediated catabolic process（Klionsky,2007）.Autophagy is believed to be essential for cell survival,especially when cells were exposed to stresses,such as nutrient starvation.
基金supported by grants from the National Natural Science Foundation of China(82071150,82170934,81870743,8190104 and 82171001)。
文摘Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.
文摘This report,"Mandible exosomal ssc-mir-133b regulates tooth development in miniature swine via endogenous apoptosis" by Li et al. is an important step forward in describing the factors that control tooth development in a large animal model. That many of the regulatory miRNA pathways have been elucidated in murine species have always begged the question as to how relevant they
文摘Objective:This study was conducted to examine the effect of focal adhesion kinase-associated non-kinase(FRNK)overexpression or inhibition on hepatic stellate cell(HSC)-mediated liver fibrosis progression and to explore the role of FRNK in improving liver fibrosis.Methods:i)Using primary mouse HSCs and the human HSC cell line LX-2.
基金supported by the National Natural Science Foundation of China (No. 81200340)Team Program of Guangdong Natural Science Foundation (No. 2014A030312002)+2 种基金Talent Recruitment fundingExcellent Young Teacher funding of Guangdong Higher Education Institutes (No. Yq2013025)Peal River S&T Nova Program of Guangzhou (No. 2013J2200032)
文摘Establishment of a hematopoietic stem cell（HSC）pool depends on the appropriate formation,maturation and mobilization of HSCs in vertebrates.In mice,the aorta-gonad-mesonephros（AGM）is a prominent site for the formation of definitive HSCs from endothelial cells,although the placenta and yolk sac also give rise to HSCs（Mikkola and Orkin,2006;Chen et al.,2009）.After formation,AGM-derived HSCs migrate to the fetal liver（FL）,and ultimately to the bone marrow （BM）, two definitive hematopoietic organs （Cumano and Godin, 2007）.
