褪黑素对绒山羊皮肤FGF5等绒毛生长相关基因的影响

目的:旨在分析埋植褪黑素后对绒山羊皮肤绒毛生长相关基因表达的影响,进而探讨褪黑素促绒毛生长的机理。方法:以内蒙古白绒山羊为研究对象,选取体况相近的4月龄母羔羊10只,按试验要求随机分为2组,处理组按照2mg/kg BW(体重)的剂量每两个月耳后皮下埋植褪黑素,对照组不埋植,试验时间为1年。实验开始后每月采集绒山羊体侧皮肤和毛样,利用荧光定量PCR技术检测9个绒毛生长相关基因的表达变化。结果:(1)埋植褪黑素后各个基因的平均表达水平都有所上调,其中显著上调FGF5(P<0.0001),而对β-catenin、MSX-2、Wnt10b、BMP2、BMPRIB、NTRK3、Delta1、hairless等无显著影响(P>0.05);(2)埋植褪黑素明显提高了FGF5与hairless的相关性,并且使FGF5与β-catenin、NTRK3的相关性由负相关变成正相关,与Wnt10b的相关性降低,FGF5与其余4个基因的相关性变化不大。结论:结果提示,褪黑素上调绒山羊皮肤FGF的表达,影响FGF5与hairless、β-catenin、NTRK3之间的相关性;山羊中FGF5基因对被毛表型的作用与其他动物不同;褪黑素对复杂组织和单个细胞的作用不同;MAPK信号通路是受褪黑素影响最大的一个信号途径。推测褪黑素可能通过调节绒山羊皮肤中RORα来发挥作用,影响FGF5与hairless、β-catenin、NTRK3和Wnt10b等的相关性,同时抑制MSX2和BMP2的功能,进而促进绒毛的生长。     

Single-Cell Analysis Reveals a Hair Follicle Dermal Niche Molecular Differentiation Trajectory that Begins Prior to Morphogenesis

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Molecular Cloning and Expression Analysis of Prostaglandin E Receptor 2 Gene in Cashmere Goat (Capra hircus) Skin during Hair Follicle Development

As a member of the four subtypes of receptors for prostaglandin E2 (PGE2), prostaglandin E receptor 2 (PTGER2) is in the family of G-protein coupled receptors and has been characterized to be involved in the development and growth of hair follicles. In this study, we cloned and characterized the full-length coding sequence (CDS) of PTGER2 gene from cashmere goat skin. The entire open reading frame (ORF) of PTGER2 gene was 1047 bp and encoded 348 amino acid residues. The deduced protein contained one G-protein coupled receptors family 1 signature, seven transmembrane domains, and other potential sites. Tissue expression analysis showed that PTGER2 gene was expressed strongly in the skin. The general expression tendency of PTGER2 gene at different hair follicle developmental stages in the skin was gradually decreased from anagen to catagen to telogen. After comparing with the expression of BMP4 gene and related reports, we further presume that it seems to have a relationship between the hair follicle cycle and the expression level of PTGER2 gene in cashmere goat skin.     

Azospirillum sp. Promotes Root Hair Development in Tomato Plants through a Mechanism that Involves Ethylene

Tomato seeds were inoculated with the plant growth–promoting rhizobacteria Azospirillum brasilense FT326, and changes in parameters associated with plant growth were evaluated 15 days after inoculation. Azospirilla were localized on roots and within xylematic tissue. An increase in shoot and root fresh weight, main root hair length, and root surface indicated that inoculation with A. brasilense FT 326 resulted in plant growth improvement. The levels of indole-3-acetic acid (IAA) and ethylene, two of the phytohormones related to plant growth, were higher in inoculated plants. Exogenously supplied ethylene mimicked the effect of inoculation, and the addition of an inhibitor of its synthesis or of its physiological activity completely blocked A. brasilense growth promotion. Based on our results, we propose that the process of growth promotion triggered by A. brasilense inoculation involves a signaling pathway that has ethylene as a central, positive regulator.     

用不同模型估计绒山羊早期生长性状遗传参数的比较

利用内蒙古伊盟阿尔巴斯白绒山羊种羊场从1993年至2000年间白绒山羊的早期生长性状(包括出生重、断乳重、日增重和周岁重)的场内测定数据,对4种不同动物模型估计遗传参数的差异进行了比较分析。不同模型中对母体遗传效应和母体环境效应作了不同的考虑：模型I,不考虑母体遗传效应和母体环境效应;模型Ⅱ,仅考虑母体遗传效应;模型Ⅲ,仅考虑母体环境效应;模型Ⅳ,同时考虑母体遗传效应和母体环境效应。利用MTD-FREML程序采用非求导约束最大似然法(DFREML)估计各模型中的方差组分,用似然比检验对不同模型的差异进行检验。结果表明：对于出生重,母体遗传效应和母体环境效应都有极显著的影响,应采用模型Ⅳ进行分析;对于断乳重和日增重,母体遗传效应的影响不显著,而母体环境效应的影响极显著,应采用模型Ⅲ进行分析;对于周岁重,母体环境效应的影响不显著,而母体遗传效应的影响显著,应采用模型Ⅱ进行分析。     

Differential Expression of microRNAs and their Regulatory Networks in Skin Tissue of Liaoning Cashmere Goat during Hair Follicle Cycles

MicroRNAs (miRNAs) are endogenous small noncoding RNA molecules that negatively regulate gene expression. Herein, we investigated a selective number of miRNAs for their expression in skin tissue of Liaoning Cashmere goat during hair follicle cycles, and their intracellular regulatory networks were constructed based on bioinformatics analysis. The relative expression of six miRNAs (mir-103-3p, -15b-5p, 17-5p, -200b, -25-3p, and -30c-5p) at anagen phase is significantly higher than that at catagen and/or telogen phases. In comparison to anagen, the relative expression of seven miRNAs (mir-148a-3p, -199a-3p, -199a-5p, -24-3p, -30a-5p, -30e-5p, and -29a-3p) was revealed to be significantly up-regulated at catagen and/or telogen stages. The network analyses of miRNAs indicated those miRNAs investigated might be directly or indirectly involved in several signaling pathways through their target genes. These results provided a foundation for further insight into the roles of these miRNAs in skin tissue of Liaoning Cashmere goat during hair follicle cycles.     

河西绒山羊GOLA-DRB1基因第2外显子变异特征分析

采用PCR-SSCP、克隆测序等方法,分析600只河西绒山羊GOLA-DRB1基因第2外显子遗传特征、SNPs变异类型,同时分析该基因与山羊流产的关联性.结果表明,河西绒山羊DRB1基因第2外显子上存在26个等位基因,序列比对后发现26个等位基因中存在71个核苷酸变异住点,占分析位点总数的30％.其中转换位点25个,占核苷酸多态位点的35.21％;颠换位点35个,占核苷酸多态位点的49.3％;转换和颠换共存位点9个,占12.68％.山羊流产关联性分析结果表明,在河西绒山羊病例组中DRB1*18、DRB1*23、DRBl*26等位基因频率高于正常对照组(P＜0.05),x2值分别为6.31,4.859,6.396,RR值为2.55, 2.72,DRB1*8等位基因频率显著高于正常对照组(P＜0.01),x2值为17.618,RR值为3.59;病例组中DRB1*14等住基因频率低于正常对照组(P＜0.05),x2值为4.812,RR值为0.65,DRB1*1和DRB1*11显著低于正常对照组(P＜0.0l),其中B1的x2值和RR值分别为14.11和0.61,初步推断DRB1*8可能是河西绒山羊流产发病单体型中的遗传易感基因,DRB1*1和DRB1*11可能为其遗传保护基因.     

辽宁新品系绒山羊血液蛋白多态性及其与经济性状的关系&

利用聚丙烯酰胺凝胶电泳法对120只辽宁新品系绒山羊的6种血液蛋白进行多态性检测,并分析了蛋白标记座位与体重、绒产量和绒细度等经济性状的相关性。采用SAS统计软件处理,统计各经济性状的最小二乘均值（LSM）,并进行方差分析和多重比较。结果表明：EsBB型可作为体重性状的优势基因型。TfBB和Amy1-2型位点可作为绒产量的优势基因型,可望作为遗传标记用于标记辅助选择。     

The role of Sox9 in maintaining the characteristics and pluripotency of Arbas Cashmere goat hair follicle stem cells

In our previous work, we isolated Arbas Cashmere goat hair follicle stem cells (gHFSCs) and explored the pluripotency. In this study, we investigated the expression and putative role of Sox9 in the gHFSCs. Immunofluorescence staining showed that Sox9 is predominantly expressed in the bulge region of the Arbas Cashmere goat hair follicle, and also positively expressed in both nucleus and cytoplasm of the gHFSCs. When the cells were transfected using Sox9-shRNA, cell growth slowed down and the expression of related genes decreased significantly, cell cycle was abnormal, while the expression of terminal differentiation marker loricrin was markedly increased; cells lost the typical morphology of HFSCs; the mRNA and protein expression of gHFSCs markers and stem cell pluripotency associated factors were all significantly decreased; the expression of Wnt signaling pathway genes LEF1, TCF1,c-Myc were significantly changed. These results suggested that Sox9 plays important role in gHFSCs characteristics and pluripotency maintenance.     

Dexpanthenol Promotes Cell Growth by Preventing Cell Senescence and Apoptosis in Cultured Human Hair Follicle Cells

Dexpanthenol (D-panthenol) is a precursor of vitamin B5 (pantothenic acid) and is widely used for dietary supplements and topical applications. D-panthenol has long been used in hair care products for the purpose of anti-hair loss, its effects and the underlying mechanisms, however, were barely reported. In this study, the effects of D-panthenol on human hair follicle cells, including dermal papilla cells (hDPCs) and outer root sheath cells (hORSCs), were investigated. D-panthenol enhanced the cell viability, increasing the cellular proliferation marker Ki67 in cultured hDPCs. The markers for apoptosis (Caspase3/9) and cell senescence (p21/p16), reported to be expressed in aged or resting phase follicles, were significantly reduced by D-panthenol. Anagen-inducing factors (ALP; β-catenin; versican), which trigger or elongate the anagen phase, were stimulated by D-panthenol. On the other hand, D-panthenol reduced TGF-β1 expressions in both mRNA and protein levels. The expression of VEGF, which is important for peripheral blood vessel activation; was up-regulated by D-panthenol treatment. In cultured hORSCs, cell proliferation and viability were enhanced, while the mRNA expression of cell senescence markers (p21/p16) was significantly down-regulated. The expressions of both VEGF and its receptor (VEGFR) were up-regulated by D-panthenol. In conclusion, our data suggest that the hair growth stimulating activity of D-panthenol was exerted by increasing the cell viability, suppressing the apoptotic markers, and elongating the anagen phase in hair follicles.     

纳米硒对岢岚绒山羊妊娠母羊及胎儿抗氧化性、硒蛋白表达及生长发育的影响

为研究纳米硒对岢岚绒山羊妊娠母羊及胎儿抗氧化能力、硒蛋白表达和生长发育的影响。选择体重相近的岢岚妊娠绒山羊80只,随机分为2组,分别喂以基础日粮和添加0.5mg/kg DM纳米硒的饲粮,实验期110d。结果表明:日粮中纳米硒的添加极显著(P<0.01)地提高了母羊及胎儿血清中GSH-Px、SOD的活性;极显著(P<0.01)提高了母羊及胎儿肝、胎盘组织中GSH-Px、SOD的活性;极显著(P<0.01)降低了母羊及胎儿血清与胎盘组织中MDA的浓度。在日粮中添加纳米硒后,母羊及胎儿肝、肾组织中cGPx1 mRNA的表达显著提高(P<0.05);胎盘组织中的cGPx、SeP、Trx1 mRNA的表达极显著提高(P<0.01)。妊娠母羊的基础日粮中加入纳米硒,极显著(P<0.01)提高了妊娠母羊及胎儿血清、肝、胎盘组织中IGF-1的含量和IGF-1mRNA表达量,显著增加了胎盘、胎儿及胎儿肝、肾组织的重量(P<0.05)。日粮中添加0.5mg/kg DM纳米硒,增强了妊娠母羊及胎儿的抗氧化能力,提高了IGF-1、cGPx1、SeP和Trx1 mRNA的表达量,进一步促进了胎儿的生长发育。     

山羊FGF5基因组织表达及其变异与羊绒性状的关联性分析

为了研究山羊FGF5基因的遗传变异和组织表达等分子特征及其变异对山羊羊绒性状的影响,本试验应用PCR-SSCP与测序相结合的方法检测FGF5基因第1外显子在陇东绒山羊、柴达木绒山羊和中卫山羊中的多态性,并分析该基因变异对羊绒性状的影响;采用RT-qPCR技术研究FGF5基因在陇东绒山羊和辽宁绒山羊组织中的表达.结果 表明,在山羊FGF5基因的第1外显子区检测到一个同义突变位点c.272C＞T,表现为MM、MN和NN3种基因型,其中NN型仅在陇东绒山羊中有极少量分布.3个山羊品种的优势基因型均为MM,优势等位基因均为M.c.272C＞T位点在3种山羊品种中均处于Hardy-Weinberg平衡状态,且均为低度多态.c.272C＞T位点与陇东绒山羊的羊绒性状关联性分析表明,该位点对羊绒纤维直径的影响显著.携带等位基因N的个体的羊绒纤维直径显著低于不含等位基因N的个体的(P＜0.05);基因型MN个体的羊绒纤维直径显著低于基因型MM个体的(P＜0.05).FGF5基因在陇东绒山羊和辽宁绒山羊的心、肝、脾、肺、肾、骨骼肌和皮肤中均有表达;在陇东绒山羊的心、脾、肺、肾和皮肤中的表达量显著或极显著高于辽宁绒山羊的.本研究结果提示,若以改善羊绒细度为选育目标,山羊FGF5基因可作为羊绒性状改良的候选基因.     

Genome-Wide Analysis Reveals Novel Regulators of Growth in Drosophila melanogaster

Organismal size depends on the interplay between genetic and environmental factors. Genome-wide association (GWA) analyses in humans have implied many genes in the control of height but suffer from the inability to control the environment. Genetic analyses in Drosophila have identified conserved signaling pathways controlling size; however, how these pathways control phenotypic diversity is unclear. We performed GWA of size traits using the Drosophila Genetic Reference Panel of inbred, sequenced lines. We find that the top associated variants differ between traits and sexes; do not map to canonical growth pathway genes, but can be linked to these by epistasis analysis; and are enriched for genes and putative enhancers. Performing GWA on well-studied developmental traits under controlled conditions expands our understanding of developmental processes underlying phenotypic diversity.     

绒山羊Scp3基因的克隆及睾丸中第一轮减数分裂的发生

联会复合体蛋白3（synaptonemal complex protein 3, Scp3）是雄性生殖细胞减数分裂中联会复合体形成必需的组成部分,在哺乳动物生殖细胞减数分裂中具有重要功能。通过RT-PCR扩增和分子克隆的方法首次克隆到了绒山羊Scp3基因的编码区片段。测序结果表明,绒山羊与牛的Scp3基因编码序列同源率达到98％。根据测得的DNA序列,设计引物,用RT-PCR方法分析测定了青春前期绒山羊不同个体中睾丸组织的Scp3的转录表达。结果显示,雄性绒山羊青春前期睾丸发育存在个体差异,已分析的样品第一轮减数分裂发生的时间普遍为73天之后。这一结果为绒山羊的睾丸发育和精子发生过程的相关研究打下了基础。     

成年绒山羊皮肤干细胞克隆定向诱导分化为成骨细胞

成年绒山羊皮肤经0.02%中性蛋白酶4℃过夜消化,分离表皮,37℃消化30min,经100μg/mlIV型胶原处理的培养皿黏附10min筛选干细胞,培养到第二代时采用有限稀释法进行单细胞克隆纯化,角蛋白15、角蛋白19免疫荧光染色部分细胞强阳性,添加成骨细胞诱导液定向诱导分化,经碱性磷酸酶、茜素红染色鉴定为阳性,表明分离的细胞是皮肤干细胞且有能力诱导分化为成骨细胞。     

Molecular Analysis of a Deletion Hotspot in the NRXN1 Region Reveals the Involvement of Short Inverted Repeats in Deletion CNVs

NRXN1 microdeletions occur at a relatively high frequency and confer increased risk for neurodevelopmental and neurobehavioral abnormalities. The mechanism that makes NRXN1 a deletion hotspot is unknown. Here, we identified deletions of the NRXN1 region in affected cohorts, confirming a strong association with the autism spectrum and other neurodevelopmental disorders. Interestingly, deletions in both affected and control individuals were clustered in the 5′ portion of NRXN1 and its immediate upstream region. To explore the mechanism of deletion, we mapped and analyzed the breakpoints of 32 deletions. At the deletion breakpoints, frequent microhomology (68.8%, 2–19 bp) suggested predominant mechanisms of DNA replication error and/or microhomology-mediated end-joining. Long terminal repeat (LTR) elements, unique non-B-DNA structures, and MEME-defined sequence motifs were significantly enriched, but Alu and LINE sequences were not. Importantly, small-size inverted repeats (minus self chains, minus sequence motifs, and partial complementary sequences) were significantly overrepresented in the vicinity of NRXN1 region deletion breakpoints, suggesting that, although they are not interrupted by the deletion process, such inverted repeats can predispose a region to genomic instability by mediating single-strand DNA looping via the annealing of partially reverse complementary strands and the promoting of DNA replication fork stalling and DNA replication error. Our observations highlight the potential importance of inverted repeats of variable sizes in generating a rearrangement hotspot in which individual breakpoints are not recurrent. Mechanisms that involve short inverted repeats in initiating deletion may also apply to other deletion hotspots in the human genome.