Increases in litter size, which are influenced by ovulation, are responsible for between 74% and 96% of the economic value of genetic progress, which influences selection. For the selection and breeding of highly prolific goats, genetic mechanisms underlying variations in litter size should be elucidated. Here, we used single-nucleus RNA sequencing to analyze 44,605 single nuclei from the ovaries of polytocous and monotocous goats during the follicular phase. Utilizing known reference marker genes, we identified 10 ovarian cell types characterized by distinct gene expression profiles, transcription factor networks, and reciprocal interaction signatures. An in-depth analysis of the granulosa cells revealed three subtypes exhibiting distinct gene expression patterns and dynamic regulatory mechanisms. Further investigation of cell-type-specific prolificacy-associated transcriptional changes elucidated that “downregulation of apoptosis”, “increased anabolism”, and “upstream responsiveness to hormonal stimulation” are associated with prolificacy. This study provides a comprehensive understanding of the cell-type-specific mechanisms and regulatory networks in the goat ovary, providing insights into the molecular mechanisms underlying goat prolificacy. These findings establish a vital foundation for furthering understanding of the molecular mechanisms governing folliculogenesis and for improving the litter size in goats via molecular design breeding.

MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate several pathway intermediates and affect the skeletal muscle development in mice, pigs, sheep, and cattle. However, to date, only a small number of miRNAs have been reported in the muscle development of goats. In this report, the longissimus dorsi transcripts of one- and ten-month-old goats were analyzed by sequencing RNAs and miRNAs. The results showed that the ten-month-old Longlin goats had 327 up- and 419 down-regulated differentially expressed genes (DEGs) compared with the one-month-old. In addition, 20 co-up-regulated and 55 co-down-regulated miRNAs involved in the muscle fiber hypertrophy of goats were identified in ten-month-old Longlin and Nubian goats compared with one-month-old. Five miRNA-mRNA pairs (chi-let-7b-3p-MIRLET7A, chi-miR193b-3p-MMP14, chi-miR-355-5p-DGAT2, novel_128-LOC102178119, novel_140-SOD3) involved in the goat skeletal muscle development were identified by miRNA-mRNA negative correlation network analysis. Our results provided new insight into the functional roles of goat muscle-associated miRNAs, allowing a deeper understanding of the transformation of miRNA roles during mammalian muscle development.

Resveratrol (RES) is one of the best-known bioactive polyphenols that has received much attention in recent years because of its importance to anti-obesity. However, the exact mechanism underlying this effect and whether it can improve lipid metabolism by regulating the long-chain noncoding RNA (lncRNA) remains unclear. In this study, 24 healthy crossbred castrated boars were fed a basal diet (control) and a basal diet supplemented with 200 mg, 400 mg or 600 mg RES per Kilogram (kg) of feed for 41 d, respectively. We found that 400 mg/kg and 600 mg/kg RES-supplemented diet did not affect growth rate, but reduced significantly subcutaneous adipose thickness, carcass fat rate, greater dramatically the serum concentration of adiponectin and high-density lipoprotein in pigs. Further, we verified that RES could inhibit the formation and accumulation of lipid droplets by AdipoQ-AdipoR1-AMPKα and AdipoQ-AdipoR2-PPARα signal pathway in vivo and vitro (3T3-L1 preadipocytes). Transcriptome analyses found that 5 differently expressed (DE) lncRNAs and 77 mRNAs in subcutaneous adipose between control group and 400 mg/kg RES group, which mainly involved in "adipocytokine signaling pathway," "Wnt signaling pathway," "PI3K-Akt signaling pathway" and "MAPK signaling pathway." In conclusion, RES can inhibit the formation and accumulation of lipid droplets through AdipoQ signal pathway and lipid metabolism-related lncRNAs. Our results provide a new insight on the molecular mechanism of RES as a nutritional agents to the prevention and treatment for obesity.

This study investigated the effects of resveratrol (RES) supplementation on the growth performance, carcass and meat quality, blood lipid levels and ruminal bacterial microbiota of fattening goats. A total of forty castrated Nubian goats (28.25 ± 0.26 kg body weight) were randomly divided into four groups and provided with diets containing different levels of RES (0, 150, 300 and 600 mg/kg) for 120 d. The results showed that RES increased redness and intramuscular fat content, whilst reducing shear force in the longissimus dorsi muscle of goats (p < 0.05). In addition, the final weight, average daily gain, hot carcass weight, net meat weight, carcass lean percentage and eye muscle area of goats were significantly increased in the 150 mg/kg RES group compared with the other three groups, while those in the 600 mg/kg RES group significantly decreased (p < 0.05). RES significantly decreased serum triacylglycerol and LDL-C contents (p < 0.05), and increased HDL-C content and the HDL-C/TC ratio (p < 0.05). Supplementation with 150 mg/kg RES also increased the proportion of Acetitomaculum and Moryella, genera comprising short-chain fatty acid-producing bacteria. The present study indicated that an appropriate supplemental level of RES could improve the growth performance, neat percentage, meat quality, ruminal microbiota and serum lipid levels of fattening goats.

试验旨在研究饲粮中添加白藜芦醇(resveratrol,RES)对"杜×长×大"猪空肠抗氧化酶活性及抗氧化基因指标和紧密连接蛋白相关基因表达水平的影响.试验选用12头体重约为65 kg的育肥猪分为2组,每组6头.对照组育肥猪饲喂基础饲粮,试验组育肥猪饲喂在基础饲粮中添加400 mg/kg的RES的饲粮,试验期41 d.结果 表明,与对照组相比,饲粮中添加400 mg/kg的RES显著提高了育肥猪空肠过氧化氢酶(CAT)、总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性及总抗氧化能力(T-AOC) (P＜0.05),显著降低丙二醛(MDA)的含量(P＜0.05);显著提高育肥猪空肠抗氧化基因铜/锌超氧化物歧化酶(Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-SOD)、谷胱甘肽过氧化物酶-1(GPx-1)和谷胱甘肽过氧化物酶-4 (GPx-4)活性(P＜0.05);显著提高育肥猪空肠抗氧化酶活相关基因CAT、 T-SOD和GSH-Px的表达水平(P＜0.05),显著降低MDA的表达水平(P＜0.05);显著提高育肥猪空肠紧密连接蛋白咬合蛋白(Occludin)、闭合蛋白-1(Claudin-1)和链接黏附分子A (JAM-A)的表达水平(P＜0.05).研究表明,饲粮添加400mg/kg的RES能够显著提高育肥猪空肠抗氧化能力及紧密连接蛋白的表达,改善动物肠道健康.

[目的]阐明山羊瘤胃微生物特征及品种间的差异,为进一步研究不同山羊品种生长性能与瘤胃微生物群间的关系提供参考依据.[方法]随机挑选都安山羊和努比亚山羊健康母羊各3只,正常饲喂6个月后屠宰,采集经4层纱布过滤的瘤胃液2 mL,采用16S rDNA扩增子高通量测序技术分析都安山羊和努比亚山羊的瘤胃微生物区系特征及品种间的差异.[结果]都安山羊和努比亚山羊瘤胃微生物在门分类水平上的组成相似,均以厚壁菌门、拟杆菌门和变形菌门为优势菌群,在都安山羊样品中的相对丰度分别为58.32％、29.41％和4.26％,在努比亚山羊样品中的相对丰度分别为45.66％、38.69％和6.59％.在属分类水平上,以奎因氏菌属、普氏菌属_1和理研菌科RC9肠道群为优势菌群,在都安山羊样品中的相对丰度分别为36.93％、9.67％和12.07％,在努比亚山羊样品中的相对丰度分别为12.91％、13.92％和11.47％.品种间瘤胃差异菌种分析结果表明:在门分类水平上,都安山羊瘤胃中不可培养菌_k_细菌的相对丰度极显著低于努比亚山羊(P<0.01,下同),黏胶球形菌门的相对丰度显著低于努比亚山羊(P<0.05,下同);在属分类水平上,都安山羊瘤胃中奎因氏菌属的相对丰度极显著高于努比亚山羊,不可培养菌_k_细菌的相对丰度极显著低于努比亚山羊,而Pyramidobacter、疣微菌科_UCG-010、Proable_genus_10、不可培养菌_f_p-251-o5、不可培养菌_f_vadinBE97、不可培养菌_o_拟杆菌、疣微菌科_UCG-002、Flexilinea、脱硫弧菌属、丁酸弧菌属及Mogibacterium的相对丰度显著低于努比亚山羊.PICRUSt2的微生物群落功能预测发现,都安山羊和努比亚山羊的瘤胃微生物在碳水化合物代谢及其他氨基酸代谢和神经系统等功能存在显著差异.[结论]都安山羊和努比亚山羊的瘤胃微生物优势菌群相似,但其微生物群落组成、多样性及功能存在一定差异,尤其是植物性纤维降解相关微生物丰度差异显著,可能对山羊具有较强适应性发挥重要作用.

Resveratrol (RES) has a wide range of biological and pharmacological activities with various health benefits for humans as a food additive. In animal production, RES has been considered a potential functional feed additive for producing high-quality pork. Long noncoding RNAs (lncRNAs) have emerged as essential regulators of fat metabolism, and phytochemicals can regulate fat metabolism through lncRNA. However, it is unclear whether RES can improve back-fat thickness by regulating lncRNA. In this study, we identified a novel lncRNA, which was named a long intergenic non-protein coding RNA, a regulator of fat metabolism (LincRNA-ROFM), from our previous lncRNA sequencing data. LincRNA-ROFM can inhibit adipocyte proliferation and differentiation. In-depth analyses showed that LincRNA-ROFM acts as a molecular sponge for miR-133b, and adiponectin (AdipoQ) is a direct target of miR-133b in porcine preadipocytes. In addition, the expression of LincRNA-ROFM was positively correlated with AdipoQ. RES can promote the expression of LincRNA-ROFM by PPARα and C/EBPα. Altogether, our research showed that LincRNA-ROFM acts as a ceRNA to sequester miR-133b and is upregulated by RES, leading to heightened AdipoQ expression, and thus decreased adipocyte proliferation and differentiation, which reduces back-fat thickness of pigs. Taken together, the RES/LincRNA-ROFM/miR-133b/AdipoQ regulatory network preliminarily explains the mechanism of action of RES in inhibiting fat deposition, which provides new insight into the downstream mechanism of RES inhibition of fat deposits by regulating the lncRNA.

[目的]明确转移抑制素基因(KISS1)多态性与努比亚山羊繁殖性状(产羔数、初生重和断奶重)的关联,为进一步展开山羊分子标记辅助选择育种提供参考依据.[方法]利用DNA混合池结合Sanger测序筛选出努比亚山羊KISS1基因SNP位点,再通过MTA-Seq测序对355只努比亚山羊群体进行SNP位点的基因分型,并采用SAS 9.2的一般线性模型(GLM)对KISS1基因SNP位点与努比亚山羊的产羔表型性状进行关联分析.[结果]在努比亚山羊KISS1基因内含子1发现2个SNP位点(g.1341600A>G和g.1341674C>G).其中,g.1341600A>G位点处于中度多态(0.25<PIC<0.50),且处于Hardy-Weinberg平衡状态(PHWE>0.05);而g.1341674C>G位点处于低度多态(PIC<0.25),已偏离Hardy-Weinberg平衡状态(PHWE<0.05).g.1341600A>G位点GG和AG基因型个体的第一胎、第二胎及三胎联合产羔数均高于AA基因型个体,且在第一胎达显著水平(P<0.05,下同);g.1341674C>G位点CC基因型个体则表现为第一胎、第三胎及三胎联合产羔数均高于GG基因型个体,同样在第一胎达显著水平.g.1341674C>G位点GG基因型个体的第一胎、第三胎及三胎联合羔羊初生重均高于CC基因型个体,且在第三胎达显著水平;g.1341600A>G位点不同基因型与努比亚山羊各胎次羔羊初生重无显著关联(P>0.05,下同).在羔羊断奶重方面,2个SNP位点不同基因型与努比亚山羊各胎次断奶重也无显著关联.g.1341600A>G位点和g.1341674C>G位点可形成连锁不平衡单倍型模块,其中GC、AC、AG单倍型的频率分别为0.645、0.265和0.090;单倍型模块与第一胎产羔数显著关联,AACC组合型产羔数显著低于其他组合型.[结论]努比亚山羊KISS1基因内含子1存在2个SNP位点(g.1341600A>G和g.1341674C>G)与其繁殖性状相关,其中g.1341600A>G位点的A等位基因和g.1341674C>G位点的C等位基因可作为努比亚山羊选育的潜在分子标记,且在今后的育种过程中应淘汰KISS1基因AACC组合型母羊.

研究旨在对努比亚山羊脂肪和肥胖相关蛋白(fat mass and obesityassociated protein,FTO)基因进行克隆和分析,并构建其真核表达载体.取努比亚山羊背最长肌组织作为试验材料,采用RT-PCR扩增出FTO基因的编码区,测序鉴定后对得到的序列用相应的分析软件进行生物信息学分析,然后将FTO基因片段与pMD19-T载体连接后转化大肠杆菌DH5α感受态细胞,构建pMD19-T-FTO载体,测序正确的重组质粒双酶切后,连接pEGFP-N1载体构建pEGFP-N1-FTO真核表达载体,然后转染3T3-L1细胞,培养48 h后,在荧光显微镜下观察细胞表达荧光的情况.结果表明,试验成功克隆了努比亚山羊FTO基因的编码区,序列长度为1518 bp,编码505个氨基酸,分子质量为57142.24 u.努比亚山羊FTO基因编码区序列与NCBI上公布的山羊、牛、绵羊、猪、原鸡、小鼠的相似性分别为98.7％、96.4％、98.3％、87.2％、64.3％、81.7％,该基因系统进化树分析显示,努比亚山羊与山羊遗传距离最近,与原鸡遗传距离最远,在不同物种中具有高度保守性.努比亚山羊FTO蛋白二级和三级结构以α-螺旋和无规则卷曲为主.构建的真核表达载体pEGFP-N1-FTO转染3T3-L1细胞48 h后,在显微镜下观察到绿色荧光的表达,说明FTO基因真核表达载体构建成功.本试验构建努比亚山羊FTO基因真核表达载体,其在3T3-L1细胞中成功表达,为以后研究FTO基因与山羊脂肪代谢的相关性奠定基础.

雌性动物卵泡发育是一个动态的、受众多激素及调控因子直接或间接参与的复杂生物学过程,其中卵巢颗粒细胞的增值和分化能调节卵泡的生长和成熟,颗粒细胞的凋亡,影响卵泡闭锁.环氧合酶-2(cyclooxygenase 2,COX-2)在卵巢中主要在颗粒细胞中表达,可通过对下游代谢产物的调节,以不同的方式抑制颗粒细胞凋亡,参与PI3 K/AKT/NOTCH/Wnt信号通路的调节,从而作为排卵诱导因子诱导排卵.

Heat stress (HS) results in health problems in animals. This study was conducted to investigate the effect and the underlying mechanism of HS on the proliferation and differentiation process of 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were treated at 37 °C or 41.5 °C. HS up-regulated the mRNA and protein expression level of heat shock protein 70 (HSP70). Furthermore, the proliferation of 3T3-L1 preadipocytes were significantly inhibited after HS treatment for 2 days. A large number of accumulated lipid droplets were observed under the microscope after HS treatment for 8 days. Notably, the result of oil red O staining showed that the number of lipid droplets increased significantly and the differentiation ability of the cells was enhanced after HS. Moreover, after 2 and 8 d of differentiation, HS increased the transcription levels of fat synthesis genes including peroxisome proliferators activated receptor γ (PPARγ), fatty acid binding protein 2 (AP2), fatty acid synthase (FAS) and CCAAT enhancer binding protein α (CEBPα) genes, while decreasing the transcription levels of lipid decomposition genes including ATGL and HSL genes. In addition, HS reduced the expression of AMPK and PGC-1α, as well as the dephosphorylation of AMPK. 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR) can eliminate HS induced lipogenesis by activating AMPK. These results indicated that HS inhibited the proliferation of 3T3-L1 preadipocytes and promoted lipid accumulation by inhibiting the AMPK-PGC-1α signaling pathway in 3T3-L1 preadipocytes. This work lays a theoretical foundation for improving the effect of HS on meat quality of livestock and provides a new direction for the prevention of obesity caused by HS.

[目的]明确隆林山羊诱导细胞凋亡DFF45样效应因子c基因(CIDEc)的生物学特性及其表达规律,为揭示CIDEc基因对山羊脂肪代谢的调控机制提供理论依据.[方法]提取隆林山羊背最长肌、心脏、肝脏、脾脏、肾脏、腹脂和皮下脂肪及努比亚山羊腹脂和皮下脂肪的总RNA,PCR扩增隆林山羊CIDEc基因编码区(CDS)序列,使用Ex-PASy、TMHMM Server v.2.0、ProtScale、NPS@SOPMA和SWISS-MODEL等在线软件进行生物信息学分析,并利用实时荧光定量PCR检测CIDEc基因在隆林山羊和努比亚山羊不同组织器官中的表达情况.[结果]隆林山羊CIDEc基因CDS序列全长为741 bp,共编码244个氨基酸残基,其编码蛋白分子量26.09 kD,不稳定系数48.44,脂肪指数100.7,理论等电点(pI)5.28,属于酸性蛋白,不存在跨膜结构,亲水性较强.隆林山羊CIDEc基因CDS序列与NCBI已公布的山羊CIDEc基因(XM_018038446.1)CDS序列相对比仅有1处碱基发生突变,即第560位点G突变为T,属于同义突变.隆林山羊与绵羊的CIDEc基因CDS序列相似性最高(99.0％),与小鼠的相似性较低(79.6％);基于CIDEc基因CDS序列相似性构建的系统发育进化树也显示隆林山羊与绵羊的亲缘关系最近,与小鼠的亲缘关系较远.在隆林山羊CIDEc蛋白的二级结构中:α-螺旋占32.38％,β-转角占12.70％,延伸链占13.11％,无规则卷曲占41.80％.CIDEc基因在隆林山羊7个组织器官中均有表达,且在腹脂和皮下脂肪中高表达,极显著高于在其他器官组织的相对表达量(P<0.01);CIDEc基因在努比亚山羊脂肪组织(皮下脂肪和腹脂)中的相对表达量显著高于在隆林山羊脂肪组织中的相对表达量(P<0.05).[结论]CIDEc基因在隆林山羊各器官组织中均有表达,以在脂肪组织中的表达水平较高,且其在努比亚山羊脂肪组织(皮下脂肪和腹脂)中的表达水平显著高于在隆林山羊中的表达水平.可见,CIDEc蛋白是脂肪代谢的重要调节剂,与动物体内脂质存储密切相关.

Litter size is an important component trait of doe reproduction. By improving it, production efficiency and economic benefits can be significantly provided. Genetic marker-assisted selection (MAS) based on proven molecular indicators could enhance the efficacy of goat selection, as well as litter size trait. Many molecular markers have been identified that they can be used to improve litter size in different goat breeds. However, the presence and value of these markers vary among goat breeds. In the present study, we used the reported loci on other breeds of goat as candidate loci to detect whether these loci appear in this Nubian goat population; then we proceed to genotype and detect surrounding loci (50 bp) by multiplex PCR and sequencing technology. As a result, 69 mutations (59 SNPs and 10 indels) were screened out from 23 candidate genes in Nubian goat population, 12 loci were significantly associated with the litter size of first-parity individuals; 5 loci were significantly associated with the litter size of second-parity individuals; 3 loci were significantly associated with the litter size of third-parity individuals. In addition, five loci were significantly associated with the average litter size. The additive effect value of KITLG: g.18047318 G>A in first parity, KITLG: g.18152042G>A in third parity, KISS-1: g.1341674 C>G in first parity, and GHR: g.32134187G>A in second parity exceed more than 0.40, and the preponderant alleles are G, C, A and G, respectively. Further, linkage disequilibrium analysis of 21 mutation loci shows that 3 haplotype blocks are formed, and the litter size of combination type AACC in KISS-1 gene and AAGG in KITLG gene are significantly lower than that of other combinations genotype in first parity (P<0.05). These findings can provide effective candidate DNA markers for selecting superior individuals in Nubian goat breeding.

为了探讨山羊诱导细胞凋亡DFF45样效应因子a(CIDEa)基因分子特征,了解其组织表达谱和在不同品种山羊之间的表达差异,试验对隆林山羊CIDEa基因进行克隆和生物信息学分析,采用实时荧光定量PCR方法检测CIDEa基因的组织表达情况.结果 表明:克隆获得的隆林山羊CIDEa基因编码区(CDS区)全长为660 bp,编码219个氨基酸,蛋白质分子质量为23 367.21 ku,属于亲水性不稳定膜外蛋白.与普通山羊CIDEa基因CDS区对比,隆林山羊CIDEa基因序列存在一个核苷酸突变位点,由亮氨酸变为脯氨酸.隆林山羊CIDEa基因与绵羊CIDEa基因同源性最高,与猪、猕猴、人的同源性较低,与大鼠、小鼠的遗传距离最远;隆林山羊CIDEa基因编码蛋白中α-螺旋占35.62％,T-转角占8.68％,β-氨基酸残基构成的延伸链占16.44％,无规则卷曲占39.27％;隆林山羊CIDEa基因在腹脂和皮下脂肪中高表达,其表达水平显著高于努比亚山羊,在其他组织中表达水平极低或不表达.说明试验成功克隆出隆林山羊CIDEa基因,该基因与山羊脂肪代谢有着密不可分的关系.

在山羊产业中,窝产仔数是极其重要的经济性状,但潜在遗传分子机制尚不清楚.文章综述了山羊窝产仔数在DNA水平、RNA水平以及蛋白质水平的研究进展,以期为今后对山羊窝产仔数差异的理解和研究提供帮助,并为山羊品种的资源开发、基因挖掘、品种选育、遗传改良和种质特异性评价提供一定的理论依据.

[目的]获取正常形态下广西巴马小型猪心血管系统、运动系统、呼吸系统、中枢神经系统和泌尿系统的电子计算机断层扫描(CT)影像图谱及各组织器官的测量数据,为广西巴马小型猪影像数据库建立及其后续研究提供参考依据.[方法]选取10头健康合格的12月龄广西巴马小型猪,公母各半,全身麻醉后置于CT诊断台上,采用头前尾后俯卧姿势,腹部正中矢状面垂直于扫描床平面并与床面长轴的中线重合,从头顶至足部连续进行扫描,并采用多平面重建(MPR)、最大密度投影(MIP)和容积再现技术(VR)等观察分析广西巴马小型猪各系统的解剖关系.[结果]将广西巴马小型猪原始容积数据进行图像重组,即可获得心血管、运动、呼吸、中枢神经和泌尿系统等五大系统的三维重建图像,同时测量获得各系统主要组织器官的相关数据,且发现公猪和母猪间差异不显著(P>0.05);但在获得的广西巴马小型猪CT影像图谱中肌肉、细小血管及神经等显示并不清晰,无法测量,致使各系统中的组织器官测量数据并不完全.[结论]通过CT技术对正常形态下的广西巴马小型猪进行全身影像观察及各组织器官数据测量,可实现不用屠宰解剖(活体)即能观察其内部结构,使广西巴马小型猪解剖学及疾病模型应用更直观.

We present a new piezoelectric flow energy harvester (PFEH) based on a piezoelectric cantilever (PEC) with a cylindrical extension. The flow-induced vibration of the cylindrical extension causes the PEC to vibrate at the natural frequency of the PFEH. The PFEH provides a low-cost, compact, and scalable power source for small electronics by harvesting energy from ambient flows such as wind and water streams. Prototypes were tested in both laminar and turbulent air flows, demonstrating the feasibility of the design. Turbulence excitation was found to be the dominant driving mechanism of the PFEH with additional vortex shedding excitation contribution in the lock-in region.