哺乳动物性别决定机理及性别鉴定方法研究进展

哺乳动物的性别决定是由SRY基因为调控中心、多基因参与的级联调控过程。本文综述了性别决定基因及其功能、性别决定的分子模型及性别鉴定的方法等方面的研究进展。     

家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

Effect of live weight and dietary supplement of raw potato starch on the levels of skatole, androstenone, testosterone and oestrone sulphate in entire male pigs

This study evaluated the effect of supplement of raw potato starch (RPS) on the levels of skatole, androstenone, testosterone and oestrone sulphate in plasma from entire male pigs. The study also evaluated relationships between plasma levels of skatole and testicular steroids at three different live weights (LW) of approximately 90, 100 and 115 kg. A total of 111 entire male pigs of a crossbred (Yorkshire dams×Swedish Landrace sires) were used. Animals were raised either in mixed pens, with females and males, or single-sex pens. Each pen contained seven or nine pigs. The most fast-growing three pigs from the pens with nine pigs were slaughtered when they reached 90 kg LW, and the remaining pigs were slaughtered at 115 kg LW. All pigs were fed the same commercial diet until the average pen weight reached 100 kg. Then, 33 out of 80 remaining pigs received RPS, 0.6 kg per pig and day, for 2 weeks prior to slaughter. Blood samples were taken from the pigs at three occasions: first, the day prior to first slaughter occasion, low-weight group; second, the day prior to change in diet, middle-weight group; and third, the day prior to second slaughter occasion, high-weight group. Plasma was analysed for the levels of skatole, androstenone, testosterone and oestrone sulphate. Fat samples were taken at slaughter and analysed for the levels of skatole and androstenone. The levels of skatole and testicular steroids in plasma were significantly higher in entire male pigs from the high-weight group fed no RPS compared to those from low- and middle-weight groups. The levels of the investigated compounds did not differ between low- and middle-weight groups (P>0.1). The diet with RPS induced a decline in skatole levels in plasma and fat (P<0.001), but not plasma levels of testicular steroids and fat levels of androstenone (P>0.05). Skatole levels were positively correlated to testosterone and oestrone sulphate levels in the middle- and high-weight pigs fed no RPS as well as to testosterone in the low-weight group. In the high-weight group fed RPS, skatole levels were not correlated to any of the analysed compounds. Approximately 26% of the entire male pigs (11 out of 43) from the high-weight group fed no RPS produced skatole levels in fat above 0.20 μg/g, whereas the pigs from the low- and high-weight group fed RPS did not produce skatole levels above 0.20 μg/g in fat. Androstenone levels in fat were high in all groups. In total 47% (52 out of 111) pigs expressed androstenone levels above the rejection levels in fat of 1.0 μg/g and 88% (98 out of 111) had androstenone levels above 0.5 μg/g. It was concluded that a lower slaughter weight and the supplement of raw potato starch to the diet could be used to reduce skatole levels in entire male pigs. Androstenone levels in fat, however, could not be reduced by either a lower weight at slaughter or dietary manipulation.     

Quick detection of sex determining region protein gene in mouse fetuses

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.     

罹患口腔炎乌苏里蝮的相关生理学指标分析

口腔炎是人工养殖蛇类的常见病之一,为了深入了解和尽早做好预防工作,对罹患口腔炎的成年雄蛇乌苏里蝮(Gloydius ussuriensis)进行解剖观察,并检测了相关器官的系数、血液学指标和激素等。结果表明:整体肥满度、躯体肥满度、肾系数等数值罹患口腔炎蛇略高于健康蛇;而肝系数、精巢系数和脂肪体系数等罹患口腔炎的蛇低于健康雄蛇,且前两者呈显著性差异,后者呈极显著性差异。性激素检测结果显示罹患口腔炎的蛇血液睾酮和雌二醇含量均低于健康蛇,且均具极显著性差异。血细胞指标中,红细胞数量、红细胞分布均差、白细胞总数、淋巴细胞数量和淋巴细胞比率等均为罹患口腔炎的蛇高于健康雄蛇,且具显著性差异,但嗜中性粒细胞和淋巴细胞的比值(NLR)、中性粒细胞数量和比率、中间细胞数量及其比率均为患口腔炎的雄蛇低于健康蛇,且具显著性差异。结论:口腔炎不仅影响了主要代谢器官的器官系数的变化,而且血液多项指标和性激素水平也有明显差异。性激素含量的变化将对乌苏里蝮的正常生殖繁育产生不可忽视的影响。另外,NLR和红细胞分布宽度2个血常规指标可作为早期诊断炎症发生的首选参数。     

Effect of mixed rearing of barrows and gilts on backfat thickness and serum metabolite profiles of the Kagoshima-Kurobuta (Berkshire) pig

We investigated the effect of mixed rearing of barrows and gilts on the backfat thickness and the serum metabolite profiles of Kagoshima-Kurobuta (Berkshire) pigs. A total of 149 pigs with an average body weight of 35 kg were divided into the following groups: 100%, 90%, 70%, 50%, 30%, 10%, and 0% groups consisting of 10 barrows (1 pen), 9 barrows + 1 gilt (3 pens), 7 barrows + 3 gilts (2 pens), 5 barrows + 5 gilts (3 pens), 3 barrows + 7 gilts (2 pens), 1 barrow + 9 gilts (3 pens), and 9 gilts (1 pen), respectively. All pigs were raised to a shipping weight of 120 kg. Mixed rearing significantly reduced (p < 0.001) backfat thickness, and the optimum mixing ratio of barrows and gilts was 7:3 (the 70% group). Four types of circulating sex steroids were found in both the barrows and gilts in the 50% group but were not detected in barrows from the 100% group. These results indicated that mixed rearing of barrows and gilts was effective for reducing the backfat thickness of barrows, and induced sex steroid hormones may influence the backfat thickness of barrows in mixed-reared groups.     

人工养殖西伯利亚狍肠道微生物组成的雌雄差异

肠道微生物是一个复杂的生态系统,与宿主的健康、疾病以及其他因素有着密切的关系。目前,关于肠道菌群结构和功能的研究较多,但是对西伯利亚狍肠道菌群的研究还未见报道。西伯利亚狍(Capreolus pygargus)属于反刍动物,有独特的消化特性和微生物群来帮助它们适应高纤维的食物。本研究利用高通量测序技术在Illumina MiSeq平台上对14只狍肠道微生物进行16S rRNA基因分析。结果表明:个体样本中门类组成的差异,雌性个体JY3和雄性个体JY9除Firmicutes,Bacteroidetes,Proteobacteria这3类菌群外,其他门类菌群所占比例都很低。在科水平上,雌性个体中Ruminococcaceae在JY7和JY13肠道内含量相对较少,Bacteroidaceae在JY7,JY10和JY13中含量较高。雄性个体中Ruminococcaceae在JY11和JY14肠道内含量较少,Prevotellaceae在JY4中含量较少。在属水平上,雌性个体特有的菌属为Roseburia,Alloprevotella,Lachnospiraceae_incertae_sedis和Coprococcus;雄性个体特有的菌属为Fusobacterium,Succiniclasticum,Streptococcus和Clostridium sensu stricto。Alpha多样性分析表明,Ace指数范围2128.93—772.98,Chao1指数范围2128.89—785.27,Shannon指数范围5.44—3.78,Simpson指数范围0.07—0.01。在不同物种分类水平上,雌雄狍肠道内的优势菌群有些不同。PCoA(principal co-ordinates analysis)分析显示,狍样本几乎混在一起,没有明显差异。     

Effects of different lighting on reproductive system in depressive female rats

AIM: To investigate the effects of different lighting on the reproductive system in depressive female rats. METHODS: Healthy adult female rats were randomly chosen as control group, and the depressive adult female rats in SPF grade were randomly divided into 5 groups(7 rats each):depressive model group, sulfur lamp group, heat radiation lamp group, fluorescent lamp group and LED lamp group. After 45 d of continuous illumination, the estrous cycle was observed by the vaginal exfoliated cells, and the organ indexes of ovary and uterus were calculated. The concentrations of estiadrol(E₂), prolactin(PRL), progesterone(PROG) and testosterone(T) in the serum were detected by ELISA, and the histopathological lesion of ovary was observed under microscope with HE staining. RESULTS: The estrous cycle exhibited serious disorder, the ovaries exhibited serious congestion, and the organ indexes of ovary and uterus and the concentrations of E₂, PRL, PROG and T decreased significantly in the rats in depressive model group compared with control group(P<0.05). The estrous cycle and histopathological damage of ovary were obviously improved, and the concentrations of E₂, PRL, PROG and T were significantly increased after the sulfur lamp lighting in the depressive female rats compared with depressive model group. No obvious change and improvement of the reproductive functions in the heat radiation lamp group, fluorescent lamp group and LED lamp group was observed. CONCLUSION: The reproductive functions of depressive female rats are improved by sulfur lamp lighting.     

赤眼蜂寄生行为研究（Ⅳ）—子代数量分配和子代性分配

本文在对赤眼蜂性控行为研究的基础上〔3〕,进一步探讨了不同种赤眼蜂在同一种寄主不同分布以及同一蜂种(松毛虫赤眼蜂 Trichogramma dendrolimi)在不同种寄主上的子代数量分配和子代性分配问题。研究结果表明,赤眼蜂具有根据寄主卵体积大小分配及调整其子代数量和子代性别的能力,在不同体积的寄主上具有明显不同的子代数量安排策略和子代性安排策略。在同一种寄主卵上,不同种的赤眼蜂对子代性分配的安排存在着较明显的种间差异.在米蛾卵上,松毛虫赤眼蜂的第一雄卵主要在其第一产卵序产出,稻螟赤眼蜂的第一雄卵主要在其第二产卵序产出,拟澳洲赤眼蜂产第一雄卵的位置比较分散,主要分布在前三卵序。松毛虫赤眼蜂大约每产6粒雌卵而产1粒雄卵;稻螟赤眼蜂和拟澳洲赤眼蜂大约每产5.33和3.37粒雌卵而产1粒雄卵(见表2).寄主卵的体积不同,雌蜂在不同寄主上所分配的子代数量也明显不同。松毛虫赤眼蜂在每粒米蛾卵上只安排1粒蜂卵;在每粒马尾松毛虫卵上平均安排18.64粒蜂卵;在每粒“人造卵”上平均安排33.15粒,在柞蚕卵上平均安排77.43粒蜂卵(见表3)。随着寄主卵体积的增大,雌蜂产第一雄卵的时间后移,在松毛虫卵上,松毛虫赤眼蜂雌蜂在产下第一粒雄卵后,大约以每产16粒雌卵而产1粒雄卵;在柞蚕卵上,大约每隔15粒雌卵产1粒雄卵;在“人造卵”上,产雄卵的间隔大约为9粒雌卵。     

氮素形态不同配比对黄瓜生长和性别分化的影响（初报）

采用基质培养研究氮素形态不同配比对黄瓜生长及性别分化的影响,结果表明用纯NO3--N营养液处理,黄瓜植株营养生长旺盛,当NH4 -N占25％和50％时,黄瓜的生殖生长明显促进,表现为比纯NO3--N处理第一雌花低0.4、0.6节,雌花率增加了23.9％、31.0％,单株产量增加了35.3％、28.7％。叶片和果实中矿物元素钾增加了0.02％～1.41％,铁增加了11～124,ug/g DW,锌增加了37.6～47.0μg/g DW,镁、锰、铜差异不大。黄瓜在不同生育期对氮素形态的敏感性有差异,幼苗期比生长旺盛期不敏感。