动物性别决定的分子机理及性别鉴定与控制新技术

哺乳动物中位于Ｙ染色体短臂临界区域的ＳＲＹ基因启动雄性性状的发育。该基因在生殖腺脊中的表达可激发下游基因ＭＬＳ的转录,引起缪氏体抑制物和睾酮分泌,促使睾丸组织器官的发育。针对该基因制备特异探针或产该区域片段得到引物对就可用ＦＩＳＨ或ＰＣＲ准确,快速鉴定出植入母体前的胚胎性别,以及对精子筛选分离的结果作出准确评价。与ＰＣＲ相比,ＦＩＳＨ技术更具检测优势。     

肝素或钙离子载体诱导牛、羊冻精体外获能的研究

本文采用肝素或钙离子载体(I-A)诱导牛、羊冻精体外获能,根据对去透明带仓鼠卵的穿透情况评定获能效果,并用台盼蓝·姬姆萨(TG)染色观察了精子的死活及顶体状态.结果发现,牛、羊冻精经肝素或I-A处理后与卵子一起培养,2 h穿透仓鼠卵,4 h开始形成雄原核,6 h形成发育良好的雄原核.牛冻精用50μg/mL肝素及0.1μmol/L I-A处理效果最好,穿透率分别为72.8％和92.3％;羊则以20μg/mL肝素及0.3μmol/L I-A为宜,穿透率分别为73.1％和68.O％.咖啡因与I-A或肝素并用均有协同作用,能提高精子的穿透能力,并能促进卵内原核的发育.肝素和I-A均能在体外诱导牛、羊冻精的顶体反应,有顶体反应活精子百分率与穿透率呈强正相关,羊冻精比较脆弱,获能处理后活力较低.     

Multiple paternity,reproductive skew and correlates of male reproductive success in a wild population of the Trinidadian guppy

Abstract – In most species, males have a higher reproductive potential than females, leading to skewed reproductive success, particularly in mating systems where pre‐ or postcopulatory sexual selection reinforces inequality in male mating success. We investigated multiple paternity, reproductive skew and correlates of male reproductive success in a wild population of the Trinidadian guppy (Poecilia reticulata). We used nine microsatellite loci to assess the frequency of multiple paternity, number of sires per brood and reproductive skew. Across broods, the frequency of multiple paternity was high with 94% of broods having multiple sires (range: 1–5), resulting in a reproductive skew of 0.14. Variation in male reproductive success was high (range: 0–14 offspring per male), suggesting that there is considerable opportunity for sexual selection. Next, we examined correlations between male reproductive success and sexual coloration, sperm velocity and gonopodium length. Relative area of orange, black, iridescent and total coloration, and sperm velocity were not correlated with reproductive success. However, gonopodium length explained 14% of the variation in reproductive success, suggesting that gonopodium length is likely a sexually selected trait. We discuss these findings in the light of other studies that genetically dissect joint‐sex parentage and examine correlates of male reproductive success in wild populations.     

SMALL INTESTINAL EMPTYING TIME IN NORMAL BEAGLE DOGS

Gastric emptying time and small intestinal transit time in dogs are frequently discussed. However, it is often of interest to the radiologist to know what normal small intestinal emptying times should be. A total of 15 upper gastrointestinal studies was performed on five internal parasite-free, normal, standard Beagle dogs with three studies on each dog, 6 days apart. The ages and weights of the dogs ranged from 2–8 years and from 12.4–13.7 kg, respectively. Following 24-hour fasting, a dose of 10 ml/kg bw of 60% wt/vol barium sulfate suspension was administered through a stomach tube. Then, sequential radiographs were made at 30-minute intervals until the entire contrast medium column was in the colon and cecum. The mean, standard deviation, and range of gastric emptying time, small intestinal transit time, and small intestinal emptying time were 76 ± 16.7 (30–120), 73 ± 16.4 (30–120), and 214 ± 25.1 (180–300) minutes, respectively. This study offers the possibility that small intestinal emptying time may be used to further evaluate patients with suspected small intestinal partial obstruction, pseudo-obstruction, ischemia, or lymphangiectasia.     

冻存液添加蜂王浆主蛋白对精子品质的影响

目的 精液冷冻保存作为人工授精不可或缺的一个技术环节,对优质畜禽种群的繁衍和保存有着至关重要的意义。目前,因精液冻存时冻存液成分、冷冻方法和外部氧化应激等因素影响,导致冻存精液品质不一。蜂王浆（RJ）已被证明能提高动物精液品质,蜂王浆主蛋白（MRJPs）作为RJ主要生物活性成分物质,具有多种生物活性和抗氧化能力。方法 为提高冻存精子品质,本研究开展了在公牛精液冻存液中添加不同浓度（0 g/25 mL、0.01 g/25 mL、0.02 g/25 mL、0.03 g/25 mL、0.04 g/25 mL）的MRJPs冻干粉,对冻存48 h后解冻精子的总活力、前进性活力、畸形率等相关参数进行了观察评估。结果 添加MRJPs呈浓度依赖性方式显著降低精子总活力、快速前进性活力、缓慢前进性活力和直行性活力（P<0.05）,而且精子畸形率和精子原地移动活力与对照组相比无显著差异（P> 0.05）。结论 精液冻存液中添加MRJPs会抑制冻存精子活力,因此,对MRJPs能以何种方式提高精液品质的研究还需要进一步开展。     

miRNA在哺乳动物配子发生中的作用研究进展

哺乳动物配子发生的基因表达调控包括编码基因的阶段特异性表达调控及非编码基因的转录和转录后水平调控。微小RNA(microRNA, miRNA)作为一类小的非编码RNA, 通过识别靶基因非翻译区的结合位点, 导致mRNA降解或者蛋白质翻译抑制, 从而在转录后水平发挥调控作用。近年来, miRNA在哺乳动物生殖活动中的作用逐渐被揭示, 越来越多的研究表明, miRNA在哺乳动物精子发生、精子成熟、卵母细胞成熟、卵泡发育及早期胚胎发育等过程中都发挥着重要的调节作用, 其可通过调节支持细胞的增殖和凋亡或精原细胞、精母细胞及精细胞的细胞周期进程, 在精子发生的不同阶段发挥间接或直接调控作用, 也可通过调节卵母细胞、卵丘细胞以及颗粒细胞的增殖、凋亡、激素合成和细胞间作用, 对卵母细胞的发育和成熟过程进行调控。作者主要介绍了在哺乳动物配子发生过程中, miRNA的细胞和阶段特异性表达及其对靶基因的调节作用, 以期为深入研究哺乳动物配子发生的调控机制提供参考。     

Evidence of LAT1 expression in canine caput epididymis

L-type amino acid transporter 1 (LAT1), the first isotype of amino acid transport system L, transports aromatic and branched amino acids pivotal for fundamental cellular activities such cellular growth and proliferation. LAT1 expression was high only in the brain in contrast to its limited distribution and low level of expression in normal tissues. We found potent LAT1 expression in canine caput epididymis by quantitative RT-PCR and Western blotting analysis. Immnuno-histochemical examination revealed observable LAT1 in microvillous epithelial cells.     

Effect of lactoferrin on murine embryo development created from lipopolysaccharide-treated sperm

The effect of lactoferrin (LF) on embryo development was investigated by using lipopolysaccharide (LPS)-treated mouse sperm. For the development rate of the 2-cell stage embryo, the embryo derived from LPS- and LF-treated sperm showed similar survival rate to the control embryo. On day 12 after the embryo transfer into the recipient, the frequent abnormality was observed in the embryo derived from LPS-treated sperm, and the abnormality was tended to be inhibited in the embryo derived from LPS- and LF-treated sperm. These results imply that LF treatment on sperm contaminated with bacteria may facilitate the embryo development, which contribute to the improvement of infertility.     

嗜水气单胞菌双组分系统FlrBC的功能探究

为明确双组分系统FlrBC在嗜水气单胞菌极生鞭毛合成、生物被膜形成及致病机制中的作用,通过同源重组法,以嗜水气单胞菌ZYAH72为野生菌株构建双组分系统FlrBC缺失株ΔflrB和ΔflrC,比较不同菌株鞭毛合成及游动性、生物被膜形成、胞外多糖分泌、细胞黏附以及抗全血杀伤能力差异。结果显示:与野生株相比,ΔflrB和ΔflrC仍能形成极生鞭毛,细菌游动能力无显著差异;而结晶紫染色发现ΔflrB和ΔflrC相较于野生株的生物被膜形成能力分别下降了27.2%和22.3%;刚果红检测结果显示,与野生株相比,ΔflrB和ΔflrC的胞外多糖分泌分别下降了18.4%和14.2%;实时荧光定量PCR的结果显示,flrB和flrC的缺失不同程度抑制了鞭毛合成、生物被膜相关通路基因的表达;与草鱼肾细胞共孵育后,ΔflrB和ΔflrC的细胞黏附率与野生株相比分别下降了23.2%和18.2%;全血杀伤实验结果显示,ΔflrB和ΔflrC抗全血杀伤能力均显著减弱。以上结果表明,双组分系统FlrBC不是嗜水气单胞菌极生鞭毛形成所必需,但在细菌鞭毛形成组装和生物被膜形成中发挥调控作用,并且影响嗜水气单胞菌致病性...