Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.     

草地贪夜蛾种群性诱测报方法研究

草地贪夜蛾已经成为中国农业生产的重大害虫,准确地预测预报是指导防治的基础。本研究采用国产草地贪夜蛾性诱剂诱陷多测试了3种不同类型诱捕器对草地贪夜蛾的田间诱捕效果。结果表明,诱陷多有效期可达60 d以上,3种诱捕器中以桶型诱捕器诱捕量最高。实验室研究显示,不同日龄草地贪夜蛾雄虫精巢长轴长度有显著性差异,精巢长轴长度随日龄增加不断减小。据此规律,制定了按照精巢长轴大小判断雄蛾日龄的指标,并反演重构了田间雌、雄虫种群羽化的动态曲线。本研究表明,可以利用性诱到的雄虫的精巢发育状况推算田间种群的年龄结构,进而达到通过预测雌虫产卵与幼虫孵化动态指导防治的目的。     

Effect of a single injection of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) on testicular blood flow measured by color doppler ultrasonography in male Shiba goats

Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= −0.862; PI, r= −0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= −0.610) and PI (r= −0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= −0.718; hCG, r= −0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.     

单环刺螠精巢年周期发育及精子发生

采用组织学方法观察了单环刺螠(Urechis unicinctus)精巢的形态结构及其年周期发育,并利用超微技术观察了精子的发生过程。结果表明,单环刺螠的精巢呈条带状,位于虫体尾部,两端通过肌束分别与呼吸肠管壁外侧和体壁内侧相连。精巢由生精细胞团与结缔组织组成,结缔组织位于精巢中央,精原细胞团分布于结缔组织外周,精母细胞团散布于结缔组织内,各级精母细胞脱离精巢落入体腔液中,历经精母细胞、精细胞,最后形成精子脱离细胞团进入肾管。根据精巢的组织学特征,将单环刺螠的精巢年周期发育划分为增殖期、生长期、成熟期、排放期和休止期5个时期。光镜和电镜下观察了精巢和体腔液中精子发生各时期的结构变化,包括细胞核形态的变化、顶体发生以及尾部的形成。本研究揭示了单环刺螠的精巢发育特点、精子发生历经的场所和分化特征。     

不同时期小鼠睾丸合成睾酮能力的差异

不同发育阶段,睾丸合成睾酮的能力差异很大。为了阐明引起睾酮合成差异的分子机制,实验以不同年龄的SPF昆白小鼠,采用RT-PCR与Western blot分析了不同年龄阶段类固醇合成酶的变化。结果显示:(1)在不同年龄阶段,P450scc(胆固醇支链裂解酶)mRNA、3β-HSD(3β-羟胆固醇脱氢酶)mRNA、StAR(类固醇合成快速调节蛋白)mRNA和蛋白的水平无明显变化;(2)P450c17(C17-20裂解酶)mRNA的表达水平在30、60和120d均处于较高的水平,但是在270d则处于较低的水平;(3)30~270d,细胞外信号调节激酶(ERK)活性无明显变化。表明,P450c17 mRNA表达差异可能导致不同时期睾酮合成能力差异的重要原因,但ERK活性可能与衰老引起睾酮合成能力下降无直接关系。     

用免疫荧光技术检测细胞培养物中兔出血症病毒

用本实验室制备的兔出血症病毒（rabbit hemorrhagic disease virus,RHDV）高免血清,按常规方法提纯出抗体（IgG）,用异硫氰酸荧光素（FITC）标记IgG。在细胞培养时,培养瓶中放入玻片,当细胞长成单层时,按常规方法接种病毒液,培养24、48、96、120h时取出玻片,用荧光抗体染色,在荧光显微镜下观察不同代次的细胞毒。经观察：兔肾上皮细胞（RK）毒培养到36—48h,羊睾丸细胞（ST）毒培养到72—96h时可观察到特异性荧光,随着培养时间的延长,荧光亮度增强,胞浆内充满特异性荧光。用肝组织强毒病料触片,呈特异性荧光,对照细胞培养48h无荧光出现,证实了两株细胞培养物中有大量的兔出血症病毒存在,从而成功的分离培养出了兔出血症病毒细胞毒。     

獭兔睾丸组织形态观察

獭兔是一种典型的经济动物,它既是毛皮动物又可食用,普遍应用于人类生产生活。目前已有研究报道了多种哺乳动物生精周期研究,但很少有关于獭兔的报道。本实验对2只完全性成熟的健康獭兔进行睾丸结构,生精周期相对频率,支持细胞效率研究。獭兔睾丸经石蜡包埋,HE染色以便进行组织形态学观察。不同的生殖细胞组合分析显示30%的曲精小管显示两个或两个以上的生精上皮周期阶段。獭兔生精周期阶段频率(%)分别为:阶段1:(15.5±2);阶段2:(14.0±1);阶段3:(11.5±1);阶段4:(8.5±1);阶段5:(10.5±1);阶段6:(18.5±2);阶段7:(13.0±1);阶段8:(8.5±1)。支持细胞效率(圆形精子:支持细胞=7:1)。本研究结果表明獭兔的生精效率较其他哺乳动物高。     

Study on Expression of Bcl-2 and PCNA Genes in Testicular Tissue of Chickens,Quails and Chicken-quail Hybrids

In order to investigate the molecular mechanism of stunted growth testis of chicken (♂) and quail (♀) hybrids. 84 testicular tissue samples of New Roman cocks, male Korean quails and chicken-quail hybrids at different developmental stages were collected,the mRNA expression of Bcl-2 and PCNA genes in testicular tissue of cocks, quails and chicken-quail hybrids at different growth stages were detected using Real-time PCR. The results showed that the Bcl-2 and PCNA genes mRNA expression patterns in testes of chickens and quails at different growth stages were similar.The Bcl-2 gene was fluctuated in a whole, and decreased to the lowest value, then recovered rapidly and remained at a high level. The PCNA gene had a very significant peak value.Compared with the chicken and quail,the expression of Bcl-2 and PCNA genes mRNA in hybrids had no obvious change, which indicated that the pattern of the Bcl-2 gene regulation apoptosis in testis was different from chicken and quail at the hybrid testicular development process, and there was no obvious cell appreciation process.It was suggested that the molecular influencing factors of testicular dysplasia of chicken and quail hybrids were related to the abnormal expression of Bcl-2 and PCNA genes mRNA in testes.The results provided an important reference for the further study of the mechanism of testicular dysplasia between chicken and quail interspecific hybrids.     

关中奶山羊睾丸不同发育阶段代谢物的差异分析

【目的】 试验旨在从代谢组学角度探究关中奶山羊睾丸发育代谢机制。【方法】 选择1月龄断奶雄性关中奶山羊和24月龄体成熟的雄性关中奶山羊各6只,采集睾丸组织,使用液相色谱-质谱（LC-MS）技术对睾丸组织进行化学检测,通过多维和单维分析相结合的方式对两组不同代谢物进行化学检测、对比与鉴别,通过富集差异代谢物分析筛选关中奶山羊发育代谢中的潜在关键通路。【结果】 在1和24月龄关中奶山羊睾丸组织中共筛选出334个差异代谢物,其中显著上调137个,显著下调197个。对前36个差异代谢物进行比较鉴定发现,分别为脂质和类脂质分子、有机酸及其衍生物、未分类化合物和苯丙烷和聚酮化合物四大类。经相关性分析发现,在1和24月龄关中奶山羊睾丸发育过程中,脂质和类脂质分子与甘油磷脂呈最大正相关,羧基及其衍生物与甘油磷脂呈最大负相关。对不同的代谢物进行富集分析,筛选出7条潜在的重要代谢通路,分别为肿瘤中的胆碱代谢、精氨酸和脯氨酸代谢、肿瘤中的中心碳代谢、铁死亡、谷胱甘肽代谢、氨酰-tRNA生物组成以及牛磺酸和亚牛磺酸代谢。【结论】 本试验从代谢组学角度揭示了1和24月龄关中奶山羊睾丸的差异代谢物及代谢机制,为今后哺乳动物睾丸的发育研究奠定了基础。