不同繁殖季节成年牦牛附睾组织特征比较

为比较不同繁殖季节成年牦牛附睾组织结构特征变化,应用苏木精-曙红常规染色、Masson's和Gomori's特殊组织化学染色方法比较不同繁殖季节牦牛（6头繁殖期成年牦牛和9头繁殖间期成年牦牛）附睾的组织结构特点并用IPP图像分析软件进行定量分析。结果显示,与繁殖间期相比,繁殖期附睾尾间质胶原纤维和网状纤维较附睾头及附睾体明显增多;附睾头和附睾体柱状上皮厚度显著增加（P<0.05）,附睾头纤毛长度增加显著（P<0.05）;附睾头和附睾尾管腔内径及外径显著增高（P<0.05）;但是附睾尾管腔外径繁殖期与繁殖间期相比差异不显著（P>0.05）。本研究结果表明,成年牦牛附睾管外围网状纤维与胶原纤维分布一致,二者在附睾尾较为丰富,可能与其较强的收缩能力及精子运输有关;柱状纤毛上皮高度及纤毛长度、管腔内径与外径的变化与其所处的不同繁殖季节密切相关,附睾管腔的膨大和回缩亦可能是高原地区季节性繁殖的哺乳动物中一种普遍存在的现象。     

热应激小鼠附睾组织HSP70表达的研究

利用化学恒温培养箱对小鼠实施42℃1h/d的热应激处理,建立持续的全身热应激动物模型。在热应激持续到8d、13d、21d和35d时,颈椎脱臼处死对照组和热应激组小鼠,应用改良巴氏染色法检测附睾精子畸形率,免疫荧光组织化学和蛋白质印迹分析的方法检测附睾组织HSP70的表达。结果显示：热应激组小鼠附睾精子畸形率随着热应激持续时间的延长而升高,且显著（P<0.05）高于对照组;对照组小鼠附睾组织HSP70呈极强表达（+++）,而热应激组小鼠附睾组织HSP70的表达随着热应激持续时间的延长而减弱,当热应激持续到35d时呈极弱的表达（±）。结果表明：热应激损伤了附睾的功能,使附睾精子畸形率增加;附睾内HSP70为非热诱导型,HSP70可能在附睾精子的成熟过程发挥特殊的作用。     

Correlation of testicular artery Doppler velocimetry with kinetics and morphologic characteristics of epididymal sperm in dogs

Sperm quality can be affected by a reduction in testicular blood flow, which can be measured by Doppler ultrasonography. The aim of this study was to correlate the Doppler velocimetry of the testicular artery with kinetics of the epididymal spermatozoa in dogs. Twenty-two dogs (44 testicles) were evaluated by Doppler ultrasonography in five regions of the testicular artery before orchiectomy. Spermatozoa were recovered by the epididymal tail compression technique and analysed for kinetics on a computer-assisted semen analysis (CASA system). Morphology (modified Karras) and sperm membrane integrity were analysed by eosin–nigrosine staining. Data were analysed by Pearson's correlation test (p < .01). The mean total motility was 69.0% ± 17.7, progressive motility was 43.7% ± 14.7, average path velocity (VAP) was 127.0 µm/s ± 20.7, curvilinear velocity (VCL) was 221.0 µm/s ± 31.1, and sperm velocity index (SVI) was 389.9 ± 56.1. There were positive correlations between the peak systolic velocity (PSV) in the proximal supratesticular region with the SVI (r = .529), VCL (r = .555) and VAP (r = .473), and a negative correlation with the percentage of slow spermatozoa (r = −.463). The results suggest that the testicular artery blood flow velocity can positively affect the speed of spermatozoa movement. For the first time, we have correlated sperm kinetics with the Doppler evaluation of the testicular artery in dogs.     

RBP4基因在绵羊性成熟前后附睾中表达的研究

试验旨在研究视黄醇结合蛋白4（retinol binding protein 4,RBP4）基因在绵羊性成熟前和性成熟后附睾中的表达水平,探讨其在绵羊生殖机能调控方面的作用。以绵羊性成熟前和性成熟后附睾为材料,采用实时荧光定量PCR方法检测RBP4基因mRNA在绵羊性成熟前、后附睾中表达量的差异,采用免疫组化技术检测RBP4基因在绵羊性成熟前、后附睾头和附睾尾中蛋白的表达水平。结果显示,RBP4基因在绵羊附睾中性成熟前的表达量为1.3368,性成熟后的表达量为0.6450,虽然二者间的表达量差异不显著（P>0.05）,但性成熟前的表达量高于性成熟后的表达量。RBP4蛋白在性成熟前、后附睾头上皮层、平滑肌层、组织间质中均有阳性表达,主要位于上皮细胞质、平滑肌细胞质;在附睾尾上皮层、组织间质中有阳性表达,主要位于上皮细胞质。推测RBP4基因可能与附睾主细胞分泌功能和附睾头处的收缩功能有关,预示RBP4可能参与附睾微环境的调控,有利于精子成熟、运动和储存。     

Periodic Acid-Schiff Reactive Polysaccharides in Mammalian Spermatozoa During their Epididymal Passage

Contents: The periodic acid-Schiff (PAS) reactive materials of epididymal and ejaculated spermatozoa have been studied in Friesian bulls, buffalo bulls, rams and camels. In the four species, the cytoplasmic droplets displayed strong PAS reaction which did not appear to change markedly during epididymal transit. Only in ram a slight reduction was noted in cauda epididymal and vas deferens spermatozoa. In Friesian bulls, the acrosomes of epididymal spermatozoa exhibited moderate PAS reaction which increased on passage to the vas deferens and after ejaculation. In the buffalo the reaction increased in cauda epididymal and vas deferens spermatozoa but returned again to mderate on ejaculation. In the camel the acrosomes of caput spermatozoa showed strong reaction which was sharply reduced on passage beyond. Meanwhile in rams the acrosomes of spermatozoa obtained from any level of the genital tract exhibited weak PAS reaction. The postnuclear reagions, middle pieces and tails of bull, buffalo, ram and camel spermatozoa showed varying degrees of PAS reaction. The pattern of change during transit of spermatozoa differed from species to another which might be related to the nature of maturational processes. Inhalt: PAS-reaktive Polysaccharide in Saugetierspermien wahrend der Passage durch den Nebenhoden PAS-positive Reaktionen von Nebenhodenspermien und ejakulierten Spermien wurden an Friesian- und Büffelbullen, Schafböcken und Kamelhengsten untersucht. In allen 4 Spezies zeigten die Cytoplasmatröpfchen starke Reaktion, die sich wahrend der Nebenhodenpassage nicht merklich veränderte. Nur beim Schafbock wurde eine geringfügige schwächere Reaktion bei Spermien aus dem Nebenhodenschwanz und dem Vas deferens beobachtet. Bei Friesianbullen zeigte das Akrosom der Nebenhodenspermien eine mittelstarke PAS-Reaktion, die nach Passage durch den Samenleiter und nach der Ejakulation weiter verstarkt war. Beim Büffel war die Reaktion in der Cauda und Vas deferens verstarkt, in ejakulierten Spermien aber wieder schwächer. Beim Kamel war das Akrosom der Caput-Spermien stark positiv, bei der nachfolgenden Passage nahm die Reaktion deutlich ab. Das Akrosom der Schafspermien war in allen Organabschnit-ten nur schwach angefärbt. Bei allen vier untersuchten Spezies zeigten die anderen Spermienbereiche (postnukleare Region, Mittelstück, Schwanz) sehr unterschiedliche Reaktionen. Die Veränderungen während er Nebenhodenpassage waren zwischen den Spezies unterschiedlich und vermutlich abhängzg von Unterschieden in den Reifungsprozessen.     

附睾特异谷胱甘肽过氧化物酶基因调控的研究进展

附睾特异谷胱甘肽过氧化物酶(epididymal specific glutathione peroxidase,GPx5)可保护精子免受氧化应激,维持精子遗传信息的完整性,促进精子发育成熟,其重要生理功能在提高动物繁殖力上有着不可替代的作用,但其基因调控转录表达的机制尚不明确。现作者主要对基因表达特性、表达调控和转录调控等内容进行综述。     

Expression Patterns of T1R1 and T1R3 in the Congjiang Xiang Pig during Epididymal Development

The aim of this experiment was to study the expression pattern of taste receptor family 1 subtypes 1 (T1R1) and 3 (T1R3) during epididymal development of Congjiang Xiang pig, and to explore the possible role of these taste receptors in mammalian male reproductive function and its potential medical value. In this study, the differential expressions of T1R1 and T1R3 in epididymis at 4 key developmental periods (neonatal (15 d), peri-puberty (30 d), puberty (60 d) and sexual maturity (180 d)) of Congjiang Xiang pigs were analyzed. RT-qPCR, immunohistochemistry (IHC) and Western blot were used to detect the changes and distribution of the two taste receptors in epididymis of Congjiang Xiang pigs at different ages. The results of RT-qPCR showed that the expression of TAS1R1 and TAS1R3 mRNA increased gradually from neonatal (15 d) to sexual maturity (180 d), and there was a significant difference between each period (P<0.01). The results of Western blot showed that the expression of T1R1/T1R3 protein was the highest on the 180 d and the lowest on the 15 d. The average protein abundance of T1R1/T1R3 was as follows: 180 d > 30 d > 60 d > 15 d. The results of IHC showed that T1R1 and T1R3 proteins were distributed in the epididymis of Congjiang Xiang pigs at 4 periods, in which T1R1 protein was mainly concentrated in epithelial cell membrane, especially in basal and narrow cells, while T1R3 protein was strongly positive in stereocilia, annular vacuoles and spermatozoa. In summary, the expression of T1R1/T1R3 in the epididymis of Congjiang Xiang pigs increased gradually from 15 d to the peak of sexual maturation, which was related to the differential expression of T1R1/T1R3 in epithelial basal cells, narrow cells and stereocilia of epididymis. These special expression patterns were time related to the physiological function of epididymis, so it is speculated that T1R1/T1R3 are involved in the regulation of sperm maturation and storage in epididymis.     

Differences in aromatase expression and steroid hormone concentrations in the reproductive tissues of male domestic turkeys (Meleagris gallopavo) with white and yellow semen

ABSTRACT

1. To show hormonal differences between male turkeys with yellow semen syndrome (YSS) and white, normal semen (WNS), the expression of aromatase, oestrogen receptor α (ERα), and oestrogen receptor β (ERβ) as well as testosterone and oestradiol concentrations in YSS and WNS testes, epididymis, and ductus deferens were examined.

2. To measure gene expression levels of aromatase and oestrogen receptors (ERs), three complementary techniques (real-time PCR, Western blot, and immunohistochemistry) were used, whereas steroid hormone levels were determined radio-immunologically.

3. Upregulation of aromatase and ERα mRNAs in YSS testes (P < 0.05; P < 0.01), epididymis (P < 0.001; P < 0.001), and ductus deferens (P < 0.05; P < 0.01) compared to those of WNS tissues was detected. Significant increases in the levels of aromatase and ERα proteins were detected in YSS testes (P < 0.001; P < 0.05), epididymis (P < 0.001; P < 0.001), and ductus deferens (P < 0.001; P < 0.05). The expression of ERβ mRNA and protein level was upregulated in the testes (P < 0.05; P < 0.01) and epididymis (P < 0.001; P < 0.01) but not in ductus deferens where it was downregulated (P < 0.01; P < 0.01). Increased intensity of immunoreactive proteins in YSS versus WNS reproductive tissues corroborated gene expression results.

4. Testosterone concentration diminished in YSS epididymis (P < 0.05) and ductus deferens (P < 0.05), but not in the testes, remaining at high level (P < 0.05) compared to WNS values. Concomitantly, increased oestradiol concentration was found in YSS testes (P < 0.05) and epididymis (P < 0.05) but decreased in the ductus deferens (P < 0.05).

5. From the published literature, this study is the first to demonstrate the ability for androgen aromatisation in the turkey reproductive tissues and to show the cellular targets for locally produced oestrogens. The data suggested that the androgen/oestrogen ratio is a mechanistic basis for amplification of differences between turkeys with white and yellow semen and that these results can have a relevance in applied sciences to widen the knowledge on domestic bird reproduction.     

Immunolocalization of some HOX proteins in immature and mature feline testes

Homeobox (HOX) proteins are known for their critical role in body shape formation and tissue differentiation of developing vertebrate embryos. Recent research has shown that HOX proteins have many physiological roles such as cell proliferation, cell cycle, apoptosis and cell differentiation in adults, as well as the development of the vertebrate nerve and reproductive system. This study was conducted to determine the possible physiological functions and expression intensities of HOXA10, HOXA11, HOXB6 and HOXC6 proteins in the male reproductive system (testes, epididymis and deferens ducts), which are important for the continuity of some specific cat breeds in different age ranges. In the study, a total of 18 testicular tissues were used, divided into two groups: less than 6 months (immature) and more than 1 year (mature). Tissue samples were then subjected to immunohistochemical staining with protein-specific antibodies examined in the study. In the findings obtained in the research; it was observed that HOXA10, HOXA11, HOXB6 and HOXC6 produced different intensities of immunolocalization in the epididymis and ductus deferens layers in the immature and mature testicular cells. In addition, it was found that HOXA10 immunoreaction was also seen in some vascular endothelial cells. As a result, it was concluded that the HOX proteins could contribute to the physiological functions of testes, epididymis and ductus deferens and affect male fertility.