Expression of small stress protein hsp20 gene in the maturing rat testis

In this study, we cloned a cDNA that encodes a small heat shock protein, Hsp20 (alphaB crystallin-related protein), from a maturing rat testis by means of differential display. The full-length cDNA sequence was completely identical to that registered in the DNA databank. The expression of Hsp20 gene was detected strongly in the heart and slightly in the testis of a 9-week-old rat. The expression of Hsp20 increased gradually from three weeks to 9 weeks, and the strongest expression was observed in the testis at week fifteen. The expression was localized in spermatocytes and round spermatids. The gene expression was not affected by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) when it was administered into male rats during the nursling period.     

Distribution of glycoproteins on feline testicular sperm, epididymal sperm and ejaculated sperm

Glycoproteins (GPs) are known to be involved in the phenomenon of sperm maturation and capacitation. In the present study, we investigated the attachment of GPs on sperm cell membrane during the process of feline sperm maturation from testicular sperm to ejaculated sperm by using 8 FITC-labeled lectins. The results showed that 3 types of GPs were presented on testicular sperm and 7 on caput epididymal sperm. Corpus and cauda epididymal sperm and ejaculated sperm had GPs detected by 8 FITC-labeled lectins used in the present study. This study demonstrates the part of the characteristic of GPs that are present on the feline sperm cell membrane during the process of sperm maturation.     

Effect of a bacterial lipopolysaccharide treatment on rabbit testis and ejaculated sperm

In a previous study, we reported the short- and long-term effects of bacterial lipopolysaccharide (LPS)-induced inflammation on rabbit sperm quality. This study was aimed at exploring the spermatogenesis of the rabbit model focussing on the possible damages occurring to the testis and ejaculated sperm. Twenty New Zealand White rabbit bucks were divided into two groups. One group was inoculated intra-peritoneally with LPS, the other group, considered as control, was treated under the same conditions with saline only. Semen samples were collected before LPS injection, the 7th, 14th, 21st, 30th, 45th, 60th and 90th day after LPS treatment. Semen parameters were evaluated following international guidelines. The kinetic characteristics of ejaculated sperm were analysed using a computer-assisted sperm analyzer and the ultrastructural characteristics were explored by transmission electron microscopy (TEM). On the 7th, 14th and 30th day, testis from treated rabbits and controls were obtained. Testis samples were analysed by light microscopy and TEM. The induced LPS lesions in the testis became evident the 7th day after treatment, with a decrease in germinal cells and with an increase in structurally altered Sertoli cells; normal spermatogenesis was restored on the 30th day. The testicular damages observed on day 7 were probably responsible for the reduction in sperm concentration and motility and the ultrastructural alterations that were detected in the ejaculated sperm on the 14th through the 30th days after treatment. In conclusion, rabbit buck treated with LPS could be a useful model for studying the effect of an induced systemic inflammation on spermatogenesis.     

Distribution and sequence of pyknotic cells in rat fetuses exposed to busulfan

Busulfan, an antineoplastic bifunctional-alkylating agent, is known to induce developmental anomalies. In the present study, we examined the distribution and sequence of pyknotic cells in rat fetal tissues exposed to busulfan. Pregnant rats on gestation day 13 were administered intraperitoneally 30 mg/kg of busulfan, and fetal tissues were examined at 6, 12, 24, 36, 48, 72 and 96 hours after treatment (HAT). Pyknosis of component cells was observed markedly in the brain, moderately in the eyes and spinal cord and mildly in the craniofacial tissue, mandible, limb buds, tail bud, ganglions, alimentary tract, lungs, kidneys, pancreas and liver. In the brain, mitotic inhibition was also detected. Most of the pyknotic cells were considered to be apoptotic cells judging from the results of TUNEL staining and electron microscopic examination. Commonly in the above-mentioned tissues, pyknotic cells began to increase at 24 HAT, peaked at 36 or 48 HAT and disappeared at 96 HAT, which is when the histological picture returned to normal in most tissues except for the brain, spinal cord and eyes. The present study clarified the outline of busulfan-induced apoptosis in rat fetuses.     

精子膜蛋白的研究进展

精子膜蛋白参与了精子获能、顶体反应、精卵融合等生殖过程,具有十分重要的功能.作者就精子膜蛋白在性别控制、免疫不育和避孕、受精前后的作用等3个方面作一综述,并对其将来的发展前景进行了展望.     

Immuno-histological mapping and functional association of seminal proteins in testis and excurrent ducts with sperm function in buffalo

The region-specific expression of seminal proteins in testis and excurrent duct system determines the quality and function of the spermatozoa. In the present study, localization and expression of some of the seminal proteins such as insulin-like growth factor receptor 1β (IGF-1Rβ), phosphatidylethanolamine-binding protein 4 (PEBP4), α-tubulin and tissue factor pathway inhibitor 2 (TFPI2) were carried out in testis, excurrent duct system and spermatozoa of buffalo. IGF-1Rβ was localized in the cells of the seminiferous tubules of the testis, except in primary spermatocytes. The PEBP4 was localized only in the elongated spermatid, whereas α-tubulin and TFPI2 proteins were localized in all cells of the seminiferous tubule including spermatocyte. In the buffalo spermatozoa, IGF-1Rβ, PEBP4, α-tubulin and TFPI2 were localized in the acrosome region, the post-acrosomal region till the tail end, post-acrosome to the entire tail region and the equatorial region, respectively. The study indicates that IGF-1R, α-tubulin and PEBP4 proteins regulate spermatogenesis, whereas TFPI2 may be involved during the zona binding process of the buffalo spermatozoa.     

Immunolocalization of INSL3 in dog foetal Leydig cells and the LGR8 receptor in the gubernaculum testis

Oxidative stress parameters; thiobarbituric acid reaction substances (RBC-TBARS), catalase (RBC-CAT) and reduced glutathione (RBC-GSH)) and the intraerythrocytic concentrations of electrolytes; sodium and potassium (RBC-Na and RBC-K) were determined in 18 well- controlled (WC) and 22 poorly-controlled diabetic mellitus (DM). Dogs with DM had significant higher blood glucose concentration (P < 0.001), haemoglobin A1c (P < 0.01) and fructosamine (P < 0.001) compared to normal healthy dogs (n = 19). Diabetic dogs in both groups had higher RBC-CAT (P < 0.05) while RBC-TBARS were higher significantly only in poorly-controlled DM group (P < 0.05). The RBC-K was significantly higher in both DM groups (P < 0.001). No changes in RBC-GSH and RBC-Na were found between DM and control healthy dogs. By linear regression analysis, the relationship were found between degree of diabetic mellitus and RBC-CAT, RBC-TBARS, RBC-Na and RBC-K. The relationship was also found between oxidative stress parameters and intraerythrocytic K⁺. The results suggest that in diabetic dogs, oxidative stress occurs which related to the severity of disease and may affect potassium homeostasis.     

Increase of methionine aminopeptidase activity in hyperplastic Leydig cells of rat cryptorchid testis: a histochemical study.

Histochemical study on the changes of the aminopeptidase activities in rat testes after surgically-induced cryptorchidism was conducted comparing them with the histochemical changes in regenerated hepatic cells of the partially hepatectomized rat liver. Methionine-aminopeptidase in Leydig cells gradually increased after cryptorchid was induced, whereas the enzyme activity in regenerated hepatic cells decreased. These histochemical observations were coincident with the data obtained by enzyme assay. The present study has indicated that in the rat cryptorchid testis the increase of methionine-aminopeptidase activity was caused by hyperplastic Leydig cells.     

半胱亚磺酸脱羧酶(CSD)基因在大鼠睾丸组织中的表达及其序列分析

试验根据大鼠肝脏牛磺酸生物合成关键酶——半胱亚磺酸脱羧酶(CSD)基因已知核苷酸序列,设计了1对特异的寡核苷酸引物,对提取的睾丸组织总RNA进行RT-PCR扩增,扩增出468 bp的DNA片段,在国际上首次证明CSD基因在大鼠睾丸组织中存在表达,即牛磺酸可在大鼠睾丸组织中生物合成。测定了扩增的睾丸组织CSD基因核苷酸序列,并与已知肝脏CSD基因核苷酸序列及其对应的氨基酸序列作了同源性分析。结果表明:睾丸组织CSD基因核苷酸序列同已知肝脏CSD基因核苷酸序列的同源性为99.8%,其对应氨基酸序列的同源性为100%。     

Changes in interstitial cells during development of buffalo testis

Interstitial cells were identified and counted in the testis of Murrah buffalo calves and bulls at the age of 1, 3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42, 48 and 72 months and older. Six types of cells were identified in the testicular interstitium of 1-month-old calves. These were mesenchymal cells, fetal type Leydig cells, fibroblasts, myoid cells, pericytes and endothelial cells. Adult Leydig cells were visible in 3-month-old calves, but mesenchymal cells were not seen from 18 months onwards. The percentage of mesenchymal cells reached a maximum in 1 month, fetal type Leydig cells in 3 months, adult Leydig cells in 72 months and beyond, fibroblasts in 36 months, myoid cells in 18 months, pericytes in 21 months and endothelial cells after 15 months. Changing percentages of various interstitial cells revealed that myoid cells may have differentiated into fibroblasts and mesenchymal cells, which then differentiated into adult Leydig cells.     

家兔睾丸间质细胞的分离纯化及原代培养

为了研究家兔睾丸间质细胞(Leydigcell,LC)的分离纯化程序及体外生长特点,试验采用4种不同消化程序对家兔睾丸组织进行消化,percoll梯度分离出兔LC,并进行体外培养。结果表明:经4℃保存和34℃孵育,0.5mg/mLⅠ型胶原酶消化40min获得的细胞数最多,经percoll梯度分离的细胞纯度达92%;34℃、5%CO2条件下细胞生长良好,呈现梭形和不规则形2种形状。     

X、Y精子差异膜蛋白及其分离方法的研究进展

X、Y精子差异膜蛋白的分离是利用抗原抗体反应进行性别控制的关键环节,H-Y抗原是最早被发现并应用于性别控制的Y精子特异抗原,但由于抗原特异性较差,未能在生产中得到广泛应用。雄性性别传递偏移、性别偏移及其他相关研究表明,尽管在精子形成过程中,细胞间桥使两类精子部分基因的表达产物被共享了,但X、Y精子膜蛋白差异仍然存在。同时,差异膜蛋白分离技术的进步及这些技术的优化集成,使这种差异膜蛋白的分离成为可能。     

Distribution of the sex chromosome during mouse spermatogenesis in testis tissue sections

During mammalian spermatogenesis, spermatogenic cells undergo mitotic division and are subsequently divided into haploid spermatids by meiotic division, but the dynamics of sex chromosomes during spermatogenesis are unclear in vivo. To gain insight into the distribution of sex chromosomes in the testis, we examined the localization of sex chromosomes before and after meiosis in mouse testis sections. Here, we developed a method of fluorescence in situ hybridization (FISH) using specific probes for the X and Y chromosomes to obtain their positional information in histological testis sections. FISH analysis revealed the sex chromosomal position during spermatogenesis in each stage of seminiferous epithelia and in each spermatogenic cell. In the spermatogonia and leptotene spermatocytes, sex chromosomes were distantly positioned in the cell. In the zygotene and pachytene spermatocytes at prophase I, X and Y chromosomes had a random distribution. After meiosis, the X and Y spermatids were random in every seminiferous epithelium. We also detected aneuploidy of sex chromosomes in spermatogenic cells using our developed FISH analysis. Our results provide further insight into the distribution of sex chromosomes during spermatogenesis, which could help to elucidate a specific difference between X and Y spermatids and sex chromosome-specific behavior.