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1.
Spermatogenesis is a unique developmental sequence involving multiple cell to cell interactions and several categories of regulatory molecules. In contrast to conventional mammalian models in which testicular organization is highly complex, the testis of the dogfish shark Squalus acanthias is technically advantageous for elucidating stage-dependent structural and functional charactericsics and for in vitro regulatory studies. Using incorporation of [3H]thymidine into acid-insoluble molecules as a measure of DNA synthesis by spermatocysts (germ cell/Sertoli cell units) of premeiotic stages, we obtained evidence of a growth inhibitory bioactivity (chalone) within the testis. This activity is differentially distributed (postmeiotic > meiotic > premeiotic), suggesting that more advanced developmental stages, which are upstream in the vascular pathway within the testis, may control the size of the proliferating spermatocyst population and, hence, the advance of less mature stages. These data provide direct evidence for humoral communication between stages of spermatogenesis.  相似文献   

2.
In the dogfish sharkSqualus acanthias different germ cell stages are topographically segregated within the testis. Using this species we have developed methods for the isolation and culture of Sertoli cells from premeiotic, meiotic and post-meiotic stages of spermatogenesis and present preliminary evidence for stage-dependent variations in cell morphology and behavior, thymidine incorporation, protein synthesis and steroidogenesis. The goal of future studies is to determine how maturational changes are regulated in Sertoli cells and, in turn, to elucidate Sertoli cell-germ cell interactions.  相似文献   

3.
The morphometric study of spermatogenic cysts in sexually mature tilapias, during the evolution of spermatogenesis, showed a dramatic increase in both number of germ cells and cyst volume. However, the opposite trend was observed for germ cell size. Nevertheless, the number of Sertoli cells increased gradually up to leptotene/zygotene cysts, stabilizing thereafter. Based on the number of spermatids supported by each Sertoli cell and compared to mammals, Sertoli cell efficiency in tilapias is remarkably high. Sertoli cell proliferation was frequently observed, mainly in spermatogonial cysts, and probably is the major factor related to the testis growth and the increase in sperm production that normally occurs in adult tilapias. The combined duration of spermatocytes (5 days) and spermiogenic (5–6 days) phases of spermatogenesis in fish kept at 25 °C was 10–11 days. Mainly due to acceleration in meiosis, these two phases lasted a total of 6 days in tilapias kept at 30 °C, in the opposite way, at 20 °C spermatogenesis was arrested at pachytene spermatocytes. To our knowledge, this is the most comprehensive investigation performed up to date on testis morphometry and function in adult tilapias.  相似文献   

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SUMMARY: We examined the distribution of two rainbow trout androgen receptors (rtAR: rtAR-α and rtAR-β) in the testis immunohistochemically using a specific antibody to clarify the target cells of androgen in spermatogenesis. Positive rtAR immunoreactivity in paraffin-embedded sections was revealed using microwave treatment, and was detected in the nuclei of Sertoli cells, Leydig cells, and other interstitial cells. The presence of rtAR in Leydig cells suggested that fish androgens regulate Leydig cell activity in an autocrine fashion similar to mammalian androgens. In addition, we found that not all Leydig cells exhibited rtAR immunoreactivity in the mature testis by double staining using anti-3β-hydroxysteroid dehydrogenase (3β-HSD) antibody. Furthermore, rtAR immunoreactivity was also detected in the nuclei of spermatogonia, spermatocytes, and spermatids. The intensity of rtAR immunoreactivity in the nuclei of spermatogonia seemed to be weaker than those of spermatocytes and spermatids. These results suggested that androgens act directly on both germ cells and somatic cells in the regulation of spermatogenesis in the rainbow trout.  相似文献   

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Prohibitin (PHB) is an evolutionarily conserved mitochondrial membrane protein. It plays a vital role in cell proteolysis, senescence, and apoptosis and is associated with spermatogenesis and sperm quality control in mammals. To study the characteristics of the PHB gene and its potential roles during spermatogenesis in Boleophthalmus pectinirostris, we cloned a 1153-bp full-length cDNA from the testis of B. pectinirostris with an open reading frame of 816 bp, which encodes 272 amino acid residues. Real-time quantitative PCR (qPCR) analysis revealed the presence of phb mRNA in all the tissues examined, with higher expression levels found in the testis, kidney, intestine, and muscle tissues. We examined the localization of phb mRNA during spermatogenesis by in situ hybridization (ISH), showing that phb mRNA was distributed in the periphery of the nucleus in primary and secondary spermatocytes. In spermatid and mature sperm, the phb mRNA gradually moved toward one side, where the flagellum is formed. Immunofluorescence (IF) results showed co-localization of the PHB and mitochondria at different stages during spermatogenesis of B. pectinirostris. The signals obtained for PHB decreased as spermatogenesis proceeded; the strongest detection signal was found in secondary spermatocytes, with lower levels of staining in other stages. Additionally, in the mature germ cells, the PHB signals were weak and aggregate in the midpiece of the flagellum.  相似文献   

7.
The gonadotropin (GTH)-stimulated testicular androgen secretionin vitro and the ultrastructure of Leydig and Sertoli cells was studied during the pubertal development in male African catfish. Testicular weight increased from less than 1 mg in the ninth week of age to nearly 600 mg in the 28th week. Immature testes (stage I: spermatogonia) were highly sensitive to GTH and secreted very high amounts of androgens per mg of tissue. The secretion per mg tissue decreased gradually in stages II (spermatogonia and spermatocytes) and III (spermatogonia, spermatocytes, and spermatids), but precipitously in stage IV (all germ cell stages, including spermatozoa). However, due to the testicular weight gain, the total androgen output per pair of testes increased slightly in stage III and strongly in stage IV. The sensitivity to GTH decreased with the appearance of haploid germ cells in stage III. Leydig cells but not Sertoli cells showed the ultrastructural characteristics of steroid producing cells. Leydig cell morphology did not change in stages I–III, while in stage IV, more smooth endoplasmic reticulum was present. The ultrastructural characteristics of Sertoli cells did not change prominently. Thus, spermatogonial multiplication and spermatocyte formation takes place when the testicular steroidogenic system is highly active and responsive to GTH; whereas the differentiation of haploid germ cells is accompanied by a reduced responsiveness to GTH and by the secretion of several-fold lower androgen amounts per mg of tissue.  相似文献   

8.
用兔抗血清对抗促黄体素生成素受体(LHR)或称绒毛膜促性腺激素受体(CGR)和雄激素受体(AR)进行LHR和AR免疫组织化学定位,以揭示外源性促性腺激素(鲤脑垂体激素和hCG)诱发日本鳗鲡精子发生及其内分泌机制。结果表明,经过注射激素处理后的实验组与注射前的对照组相比较,其精巢发育和精子发生出现十分显著的变化。组织学切片观察显示,激素处理前鳗鲡精巢处于精原细胞增殖期,而两种激素混合注射后第10天,实验组可见精小叶中精原细胞的有丝分裂和初级与次级精母细胞的数量显著的增加。注射后第35天,靠近生殖上皮除有少量精原细胞外,精小叶中有大量初级精母细胞和次级精母细胞和少数精子细胞以及管腔中存在少量精子。在注射后第83天,日本鳗鲡完成了精子发生和精巢发育成熟以及释精。免疫组织化学染色结果进一步揭示,激素处理前,LH受体免疫活性分布在生殖上皮,显示强的免疫阳性反应;激素处理后,LH受体定位在Sertoli细胞和间质细胞以及精原细胞和初级与次级精母细胞的胞膜上,均显示强的免疫阳性反应。激素处理前,雄激素受体定位在生殖上皮和早期生精细胞的胞膜上;激素处理后,AR则定位在这些生精细胞的核或胞质,而精子细胞和精子显示免疫阴性反应。这些结果首次证明了这两种激素诱导鳗鲡精子发生和成熟的作用机制是通过LH受体和雄激素受体的介导。  相似文献   

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We conducted histological observation of male germ cells and reproductive organs of the starspotted smooth-hound Mustelus manazo in Tokyo Bay to reveal any abnormality in male reproductive traits, as part of a study to elucidate the factors causing recent fluctuation in abundance of the population. Spermatogenesis proceeded in spermatocysts from the germinal zone in the ventral part of the testis to the degenerative zone in the dorsal part, where the spermatozoa were conveyed into the ciliated lumina of the attached terminal branches of the intratesticular ducts. The intratesticular ducts were classified from their terminal ends into branch, stem, and collecting tubules. The ducts formed in the germinal zone and grew as the spermatocysts developed. An opening formed through the wall of each of the most mature spermatocysts into a branch tubule; bundles of spermatozoa were evacuated through this opening into the branch and then the stem tubule and subsequently into the collecting tubules in the rete testis and the efferent duct connected to the epididymis. Spermatocysts that were unable to emit sperm because of failure of adhesion to the branch tubules were disorganized in situ, as were their spermatozoa. The collapsed spermatocysts seem to be cleared by hemophagocytosis with lymphocytes and leukocytes, which may have been recruited from the epigonal organ. There were no specific abnormalities in the spermatogenesis or the morphological structure of testes, which suggested that an abnormality of male reproductive traits was not the major cause of the recent fluctuation in the population abundance of this species. Details of the intratesticular duct system for sperm emission to the epididymis are the first findings in elasmobranchs worldwide.  相似文献   

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Spermatogenesis and its endocrine regulation   总被引:2,自引:2,他引:2  
Three major phases compose spermatogenesis: mitotic proliferation of spermatogonia, meiosis of spermatocytes, and spermiogenesis, the restructuring of spermatids into flagellated spermatozoa. The process is fuelled by stem cells that, when dividing, either self-renew or produce spermatogonia that are committed to proliferation, meiosis, and spermiogenesis. During all phases, germ cells are in close contact with and require the structural and functional support of Sertoli cells. In contrast to germ cells, these somatic cells express receptors for sex steroids and follicle-stimulating hormone (FSH), the most important hormones that regulate spermatogenesis. A typical Sertoli cell response to an endocrine stimulus would be to change the release of a growth factor that would then mediate the hormone's effect to the germ cells. Recent studies in the Japanese eel have shown, for example, that in the absence of gonadotropin Sertoli cells produce a growth factor (an orthologue of anti-Müllerian hormone) that restricts stem cell divisions to the self-renewal pathway; also estrogens stimulate stem cell renewal divisions but not spermatogonial proliferation. Gonadotropin or 11-ketotestosterone (11-KT) stimulation, however, induces spermatogonial proliferation, which is in part mimicked by another Sertoli cell-derived growth factor (activin B). Since FSH (besides luteinizing hormone, LH) stimulates steroidogenesis in fish, and since FSH is the only gonadotropin detected in the plasma of sexually immature salmonids, increased FSH signalling may be sufficient to initiate spermatogenesis by activating both Sertoli cell functions and 11-KT production. Another important androgen is testosterone (T), which seems to act via feedback mechanisms that can compromise FSH-dependent signalling or steroidogenesis. The testicular production of T and 11-KT therefore needs to be balanced adequately. Further research is required to elucidate in what way(s) 11-KT stimulates later stages of development, such as entry into meiosis and spermiogenesis. At this period, LH becomes increasingly important for the regulation of androgen production. Results from mammalian models suggest that during the later phases, the control of germ cell apoptosis via Sertoli cell factors is an important regulatory mechanism. In many species, sperm cells cannot fertilize eggs until having passed a maturation process known as capacitation, which includes the acquisition of motility. Progestins that are produced under the influence of LH appear to play an important role in this context, which involves the control of the composition of the seminal plasma (e.g., pH values). This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

14.
Spermatogenesis is a highly ordered process in the differentiation of male germ cells. Nuclear morphogenesis is one of the most fundamental cellular transformations to take place during spermatogenesis. These striking transformations from spermatogonia to spermatozoa are a result of phase-specific adaption of the cytoskeleton and its association with molecular motor proteins. KIFC1 is a C-terminal kinesin motor protein that plays an essential role in acrosome formation and nuclear reshaping during spermiogenesis in mammals. To explore its functions during the same process in Larimichthys crocea, we cloned and characterized the cDNA of a mammalian KIFC1 homolog (termed lc-KIFC1) from the total RNA of the testis. The 2481 bp complete lc-KIFC1 cDNA contained a 53 bp 5′ untranslated region, a 535 bp 3′ untranslated region, and a 1893 bp open reading frame that encoded a special protein of 630 amino acids. The predicted lc-KIFC1 protein possesses a divergent tail region, stalk region, and conserved carboxyl motor region. Protein alignment demonstrated that lc-KIFC1 had 73.2, 49.8, 49.3, 54.6, 56.5, 53.1, and 52.1% identity with its homologs in Danio rerio, Eriocheir sinensis, Octopus tankahkeei, Gallus gallus, Xenopus laevis, Mus musculus, and Homo sapiens, respectively. Tissue expression analysis revealed that lc-kifc1 mRNA was mainly expressed in the testis. The trend of lc-kifc1 mRNA expression at different growth stages of the testis showed that the expression increased first and then decreased, in the stage IV of testis, its expression quantity achieved the highest level. In situ hybridization and immunofluorescence results showed that KIFC1 was localized around the nucleus in early spermatids. As spermatid development progressed, the signals increased substantially. These signals peaked and were concentrated at one end of the nucleus when the spermatids began to undergo dramatic changes. In the mature sperm, the signal for KIFC1 gradually became weak and was mainly localized in the tail. In summary, evaluation of the expression pattern for lc-KIFC1 at specific stages of spermiogenesis has shed light on the potential functions of this motor protein in major cytological transformations. In addition, this study may provide a model for researching the molecular mechanisms involved in spermatogenesis in other teleost species, which will lead to a better understanding of the teleost fertilization process.  相似文献   

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为探究Smoothened (Smo)信号在精巢不同细胞增殖与存活中的作用,实验分离鉴定了尼罗罗非鱼smo (命名为Onsmo),检测了其在不同组织中的表达分布及在精巢中的细胞表达模式,在尼罗罗非鱼精巢组织的体外培养体系中,用Smo特异性激动剂SAG或抑制剂环巴胺分别进行处理,EdU掺入法及TUNEL法检测了处理后生殖细胞(Vasa+)、Sertoli细胞(Amh+)与Leydig细胞(Cyp17a1+)增殖或凋亡情况。结果显示,Onsmo开放阅读框全长2 478 bp,编码825个氨基酸,含有7次跨膜结构域,与人SMO氨基酸一致性达77%;Onsmo表达于包括精巢在内的多个组织;在精巢中,Onsmo在多种不同类型细胞表达,包括精原细胞、精母细胞、Sertoli细胞以及Leydig细胞;在精巢组织的体外培养体系中,SAG处理对精原细胞增殖具有显著促进作用,而环巴胺处理对Sertoli细胞、Leydig细胞凋亡具有显著促进作用。研究表明,On Smo信号在尼罗罗非鱼精巢精原细胞增殖与体细胞存活中具有重要作用。该研究首次证实...  相似文献   

18.
In fish, sex steroids initiate and/or accelerate the maturation of the brain-pituitary-gonad axis. In order to obtain information on the steroid milieu during the pubertal development of male African catfish, we have monitored the conversion of [3H]-pregnenolone and [3H]-androstenedione by testis and [3H]-pregnenolone by interrenal tissue fragmentsin vitro. Pubertal development occurs between two and six months of age. Testicular development proceeds through four main stages that are characterised histologically by the presence of spermatogonia (stage I), spermatogonia and spermatocytes (stage II), spermatogonia, spermatocytes and spermatids (stage III), and all germ cells including spermatozoa (stage IV). 11β-Hydroxyandrostenedione and cortisol were the main products of testes and interrenal tissue, respectively, in all stages of the pubertal development; a change in the steroidogenic pattern was not observed during this period. The interrenal tissue displayed no significant conversion of [3H]-pregnenolone to 11-oxygenated androgens. Blood plasma was analyzed for the presence of five androgens; testosterone, 11β-hydroxytestosterone, 11β-hydroxyandrostenedione, androstenetrione, and 11-ketotestosterone. 11-Ketotestosterone was the quantitatively dominating androgen in the circulation at all stages of development, which was more pronounced in stages III and IV. The obvious differences between thein vitro andin vivo results, namely 11β-hydroxyandrostenedione being the main testicular productvs. 11-ketotestosterone dominating in the blood, may indicate that 11β-hydroxyandrostenedione is converted to 11-ketotestosterone at extratesticular sites.  相似文献   

19.
为检验SCP3在无脊椎动物中是否可以标记特定的生殖细胞,实验利用RACE技术克隆了栉孔扇贝SCP3(Cf-SCP3)的全长cDNA序列,结果显示,其长度为1 033 bp,开放阅读框为726 bp,编码241个氨基酸。推导的氨基酸序列中包含SCP3保守的Cor1结构域和卷曲螺旋结构域。制备了Cf-SCP3地高辛标记的RNA探针和Cf-SCP3重组蛋白的多克隆抗体,采用原位杂交和免疫组织化学技术检测其细胞学定位,结果显示Cf-SCP3转录本和Cf-SCP3蛋白均特异性地定位在栉孔扇贝的初级精母细胞和未受精卵中,在精巢的其他类型细胞和卵巢中均未见表达信号。研究表明,Cf-SCP3蛋白可能与脊椎动物的SCP3蛋白一样,作为联会复合体的组成成分参与栉孔扇贝的配子发生,同时可以作为一个栉孔扇贝初级精母细胞的分子标记应用到体外诱导生殖细胞分化的研究。  相似文献   

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为探索龟鳖类生殖细胞的发育分化机制,实验通过特异性引物克隆了中华鳖dazl基因的cDNA片段,长1 007 bp,其中包括5′端非编码区197 bp,3′端非编码区33 bp,开放阅读框777 bp,编码258个氨基酸。氨基酸序列比对显示其与西部锦龟Dazl同源性最高,达96%;与小鼠Dazl同源性达75%。荧光定量PCR分析结果显示,中华鳖dazl mRNA主要在精巢和卵巢中表达,在体细胞组织中仅检测到微量表达。冰冻切片原位杂交结果显示,中华鳖dazl mRNA在生殖细胞中特异表达,且在不同时期的生殖细胞中呈动态表达模式。在精巢中,中华鳖dazl mRNA在初级和次级精母细胞中表达最强,在精原干细胞中表达水平次之,在精子细胞中未检测到信号;在卵巢中,中华鳖dazl mRNA信号在初级卵母细胞胞质中均匀分布且在最早期的初级卵母细胞中信号最强,随着卵母细胞的增大,信号逐渐聚集并逐渐减弱。研究表明,dazl基因可能对中华鳖两性生殖细胞的发生具有重要的调控作用。  相似文献   

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