Loppion Geraldine, Crespel Amélie, Martinez Anne-Sophie, Auvray Pierrïck, Sourdaine Pascal
UMR M 100 Ifremer, Physiologie et Ecophysiologie des Mollusques Marins, IFR 146 ICORE, Université de Caen Basse-Normandie, Caen, France.
Cell Tissue Res. 2008 Jun;332(3):533-42. doi: 10.1007/s00441-008-0590-z. Epub 2008 Mar 14.
In the lesser-spotted dogfish (Scyliorhinus canicula), spermatogenesis takes place within spermatocysts made up of Sertoli cells associated with stage-synchronized germ cells. As shown in testicular cross sections, cysts radiate in maturational order from the germinative area, where they are formed, to the opposite margin of the testis, where spermiation occurs. In the germinative zone, which is located in a specific area between the tunica albuginea of the testis and the dorsal testicular vessel, individual large spermatogonia are surrounded by elongated somatic cells. The aim of this study has been to define whether these spermatogonia share characteristics with spermatogonial stem cells described in vertebrate and non-vertebrate species. We have studied their ultrastructure and their mitotic activity by 5'-bromo-2'-deoxyuridine (BrdU) incorporation and proliferating cell nuclear antigen (PCNA) immunodetection. Additionally, immunodetection of c-Kit receptor, a marker of differentiating spermatogonia in rodents, and of alpha- and beta-spectrins, as constituents of the spectrosome and the fusome, has been performed. Ultrastructurally, nuclei of stage I spermatogonia present the same mottled aspect in dogfish as undifferentiated spermatogonia nuclei in rodents. Moreover, intercellular bridges are not observed in dogfish spermatogonia, although they are present in stage II spermatogonia. BrdU and PCNA immunodetection underlines their low mitotic activity. The presence of a spectrosome-like structure, a cytological marker of the germline stem cells in Drosophila, has been observed. Our results constitute the first step in the study of spermatogonial stem cells and their niche in the dogfish.
在星点鳐(Scyliorhinus canicula)中,精子发生于由支持细胞与阶段同步的生殖细胞组成的精小囊中。如睾丸横切面所示,精小囊从其形成的生发区开始,按照成熟顺序呈放射状排列至睾丸的相对边缘,即精子释放发生的部位。生发区位于睾丸白膜与睾丸背侧血管之间的特定区域,单个大精原细胞被细长的体细胞所包围。本研究的目的是确定这些精原细胞是否具有脊椎动物和非脊椎动物物种中所描述的精原干细胞的特征。我们通过5'-溴-2'-脱氧尿苷(BrdU)掺入和增殖细胞核抗原(PCNA)免疫检测研究了它们的超微结构和有丝分裂活性。此外,还对啮齿动物中分化精原细胞的标志物c-Kit受体以及作为中心体和融合体组成成分的α-和β-血影蛋白进行了免疫检测。在超微结构上,星点鳐I期精原细胞的细胞核呈现出与啮齿动物未分化精原细胞核相同的斑驳外观。此外,尽管在II期精原细胞中存在细胞间桥,但在星点鳐精原细胞中未观察到。BrdU和PCNA免疫检测突显了它们较低的有丝分裂活性。已观察到一种类似中心体的结构,这是果蝇中生殖系干细胞的细胞学标志物。我们的结果构成了对星点鳐精原干细胞及其微环境研究的第一步。