Characterization of undifferentiated spermatogonia and the spermatogonial niche in the lambari fish Astyanax altiparanae.

Undifferentiated type A spermatogonia are the foundation of fish spermatogenesis. This cell population includes the spermatogonial stem cell population, which is able to either self-renew or differentiate into cells that will generate the male gamete - the spermatozoa. Spermatogonia stem cells are located in a specific region of the testes known as the spermatogonial niche, which regulates spermatogonial stem cell function. This study characterizes undifferentiated type A spermatogonia and their S-phase label-retaining cell properties in Astyanax altiparanae testes. This is a fish species of substantial commercial, environmental, and academic importance. Two types of undifferentiated spermatogonia have been described in A. altiparanae testis: Aund* and Aund. Among the main differences observed, Aund* spermatogonia have an irregular nuclear envelope, decondensed chromatin, one or two nucleoli, and nuages in the cytoplasm; meanwhile, type Aund have a round nucleus. Aund* is preferentially distributed in areas neighboring the interstitial compartment, whereas Aund is located in the intertubular area. Finally, this study found that undifferentiated type A spermatogonia were able to retain BrdU over a long chase period, suggesting that these cells have a long cell cycle and potential stem cell candidates among them. Based on these findings, undifferentiated type A spermatogonia may be characterized as putative stem cells in A. altiparanae testis. This work will contribute to further studies on the stem cell biology of this promising Neotropical experimental model.

Involution processes of follicular atresia and post-ovulatory complex in a characid fish ovary: a study of apoptosis and autophagy pathways.

Recent studies seem to indicate that apoptosis and autophagy can act cooperatively in fish ovaries in order to achieve more effective ovarian regression after spawning. Considering the importance of tissue remodeling in ovarian functioning, we sought to morphologically characterize the involution processes of follicular atresia and post-ovulatory complexes using Astyanax altiparanae as an experimental model, and to determine the location of proteins involved in apoptosis and autophagy throughout this process. Fifteen females were collected after reproductive management. Fragments of the left ovaries were removed, fixed, and prepared for light microscopy and immunofluorescence analyses. The main characteristics of the involution processes were found to be consistent with previous descriptions. However, there were certain morphological peculiarities that do not appear to have been described for any other species thus far. These peculiarities may be related to the focus of this study on a single species, which allows for a more detailed investigation into morphological changes than studies on multiple species. Autophagy was also found to precede apoptosis in both involution processes in A. altiparanae. This may be related to the energy recycling process required before the removal of degenerated follicular cells by apoptosis. Thus, these results support the idea that there is crosstalk between autophagy and apoptosis pathways in ovarian involution processes, as well as the idea that the cell death pathways of these processes are conserved between teleost species with external fertilization.

Spermatogonia, Germline Cells, and Testicular Organization in the Characiform Prochilodus lineatus Studied Using Histological, Stereological, and Morphometric Approaches.

Prochilodus lineatus is an important representative of the order Characiformes and a species that offers great advantages to fish farming. Therefore, detailed knowledge of its reproductive biology can be applied to various fields of production and biotechnology. In this study, we have identified testicular germ cells during spermatogenesis and have evaluated the volumetric proportion of the testes occupied by structures of the tubular and intertubular compartments. In addition, the individual volume of type A spermatogonia was measured and used to estimate the mean number of these cells per testis. Gonads of adult P. lineatus males were extracted and fixed. Light and transmission electron microscopy were applied to fragments of three testicular regions. Histological, stereological, and morphometric analyses were performed. The stereological data suggest that components of the tubular and intertubular compartments of the P. lineatus testes present a uniform distribution in all three regions and therefore reflect regions with similar distributions of cell types. In addition, P. lineatus testes showed ∼0.6% of type A spermatogonia, as well as a predominance of cysts of primary spermatocytes and spermatids during the reproductive phase evaluated. The results from this study provide a better understanding of the morphology and structure of the testis and of the characterization of the type A spermatogonia in P. lineatus. The nuclear diameter of germ cells also decreases significantly during spermatogenesis. The data presented herein are the first of its kind for the order Characiformes and may be useful for future biotechnology studies on fish reproduction. Anat Rec, 300:589-599, 2017. © 2016 Wiley Periodicals, Inc.