Gonadal characterization of the Amazonian fish Serrapinnus kriegi (Characidae: Cheirodontinae)

The knowledge of the testicular and ovarian morphology of a particular fish species is of paramount importance. Such analyze enables the development of studies and techniques aiming the improvement of their reproduction, management, commercialization and even their conservation. This study performed the ovarian and testicular characterization of the ornamental Amazon fish Serrapinnus kriegi. A total of three males and three females had their gonads analyzed by optical microscopy. Females present ovaries filled with oocytes in asynchronous development, indicating partial spawning in the species. Moreover, the micropyle and micropilar cell formation was observed in primary growing oocytes, representing a precocious oocyte development; and the zona radiata in the final vitellogenic oocytes is thicker than other related species, evidencing the development of a better protection to the embryos in function of the waters’ turbulence that characterize it spawning sites in the Amazonian streams. The male specimens’ present anastomosed tubular testes with unrestricted spermatogonia spread along the entire seminiferous tubules. The present data elucidate the dynamic of spermatogenesis and oogenesis of an ornamental Amazonian species, through the description of the male and female germ cells development.

Acute Exposure to the Neonicotinoid Insecticide Imidacloprid of Zebrafish (Danio rerio) Gonads: A Histopathological Approach

Neonicotinoids are the new class of insecticides that are high target specificity to insects. Imidacloprid is a neonicotinoid that is the most widely used insecticide in the world. As a result of its widespread use in agriculture, imidacloprid interferes with the aquatic system and threatens the aquatic environment. In this study, an investigation of the histopathological effects of imidacloprid on zebrafish gonads was aimed. Zebrafish were exposed to 9.5 mg/L, 19 mg/L, and 38 mg/L of imidacloprid for 5 days, considering the 96 h LC50 value. After dissecting the gonadal tissues, routine histological techniques were applied, and the tissues were stained with Periodic Acid-Schiff (PAS), Toluidine Blue (TB), and Haematoxylin and Eosin (H&E). Sections were examined under a light microscope. While normal gonad histology was observed in the control group, histopathological alternations such as degeneration and union in the seminiferous tubules, hypertrophy in spermatogenic and Leydig cells, and interstitial fibrosis were detected in testicular tissue of the experimental groups. In the ovarian tissues of the experimental groups, structural deterioration in oocytes, autolysis, increase in the number of atretic oocytes, vacuolization in cortical alveoli, thickening and curling in the zona radiata, and opening in the perifollicular layers were detected.

Histopathological and apoptotic examination of zebrafish (Danio rerio) gonads exposed to triclosan

Triclosan, produced as a broad-spectrum antibiotic in the early 1960s, is generally used as a preservative in personal care products, fabrics, plastic products such as kitchenware and toys. As a result of the high demand for triclosan, this chemical threatens the aquatic ecosystem by contaminating wastewater sources. Environmental pollutants affect the reproductive potential of fish, one of the most critical aquatic organisms. This study aimed to investigate the histopathological and apoptotic effects of triclosan in zebrafish gonads. Fish were exposed to sublethal concentrations of triclosan for 5 days, and general histological methods were applied. Histological sections were examined under a light microscope after staining with hematoxylin and eosin and toluidine blue. Triclosan exposure caused deterioration in ovarian tissue, such as shrinkage in the ooplasm, accumulation of proteinaceous fluid in the interstitial tissue, morphological changes of oocyte and the zona radiata. In testicular tissue, triclosan exposure caused fusion in seminiferous tubules, hypertrophy in spermatogenic and Leydig cells, edema in seminiferous tubules, and karyorrhexis in spermatogenic cells. The TUNEL assay was used for the determination of apoptotic cells. Brown-colored apoptotic cells were visualized under the light microscope. TUNEL positive cells were observed in all exposure groups. Triclosan administration was found to cause apoptosis in zebrafish gonads. These findings indicate that triclosan potentially affects fish reproduction, and that its judicious disposal is essential for protecting the environment and maintaining the reproductive potential of fish.

Knockout of the Glucocorticoid Receptor Impairs Reproduction in Female Zebrafish

The pleiotropic effects of glucocorticoids in metabolic, developmental, immune and stress response processes have been extensively investigated; conversely, their roles in reproduction are still less documented. It is well known that stress or long-lasting therapies can cause a strong increase in these hormones, negatively affecting reproduction. Moreover, the need of glucocorticoid (GC) homeostatic levels is highlighted by the reduced fertility reported in the zebrafish glucocorticoid receptor mutant (nr3c1ia30/ia30) line (hereafter named gr−/−). Starting from such evidence, in this study, we have investigated the role of glucocorticoid receptor (Gr) in the reproduction of female zebrafish. Key signals orchestrating the reproductive process at the brain, liver, and ovarian levels were analyzed using a multidisciplinary approach. An impairment of the kiss-GnRH system was observed at the central level in (gr−/−) mutants as compared to wild-type (wt) females while, in the liver, vitellogenin (vtg) mRNA transcription was not affected. Changes were instead observed in the ovary, particularly in maturing and fully grown follicles (classes III and IV), as documented by the mRNA levels of signals involved in oocyte maturation and ovulation. Follicles isolated from gr−/− females displayed a decreased level of signals involved in the acquisition of competence and maturation, causing a reduction in ovulation with respect to wt females. Fourier transform infrared imaging (FTIRI) analysis of gr−/− follicle cytoplasm showed major changes in macromolecule abundance and distribution with a clear alteration of oocyte composition. Finally, differences in the molecular structure of the zona radiata layer of gr−/− follicles are likely to contribute to the reduced fertilization rate observed in mutants.

Gonadal development and gametogenesis in the threatened freshwater siluroid fish, Horabagrus brachysoma (Gunther, 1864): A morpho-histological perspective

The macro-morphology and histology of the reproductive system together with cytology of the process of gametogenesis in Asian sun catfish, Horabagrus brachysoma (Gunther, 1864) were investigated. A maturity key for horabagrids, based on macroscopic appearance of gonads at different stages of maturity was established for the first time. The distinctive features of male genital system were the digitiform testes, absence of seminal vesicles and five morphological maturity stages. Unrestricted semicystic spermatogenic activity with seven cytological stages was recognised viz., primordial germ cells, primary spermatogonia, secondary spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids and spermatozoa. The classic cystovarian, group synchronic ovary, with five morphological maturity stages and a single spawning period were the peculiarities of female reproduction. The process of oogenesis progressed through five phases viz., previtellogenic, cytoplasmic growth, vitellogenic, yolky and mature. The sequence of oocyte maturation was: oogonia, chromatin nuclear, peri-nuclear, cortical alveolar, yolky and mature. Vitellogenesis was initiated in oocytes of maturing or stage-Il ovary. No liqueification of yolk was observed. Atresia was observed in vitellogenic and mature oocytes only. Corpus luteum like structure was present in ovaries with vitellogenic and yolky oocytes. A very prominent zona radiata of thickness 16.5 μm covered mature oocytes.

Histochemical and morphological features of biopsied and stripped oocytes from the Brazilian endangered teleost pirapitinga, Brycon nattereri (Characiformes)

SummaryThe aim of the present study was to characterize biopsied and stripped oocytes from the Brazilian endangered teleost pirapitinga (Brycon nattereri) using histochemical and morphological analyses. Biopsied oocytes had a mean diameter of 2.225 mm (modal diameter: 2.312 mm), complete vitellogenesis and a central or slightly eccentric nucleus. Neutral polysaccharides were detected in the follicular cells, zona radiata and yolk globules, while acidic polysaccharides were detected in the follicular cells and cortical alveoli. Ten out of the 19 females treated with two doses of carp pituitary extract (cPE) released oocytes, which were also analysed. Stripping occurred 292 ± 39 degree-hours after the second dose of cPE and led to a mean spawning weight of 36.2 g, 10% spawning index, 241 oocytes/g of ova, 8222 oocytes/female and 23 oocytes/g of body weight. Stripped oocytes had a mean diameter of 2.33 mm and a mode at 2.375 mm, were weakly adhesive and coloration ranged from wine to brown. Under scanning electron microscopy, stripped oocytes exhibited a single funnel-shaped micropyle located at the animal pole and a zona radiata that measured 7.7 μm in thickness with eight pore canals/μm2. Oocyte morphology in Brycon nattereri is similar to that found in other species of the genus, except for the larger size and weaker adhesiveness. These findings provide essential information for a better understanding of the reproductive biology of B. nattereri and the establishment of conservation measures for this threatened species.