Morphological evaluation of spermatogenesis in Lake Magadi tilapia (Alcolapia grahami): a fish living on the edge.

Spermatogenesis in Lake Magadi tilapia (Alcolapia grahami), a cichlid fish endemic to the highly alkaline and saline Lake Magadi in Kenya, was evaluated using light and transmission electron microscopy. Spermatogenesis, typified by its three major phases (spermatocytogenesis, meiosis and spermiogenesis), was demonstrated by the presence of maturational spermatogenic cells namely spermatogonia, spermatocytes, spermatids and spermatozoa. Primary spermatogonia, the largest of all the germ cells, underwent a series of mitotic divisions producing primary spermatocytes, which then entered two consecutive meiotic divisions to produce secondary spermatocytes and spermatids. Spermatids, in turn, passed through three structurally distinct developmental stages typical of type-I spermiogenesis to yield typical primitive anacrosomal spermatozoa of the externally fertilizing type (aquasperm). The spermatozoon of this fish exhibited a spheroidal head with the nucleus containing highly electron-dense chromatin globules, a midpiece containing ten ovoid mitochondria arranged in two rows and a flagellum formed by the typical 9 + 2 microtubule axoneme. In addition, the midpiece, with no cytoplasmic sheath, appeared to end blindly distally in a lobe-like pattern around the flagellum; a feature that was unique and considered adaptive for the spermatozoon of this species to the harsh external environment. These observations show that the testis of A. grahami often undergoes active spermatogenesis despite the harsh environmental conditions to which it is exposed on a daily basis within the lake. Further, the spermiogenic features and spermatozoal ultrastructure appear to be characteristic of Cichlidae and, therefore, may be of phylogenetic significance.

Khat (Catha edulis) lowers plasma luteinizing hormone (LH) and testosterone secretion, but increases cortisol levels in male rabbits.

AIM: This study investigated the effects of fresh khat extract on specific circulating hormones in male rabbits. MATERIALS AND METHODS: A total of 25 male New Zealand White rabbits were divided into five groups each comprising five animals. The first four groups were fed four doses (1.5 g/kg, 4.5 g/kg, 13.5 g/kg and 40.5 g/kg body weight) of khat extract twice a week for 5 weeks while the last group, serving as control, was fed only normal saline via intragastric tube. Blood samples were collected at 15 min interval for up to 3 h after khat extract administration and plasma assayed for luteinizing hormone (LH), testosterone and cortisol levels using radioimmunoassay technique. RESULTS: Khat extract at all doses significantly lowered (P<0.05) LH pulse frequency, area under LH curve, mean plasma LH and mean plasma testosterone levels. Plasma cortisol levels were significantly elevated (P<0.05) in khat-treated rabbits in a dose-dependent manner. CONCLUSION: This study demonstrates that khat may impair reproductive function in male rabbits by interfering with sex hormone profiles.

Early testicular response to intraperitoneal administration of ethane dimethanesulphonate (EDS) in the goat (Capra hircus).

Early morphological changes in the goat testis after a single intraperitoneal injection of ethane dimethanesulphonate (EDS) were investigated using both light and electron microscopy. The compound was administered at two dose levels: 75 mg/kg and 25 mg/kg. While the former resulted in some deaths due to toxicity, the latter had no noticeable toxic effects on the animals. The testicular effects at both dose levels were similar. Six (6) days post-treatment, Leydig cells were refractory to EDS challenge but there was a marked disruption of spermatogenesis. These Leydig cells exhibited ovoid or irregularly round nuclei, abundant cytoplasm containing spherical, ovoid or elongate mitochondria and a preponderance of smooth endoplasmic reticulum typical of the normal cells. Lipid droplets were rare. In the seminiferous tubules germ and Sertoli cell degeneration was observed. Changes in the germ cells included: spermatogonial degeneration, condensed chromatin in leptotene spermatocytes and failure of chromatin re-organization resulting in the formation of clumps in the cells at the telophase stage of cell division (stage 4 of the seminiferous cycle). The nuclear envelope of primary spermatocytes showed marked irregularity and there was an overall reduction in cell size. There was peripheral re-distribution of chromatin in developing spermatids of stages 1, 2 and 5, often resulting in thick margination along the nucleolemma and leaving a pale nucleoplasm. An accompanying retention of maturation phase spermatids in stage 2 tubules was also observed. Sertoli cells exhibited extensive accumulation of intracytoplasmic vesicles, obscuring the rest of the organelles. Intercellular vacuoles also occurred within the epithelium. The results suggest that while EDS does not have any effect on goat Leydig cells, it causes severe disruption of the spermatogenic process. Furthermore, it is concluded from the results that the optimum dose in this species is 25 mg/kg.

Epididymal epithelial cell involution following a single intraperitoneal administration of ethane dimethanesulfonate in the goat (Capra hircus).

Ethane dimethanesulfonate (EDS) selectively destroys Leydig cells in rats and a few other smaller animal species but not in mice and quail. In the teleost fish, it stimulates testicular activity instead. It also causes formation of sperm granulomas, reduction of sperm fertilizing ability, and destruction of clear cells in the epididymis. Investigations involving larger animal species are scanty. We have previously reported that EDS has no effect on goat Leydig cells but appears to have a direct cytotoxic effect on the seminiferous epithelium. This study was therefore designed to investigate the effects of EDS on goat epididymal cytoarchitecture. EDS was administered intraperitoneally at two dose levels, 75 and 25 mg/kg body wt. The former dose was rather toxic, killing three of five goats in this group within 24 h whereas the latter dose was well tolerated. Six days after treatment, the goats were hemicastrated and the epididymis was isolated and processed for light and electron microscopy. Involution associated with EDS was observed in epithelial cells of all regions of the epididymis, each having its own specific and peculiar changes. In the caput, there was increased cytoplasmic density accompanied by enlarged vacuoles and paucity of secretory vesicles in the apical cytoplasm. The Golgi cisternae were dilated and disorganized and, in the basal aspect, large dense staining bodies or inclusions, degenerative mitochondria, and lamellated bodies were observed. In the corpus, large vacuoles containing flocculent materials occurred in the entire cell cytoplasm but were particularly numerous and large in the midcytoplasm, completely obliterating the Golgi area. There was a dramatic reduction in epithelial height in the cauda epididymis accompanied by sparse distribution of markedly shortened microvilli. The epithelial cells had extensively lobulated nuclei and disorganized cytoplasm with dilated Golgi apparatus and large conglomerations of tubular structures. These structural changes suggest that EDS causes degeneration of goat epididymal epithelial cells. These effects are likely to result from the direct action of the compound on the epithelium.

Morphological characterization of the seminiferous cycle in the goat (Capra hircus): a histological and ultrastructural study.

The cycle of spermatogenesis/seminiferous cycle was investigated in the goat testis using both light and electron microscopy techniques. Using the various cell associations and the accompanying changes in spermatid shape and location, the cycle was divided into eight (8) successive stages. The cycle began with the accomplishment of spermiation (stage 1) and ended with apical migration and close attachment of late maturation phase spermatids at the Sertoli cell apex accompanied by adluminal retention of residual bodies with dense staining inclusions (stage 8). The early stages of the cycle (stages 1-4) were therefore characterized by the presence of only one generation of spermatids, the second one appearing only after the division of secondary spermatocytes in stage 4. Consequently, stages 5-8 had two generations of spermatids; Golgi or cap phase as well as maturation phase spermatids. Although stages 5 to 7 appeared as distinct entities, stages 6 and 7 were rather short-lived and considered as continuations of stage 5. Therefore, the 8 stages of the cycle in the goat were further condensed into 6 main divisions. The duration of each stage was estimated by the frequency of occurrence in sections. Among these, stage 1 had the highest frequency (34%) followed by stages 5-7 (27%). Stages 8 and 4 had the shortest frequency (up to 9%) while stages 2 and 3 had 13% and 12% respectively. These results indicate that, like most domestic species, goats have a cycle of 8 stages with 6 main divisions, the longest being stage 1.

In vitro production of testosterone and plasma levels of luteinising hormone, testosterone and cortisol in male rats treated with heptachlor.

Male rats were divided into six groups of five rats each. Rats were injected subcutaneously with different concentrations of heptachlor for 2 weeks. Heptachlor at all doses significantly suppressed plasma testosterone levels (P < 0.05). Plasma luteinizing hormone (LH) (P < 0.01) and cortisol (P < 0.02) levels were significantly elevated in heptachlor-treated rats as compared to corn oil-treated controls. LH and testosterone levels showed strong correlation (r = 0.69, P < 0.05). The testes in rats treated with 25 mg/kg body weight of heptachlor showed some pathological changes. We conclude that heptachlor causes adverse effects on several male reproductive parameters in rats.

Ultrastructural study of the testis of non-breeding naked mole-rat (Heterocephalus glaber, Ruppell).

The testicular structure of the wild caught naked mole rat was studied. It comprises of a large volume of lipid-rich interstitial cells of Leydig among which are few scattered seminiferous tubules. In addition, the interstitial cells possess elongated mitochondria and vast network of smooth endoplasmic reticulum (sER). The Golgi apparatus (GA) apparently is not conspicuous or well developed. All stages of spermatogenesis occur in the seminiferous tubules although the mature forms (secondary spermatocytes, spermatids and spermatozoa) are few. Sertoli cells show an irregular nucleus, mitochondria oriented perpendicular to the basement membrane, a vast network of endoplasmic reticulum with sER as the predominant form and lipid droplets. The ultrastructural features of Leydig cells seem to suggest a steroidogenic capacity although the vast accumulation of lipid droplets may imply impaired utilisation of cholesterol reservoir as a result of pituitary hormonal imbalance or (and) the local paracrine influence by Sertoli cells. The cause of slow-down in spermatogenesis is still unclear but may also be under the influence of pheromonal cues or the local paracrine control. Sertoli cell features point towards a role of synthesis and secretion.