Germ cell transplantation as a potential biotechnological approach to fish reproduction.

Although the use of germ cell transplantation has been relatively well established in mammals, the technique has only been adapted for use in fish after entering the 2000s. During the last decade, several different approaches have been developed for germ cell transplantation in fish using recipients of various ages and life stages, such as blastula-stage embryos, newly hatched larvae and sexually mature specimens. As germ cells can develop into live organisms through maturation and fertilization processes, germ cell transplantation in fish has opened up new avenues of research in reproductive biotechnology and aquaculture. For instance, the use of xenotransplantation in fish has lead to advances in the conservation of endangered species and the production of commercially valuable fish using surrogated recipients. Further, this could also facilitate the engineering of transgenic fish. However, as is the case with mammals, knowledge regarding the basic biology and physiology of germline stem cells in fish remains incomplete, imposing a considerable limitation on the application of germ cell transplantation in fish. Furthering our understanding of germline stem cells would contribute significantly to advances regarding germ cell transplantation in fish.

Androgen receptor signalling in peritubular myoid cells is essential for normal differentiation and function of adult Leydig cells.

Testosterone synthesis depends on normal Leydig cell (LC) development, but the mechanisms controlling this development remain unclear. We recently demonstrated that androgen receptor (AR) ablation from a proportion of testicular peritubular myoid cells (PTM-ARKO) did not affect LC number, but resulted in compensated LC failure. The current study extends these investigations, demonstrating that PTM AR signalling is important for normal development, ultrastructure and function of adult LCs. Notably, mRNAs for LC markers [e.g. steroidogenic factor 1 (Nr5a1), insulin-like growth factor (Igf-1) and insulin-like factor 3 (Insl3)] were significantly reduced in adult PTM-ARKOs, but not all LCs were similarly affected. Two LC sub-populations were identified, one apparently 'normal' sub-population that expressed adult LC markers and steroidogenic enzymes as in controls, and another 'abnormal' sub-population that had arrested development and only weakly expressed INSL3, luteinizing hormone receptor, and several steroidogenic enzymes. Furthermore, unlike 'normal' LCs in PTM-ARKOs, the 'abnormal' LCs did not involute as expected in response to exogenous testosterone. Differential function of these LC sub-populations is likely to mean that the 'normal' LCs work harder to compensate for the 'abnormal' LCs to maintain normal serum testosterone. These findings reveal new paracrine mechanisms underlying adult LC development, which can be further investigated using PTM-ARKOs.

Duration of spermatogenesis in the bullfrog (Lithobates catesbeianus).

The bullfrog (Lithobates catesbeianus) has substantial economic importance and has also been used as an experimental model for biological studies in the fields of pharmacology, medicine, and reproductive biology, especially studies addressing gametogenesis. However, there is a lack of comprehensive information in the literature regarding testis structure and function in this amphibian. The main objective of the current study was to estimate the duration of the various phases of spermatogenesis in this vertebrate. Sixteen sexually mature bullfrogs received an intracoelomic administration of tritiated thymidine. Testes were analyzed at various times between 1h and 33 d after administration to detect the most advanced germ cell types labeled at each interval, as well as labeled preleptotene spermatocytes, which presumably originated from spermatogonial stem cells. The duration of the spermatogonial, spermatocytic, and spermiogenic phases of spermatogenesis in the bullfrog were approximately 18, 14, and 8 d, respectively. Thus, the total duration of the spermatogenesis process from early spermatogonia through to spermatozoa was 40 d in this species, similar to that of most previously investigated mammalian species. To our knowledge, this is the first reliable report on the duration of the full spermatogenic process in any amphibian species. These findings will be very useful for tracking the pace of germ cells in studies involving spermatogonial transplantation in lower vertebrates.

Morphologic changes in the urethral epithelium in an ethanol-drinking rat strain (UChA and UChB).

The extreme use of ethanol causes metabolic and pathologic changes in testes and urogenital system in different animal species. The enzyme alcohol dehydrogenase (ADH) catalyses the conversion of ethanol into carcinogenic metabolite acetaldehyde which is partly excreted into the urine. However, papers relating the chronic ethanol consumption to the urethral morphology are unknown. This work evaluates the toxic effect of the chronic ethanol ingestion on the urethral epithelium of UChA and UChB rats. Conventional techniques of histology, histochemistry, immunohistochemistry and ultrastructural analysis were used. The analysis showed the presence of lipid drops and intercellular spaces in the epithelial cells in the urethra of UChA and UChB rats compared to control rats. Urethral neuroendocrine cell were observed and characterized for presenting vesicles containing electron-dense granules associated with nervous fibers. We conclude that the chronic consumption of ethanol induces the presence lipid drops in the epithelial cells of the urethra of UChA and UChB rats. The NE cells of the urethra of UChA and UChB rats did not show alterations under chronic effect of the ethanol.

Morphological alterations on the prostate of Calomys callosus submitted to chronic ethanol ingestion.

The objective of the present study was to assess the possible toxic effects of chronic alcohol ingestion on the ultrastructure of the glandular epithelium of the prostate of the rodent Calomys callosus, in order to contribute to the understanding of the consequences of alcohol abuse for the morphology of the male reproductive apparatus. Sixteen adult animals aged three months were divided into two experimental groups. The control group received a solid diet and tap water, and the alcoholic group received the same solid diet and ethanol P.A. diluted 20% in water (v/v). After 120 days of treatment, all animals were anesthetized, weighed and sacrificed. At the end of treatment, mean body weight did not differ between control and alcoholic animals. The prostate epithelial cells of the alcoholic group showed intense atrophy and ultrastructural alterations such as the presence of lipid droplets, altered nuclei, ruptured mitochondrial cristae, and intense dilatation of the cisterns of the granular endoplasmic reticulum. It was concluded that 20% ethanol provokes marked lesions on the epithelium of the prostate probably interfering on the glandular secretion.

Ultrastructural changes on the hard palatine mucosa of Calomys callosus after 120 days of experimental chronic alcoholism.

The present study analyzed the toxic effects of chronic alcohol ingestion on the ultrastructure of the lining epithelium of the hard palatine mucosa of the rodent Calomys callosus, in order to contribute to the understanding of the consequences of alcohol abuse for the morphology of the digestive system. Twenty-six adult animals aged three months were divided into two experimental groups. The control group received a solid diet and tap water, and the alcoholic group received the same solid diet and ethanol P.A. diluted 20% in water (v/v). After 120 days of treatment, all animals were anaesthetised, weighed and sacrificed. At the end of treatment, mean body weight did not differ between control and alcoholic animals. The epithelial cells of the alcoholic group showed many alterations such as the presence of lipid droplets, nuclei in corneum layer, nuclei with increase peripheral chromatin and greater electron density, altered mitochondria, and intense dilatation of the intercellular spaces. It was concluded that 20% ethanol provokes marked ultrastructural lesions in the hard palatine mucosa.

Morphology of the vas deferens in an ethanol-drinking strain of rats (UChA and UChB).

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of vas deferens, which are the subject of this analysis in UCh ethanol-drinking rats. The proximal and distal segments of the vas deferens of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium of the vas deferens and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the epithelium of the vas deferens and hypothalamus-pituitary axis of UCh rats.

Ultrastructure of the urethra of the Mongolian gerbil.

OBJECTIVE: The urethra is the main port of entry of sexually transmitted pathogens. However, papers on the morphology of the urethra are scarce. The Mongolian gerbil is a rodent native of the Mongolia and China and has been utilized as a laboratory animal since the 1960s. This work describes the ultrastructure of the urethra of the Mongolian gerbil to provide data for future experimental studies. METHODS: The urethra of ten adult male gerbils was studied by transmission electron microscopy. RESULTS: The epithelium of the pelvic urethra possesses two cell types: I and II, without the formation of cellular layers, while the penile urethra possesses cellular layers: basal, intermediate and superficial. The urethra presents neurosecretory cells belonging to the amine precursor uptake and decarboxylation system. CONCLUSIONS: The urethral epithelium of the gerbil is a neurosecretory epithelium, part of the amine precursor uptake and decarboxylation system.

Structure of the pelvic and penile urethra--relationship with the ducts of the sex accessory glands of the Mongolian gerbil (Meriones unguiculatus).

The urethra is the main place of entry for sexually transmitted pathogens. However, there is little literature on the morphology of the urogenital system, principally the urethra and ducts of the sex accessory glands. The Mongolian gerbil is an insectivorous, herbivorous and monogamous rodent with nocturnal habits; it has been used successfully as a laboratory animal since the 1960s. Therefore, the objective of the present paper was to describe the structure and ultrastructure of the urethra and its relations to the ducts of the accessory sex glands of the Mongolian gerbil (Meriones unguiculatus), contributing to the understanding of the reproductive biology of the rodent and aiming to provide data for future experimental studies. Conventional techniques of light and scanning electron microscopy were utilized. The urethra and ducts of the accessory sex glands are similar to those of the albino rat and the mouse. However, there is variation in drainage type among accessory sex glands for the inner urethra. The ducts of the seminal vesicle, the ductus deferens, drain their contents independently into the ampullary duct that opens in the urethra. The ducts of the prostate, coagulating and bulbourethral glands drain their contents independently into the urethra.

Kinetics of spermatogenesis in the Mongolian gerbil (Meriones unguiculatus).

The Mongolian gerbil (Meriones unguiculatus) is a small rodent native to the arid regions of Mongolia and Northeastern China. The present study provides descriptions of both the cellular associations of the seminiferous-epithelium cycle and relative frequencies of stages in the gerbil. Based on the development of the acrosomic system and the nuclear morphology changes using the PAS-H staining technique, the transformation of spermatids into spermatozoa was divided into 15 steps. The first 12 steps were used to identify 12 stages or cellular associations and the other three steps were spread among the first six stages of the cycle of the seminiferous epithelium. The relative frequencies found for stages I through XII were: 13.15; 8.06; 8.98; 6.48; 5.37; 6.71; 7.36; 7.45; 7.27; 5.83; 11.53 and 11.81, respectively. Stage I had the highest frequency while stage V proved the lowest frequency among the XII stages. The pattern of spermatogenesis is similar to those of rodents used as laboratory animals. The present description is the first for this rodent and provides the foundation for a variety of future studies of the testis in this animal.

Morphological effects on the hard palatine mucosa of Calomys callosus submitted to experimental chronic alcoholism.

The morphological effects of ethanol ingestion on the hard palatine mucosa of adult male Calomys callosus were observed. Twenty rodents were divided into two experimental groups: the control group received solid diet, Purina rat chow, and tap water ad libitum; the alcoholic group received the same solid diet and ethanol P.A. diluted 20% in water (v/v). After 270 days of treatment, all animals were sacrificed and the hard palatine mucosa were prepared for TEM and SEM methods. The epithelial cells of the alcoholic group showed some alterations like cytoplasmatic lipid droplets, pycnotic nucleus and increased mitochondrial size. The lamina propria also presented intense lipid droplets accumulation. The morphological changes suggested that chronic ethanol consumption was able to modify the integrity of the mucosa.

Ultrastructural study of acrosome formation in mongolian gerbil (Meriones unguiculatus).

Spermatogenesis is a complex and very well organized process lasting from 30 to 75 days in mammals. The spermatogenic process has been described mainly in laboratory mammals, such as the rat, while correspondent studies in wild animals are scarce. The gerbil (Meriones unguiculatus) is a small rodent native of the arid regions of Mongolia and China. Few reports are available on reproduction in the male Mongolian gerbil. The present study provides the first description of the ultrastructural alterations in spermatid cytoplasm and nucleus, with particular reference to acrosome formation in gerbils. The testes were processed by conventional transmission electron microscopy technique. Based on the development of the acrosomal system and changes in nuclear morphology, the transformation of spermatids in spermatozoon was divided into 15 steps. There were four phases in the spermiogenesis process in the gerbil: Golgi, cap, acrosomal and maturation phases. This provides the foundation for a variety of future studies of the spermiogenesis of this animal.