The epididymis of the prepubertal swamp buffalo (Bubalus bubalis): histochemistry of phosphatases.

The distribution of phosphatases was examined in the epididymis of swamp buffaloes aged 1.5-2 years, where spermatogenic activity in the testis had reached the early primary spermatocyte stage. A granular distribution of acid phosphatase was present in the luminal region of the epithelium of the efferent ductules. In the ductus epididymidis, four zones were identified and all zones showed varying degrees of acid-phosphatase activity in the epithelium, with the most pronounced activity in the apical region and stereocilia of zone II. Alkaline-phosphatase activity occurred along the basal region of the epithelium in zones I, III and IV of the ductus and in the stereocilia of zones I and II. An intense apical reaction was seen in zone II. The efferent ducts were free of this enzyme. Thus, zone II is considered the most active region, having the function of absorption and possibly steroid metabolism.

Influence of media and thermal shock on sperm capacitation.

To elucidate the influence of medium and thermal shock in inducing sperm capacitation, eight ejaculates were processed as follows. Each ejaculate was divided into three aliquots. To two of the aliquots, an equal volume of saline was added, while TEST-yolk was added to the remaining aliquot. All three samples were cooled slowly to 5 degrees C and incubated for 18-20 h. After aspirating the supernatant, 6 ml of Tyrode solution containing 0.06 g% egg yolk, warmed to 37 degrees C was added without disturbing the sperm pellet to the aliquot containing TEST-yolk and to one of the aliquots mixed with saline. To the remaining saline-processed sperm pellet, 6 ml TEST-yolk (37 degrees C) was added. All samples were washed twice (400 x g for 5 min) and processed for the sperm penetration assay (SPA). Samples, either pre-incubated in TEST-yolk (mean +/- SEM; 65.9 +/- 11.7%) or warmed (thermal shock) with TEST-yolk (62.6 +/-12.1%) yielded significantly (P < 0.05) higher SPA outcome than the samples processed and warmed with saline (17.8 +/- 12.0%). The penetration indices were not statistically different from each other. It therefore appears that the medium influences the sperm capacitation process to a greater extent and may augment the influence of thermal shock.

Pregnancy terminating effect of Jatropha curcas in rats.

The fertility regulatory effect of the fruit of Jatropha curcas was investigated by oral administration of different extracts to pregnant rats for varying periods of time. Foetal resorption was observed with methanol, petroleum ether and dichloromethane extracts indicating the abortifacient properties of the fruit. The results also suggest that the interruption of pregnancy occurred at an early stage after implantation. This effect could be accomplished even when the extracts were given from the 6th to the 8th day of pregnancy. Loss of body weight during the dosing period, ranging from slight to severe was seen in the treated animals. Marked toxicity was observed with some extracts when given over a comparatively long period of about 10 days.

TEST-yolk media and sperm quality.

Media prepared with egg yolk and two buffers, TES and Tris, called TEST-yolk have been shown to have beneficial effects on the survivability, fertilizing capacity and storage potential of human spermatozoa. Egg yolk lipoproteins are the critical compounds for the beneficial effects, with a synergistic effect due to the TES-Tris buffer component. TEST-yolk media have been used in the sperm penetration assay, the hemizona assay, sperm preparation for clinical in-vitro fertilization, artificial insemination with homologous spermatozoa, cryopreservation, sperm samples with a positive antisperm antibody test, and preparation for techniques requiring capacitated spermatozoa. Few harmful consequences of TEST-yolk have been reported, although controlled trials are required to evaluate therapeutic effects in the treatment of male infertility.

Ultrastructural localisation of alkaline phosphatase in the intertubular tissue of the testis in the domestic fowl.

Alkaline phosphatase activity in the intertubular tissue of the testes of the domestic fowl was examined using an ultracytochemical technique based on the lead capture method. In the interstitial tissue, the Leydig cells, transitional cells and the fibroblasts displayed enzyme activity on their cell membranes. Vacuoles located in the transitional cells were lined by reaction products of enzyme activity, whereas the vacuoles representing extracted lipid droplets and present mainly in the Leydig cells were free of enzyme activity. In the peritubular tissue the cell processes of fibroblasts showed enzyme activity on the cell membranes and in pinocytotic vesicles. Cell processes lying adjacent to blood vessels showed pronounced activity. In the blood vessel itself some activity was present in the basement membrane and the endothelium. The surface of the red blood cell showed moderate activity. The possible role of alkaline phosphatase in the transfer of hormone from the Leydig cells to the seminiferous tubules and from the seminiferous tubules to the interstitium is discussed. The myoid cells and their processes were devoid of enzyme activity.

Ultrastructural localisation of alkaline phosphatase in the testis of domestic fowl.

The ultrastructural distribution of alkaline phosphatase has been investigated in the testis of the domestic fowl. Enzyme activity was observed in the germinal epithelium and in the peritubular tissue. In both situations the activity was located in the plasma membranes. The activity in the germinal epithelium was marked basally in the germinal cells and in the Sertoli cells. There was a gradual reduction and an ultimate disappearance of enzyme activity towards the luminal region. The presence of enzyme in the germinal epithelium is in contrast to results of similar studies in some mammals. The peritubular localisation is comparable to that of mammals and confirms light microscope observations reported for the fowl testis.

Ultrastructural localization of phosphatases in the testes of the domestic fowl: acid phosphatase and thiamine pyrophosphatase.

ACPase and TPPase activity has been examined in the germinal epithelium of the testes in the domestic fowl. ACPase activity in spermatogonia and spermatocytes was confined to the Golgi complex. In spermatids ACPase activity was seen in the endoplasmic reticulum and nuclear envelope in the phase I and especially in the phase II (the elongating phase). This activity gradually decreased during the next phase III (the elongating phase). This activity gradually decreased during the next phase III, and had disappeared in the final phase IV. The membrane body showed ACPase reaction in the small peripheral vacuoles and cisternal structures surrounding large central vacuoles. ACPase was also present in vesicles surrounding the developing tail. Late spermatids showed an abundance of autophagic vacuoles which had a complex array of ACPase positive delimiting membranes. In Sertoli cells ACPase activity was predominant in the lysosomes. TPPase activity was seen in the cisternae of the Golgi complex in spermatogonia and spermatocytes. In spermatids activity was present in the endoplasmic reticulum during the phase II, but it is lost in later stages. The smaller vacuoles and cisternal structures in the membrane body also showed reaction products. According to the present results it is thought likely that the smaller vacuoles and cisternal structures of the membrane body are of endoplasmic reticulum origin. The autophagic vacuoles in spermatids and the lysosomes of Sertoli cells are considered responsible for the degradation of residual bodies cast off by spermatids.

On the structure of the vascular body in the domestic fowl.

The microscopic structure of the vascular body has been described. Three cell types have been identified. The arrangement of blood and lymph vessels has been demonstrated by means of injection techniques, and these vessels have been investigated at an ultrastructural level. The function of the vascular body is discussed.

Ultrastructural studies on the differentiation of spermatids in the domestic fowl.

Four phases of spermatid maturation are recognized on the basis of nuclear morphology. The formation of the acrosome, perforatorium and axial filament complex are described in relation to these phases. The functional significance of the microtubular systems in nuclear elongation and spermatid curvature are discussed. A membranous structure, hitherto unidentified, and referred to in this study as the 'membrane body' is compared with the chromatoid body. Morphological evidence suggests that this structure, together with the annulate lamellae, removes excessive nuclear material.