Reptilian epididymis is considered as an important excurrent duct system required for the sperm maturation. Reptilian epididymis synthesizes and secretes proteins which vary in different regions of the epididymis. Hence, to investigate the effect of the secretions of different regions of the epididymis on spermatozoa motility, an in vitro study was undertaken to observe the changes in the patterns of motility of the testicular spermatozoa incubated with luminal contents of different regions of the epididymis in the lizard Eutropis carinata for the first time. The non motile testicular spermatozoa from the testis exhibited different patterns of motility, when incubated with the luminal contents of different regions of the epididymis. The spermatozoa from the testis, different regions of the epididymis exhibited 8 different patterns of motility (a-h). Testicular spermatozoa incubated with the anterior and middle epididymal luminal contents showed the motility patterns almost similar to that of the spermatozoa of the anterior and middle epididymis respectively. In contrast to the spermatozoa of the posterior epididymis, none of the testicular spermatozoa showed any movement when incubated with the posterior epididymal luminal contents. This study throws light on the importance of each region of epididymis in the physiological maturation of spermatozoa.
Effect of chlorpromazine (dopamine receptor antagonist) on pre-pubertal ovarian follicular development and onset of puberty were studied. Fifteen days old female rats were administered chlorpromazine (2.5 mg /kg body weight) daily for 21 days and appropriate controls were maintained. The onset of puberty in immature female rats was delayed following chlorpromazine treatment. There were significant increases in the body weight, ovary weight and diameter of the ovary of controls and treated group over the initial controls. Ovary of the initial controls consisted of primordial (type 2), primary (type 3a, 3b), pre-antral (type 4, 5a, 5b) and antral (type 6) follicles whereas, antral (type 7) and pre-ovulatory follicles (type 8) were not developed. Controls and treated group consisted of all types of follicles i.e. primordial to pre-ovulatory follicles. Primordial follicles were reduced in number significantly in the ovary of the controls and treated group when compared with the initial controls whereas there was no significant variation among the controls and the treated group. The mean number of primary, pre-antral and antral (type 6) follicles in the control and treated group increased significantly over the initial controls. However, there was a significant reduction in the mean number of these follicles in the treated group when compared to controls. The mean number of type 7 (antral) and type 8 (pre-ovulatory) follicles were reduced in the treated group when compared with controls. The number of atretic follicles of the primary, pre-antral and antral (type 6) follicles significantly increased in controls and treated group over the initial controls. When compared to controls the mean number of atretic follicles belonging to primary, pre-antral, antral (type 6 and 7) and pre-ovulatory category were significantly higher in treated group and the number of corpora lutea was significantly lower. The results indicate that chlorpromazine effect results in loss of follicles by atresia and delay the onset of puberty in immature female rats.
The epididymis of the male reproductive system is known to be involved in sperm maturation via the production of polypeptides, glycoproteins, surface proteins, enzymes and other factors. During the annual reproductive cycle, the epididymis of the lizard Eutropis carinata undergoes dramatic changes, both morphologically and biochemically, that occur in a well-organized sequence. The present study reveals the sequential changes that occur in the production and concentration of proteins in the epididymal luminal fluid throughout the annual reproductive cycle. A one-dimensional electrophoretic profile of the epididymal luminal proteins revealed a total of 18 bands in the regenerative phase, 22 bands during breeding and 17 bands in the post-breeding as well as regressed phases of the reproductive cycle. By two-dimensional electrophoresis, the protein complexes that are unique to the breeding phase were further resolved based on their pI and the molecular weight of each protein of the protein complex was determined. This is the first study to observe that proteins that are present during the reproductively inactive phase disappear during the reproductively active phase. The Periodic Acid Schiff (PAS) test for protein profiles revealed the presence of proteins with a carbohydrate moiety. Certain enzymes, such as acid phosphatase, alkaline phosphatase, and α-glucosidase, are highly sensitive to seasonal changes and their activity parallels the production of the epididymal proteins. This study provides evidence for androgen-dependent cyclical changes in the pattern of protein profiles and enzyme activity of the epididymal lumen in the lizard E. carinata.
Beta-hexosaminidase (Hex) is the major lysosomal enzyme associated with the event of fertilization. In this study, we have analyzed the distribution of Hex in the testis and the epididymis of the lizard, Eutropis carinata by a polyclonal antibody of β-hexosaminidase isoform (Hex A). Presence of Hex in the epididymis was performed by Western blotting. The result reveals that Hex A is present in the epididymal epithelium, lumen as well as spermatozoa. The anatomical distribution of Hex was studied by immunohistochemical localization. The study reveals that Hex is intensely stained in the epithelium of anterior and middle regions of the epididymis, whereas, posterior epididymal epithelium shows moderate staining. In addition, seminiferous epithelium of the testis shows staining for Hex. But lumen of the testis did not show any reaction for Hex. Further, immunohistochemical localization of Hex on the spermatozoa from the testis and different regions of the epididymis revealed that the Hex from the testis did not show any staining; the epididymal epithelium is moderately localized in the spermatozoa of the anterior region and gradually increases in the intensity in the spermatozoa of the posterior region of the epididymis. This indicates that the Hex is released from the epididymal epithelium and binds to the spermatozoa, and in the lumen, it gradually increases from anterior to the posterior region of the epididymis. The result also suggests that Hex A bound to the epididymal spermatozoa originates from the epididymis and not from the testis. The regional difference in the expression of Hex in the epididymis of the lizard, E. carinata, indicates the possible site of secretion of this enzyme.
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