1,721,011 research outputs found
Attivazione del recettore c-kit ed incremento dell’attività protein chinasica mediata dal D-Asp nel testicolo di Podarcis sicula
No full textD-Asp: A new player in reproductive endocrinology of the amphibian Rana esculenta
No full textWe investigated the involvement of D-Aspartic acid (D-Asp) on ovarian and testicular morphology of the green frog, Ram esculenta, and its effect on the testosterone production. The study has been performed throughout the reproductive cycle. In both ovary and testis a substantial amount of D-Asp is endogenously present and its concentration varies as function of reproduction. In the frog, D-Asp content is differently correlated with gonadal and plasmatic levels of testosterone, depending on the sex. In fact, the amount of the D-Asp is inversely linked with that of the testosterone in the ovary, while this correlation directly matched in the testis. In vivo short-term experiments, consisting of a single intra-peritoneal injection of D-Asp (2.0 mu mol/g body weight), demonstrated that the enantiomer is significantly accumulated by both the ovary and testis, reaching after 3 h the highest uptake and thereafter decreasing to baseline values within 24 h. Furthermore, D-Asp influences the synthesis and/or the release of testosterone, causing a decrease of its level in the female, and an increase in the male, respectively. In vivo long-term experiments, D-Asp, chronically administered to the frogs of both sexes, enhances the maturation of both gonads, determining in the oocytes an higher accumulation of carbohydrate yolk plates in the ooplasm, and stimulating the spermatogenesis in the testis. Taken altogether, our results show that D-Asp operates differently in female and male frog gonads, indicating that it has different targets in the reproductive machinery depending on the sex. (C) 2011 Elsevier B.V. All rights reserved
Analisi comparativa del ruolo dell’acido D-aspartico sul controllo della steroidogenesi nei vertebrati
No full textLocalization of c-kit and stem cell factor (SCF) in ovarian follicular epithelium of a lizard, Podarcis s. sicula
No full textWe performed an immunohistochemical study to determine the immunolocalization of c-kit and stem cell factor (SCF) in ovarian follicles during the reproductive cycle of the lizard, Podarcis s. sicula. Follicles were serially cut and used for histological and histochemical characterization and also for immunohistochemical detection of both c-kit and SCF. C-kit and SCF were localized in the follicles with a differing pattern with regard to the stage of sexual cycle or the cell type forming the follicular epithelium (granulosa). In pre-reproductive follicles, where the granulosa consists of three main different cytotypes, the c-kit receptor was prevalently localized on the plasmalemma of small cells, although some pyriform and intermediate cells also appeared positive. C-kit was also localized in the theca. In pre-reproductive follicles, SCF was markedly observed in the cytoplasm of some pyriform cells. Small cells and theca also stained moderately positive, whereas the intermediate cells were mostly negative. In reproductive follicles, where granulosa cells are morphologically rearranged, c-kit was observed in small cells and in some thecal elements, while SCF was weakly immunostained. At the site of follicular layer invaginations evident c-kit/SCF immunostaining was observed in the granulosa epithelium and in the theca. These observations suggest that the expression of c-kit and SCF changes as a function of follicular development and may reflect the involvement of this system in the maturation of the oocyte. © 2010 Elsevier GmbH
The maturation of oocyte follicular epithelium of Podarcis s. sicula is promoted by D-aspartic acid
No full textWe investigated whether the maturation of oocyte follicular epithelium of lizard is affected by D-aspartic acid (D-Asp). Our results demonstrated that D-Asp is endogenously present in the oocytes, and its distribution varies during the reproductive cycle and following intraperitoneal administration. At previtellogenesis, it is observed in the cytoplasm and nucleus of pyriform cells, in intermediate cells, in some small cells of the granulosa, in the ooplasm, and in some thecal elements. At vitellogenesis, D-Asp is localized in the proximity of the zona pellucida, in the theca, and in the ooplasm. Injected D-Asp is mainly captured by pyriform cells and ooplasm of previtellogenic oocytes, but a moderate accumulation is evident in the cytoplasm of some small granulosa cells and in the theca. D-Asp also increases the ovarian and plasmatic levels of 17β-estradiol and decreases those of testosterone. As a direct and/or indirect consequence of D-Asp, previtellogenic oocytes grow up and mature, resulting in a higher accumulation of carbohydrates in the granulosa, zona pellucida, and ooplasm, but also a reduction in the thickness of the granulosa layer and an increase of the theca stratum. Taken together, our results show that D-Asp may be related to the synchrony of reproduction, either enhancing the growth and maturation of follicular epithelium or influencing its endocrine functions. © 2010 Raucci and Di Fiore
The c-kit receptor protein in the testis of green frog Rana esculenta: Seasonal changes in relationship to testosterone titres and spermatological proliferation
No full textThe green frog Rana esculenta is a seasonal breeder. The cyclic changes between almost arrested and highly activated spermatogenesis offer an ideal model to study basic mechanisms of spermatogenesis. In this study, we demonstrated, to our knowledge for the first time, c-kit receptor positive cells in the testis of this amphibian. The presence of c-kit receptor protein was confirmed by western blotting (Wb) analyses carried out in the testis during all the three main phases of the sexual cycle. The antibody recognized a band of about 150 kDa that was correlated with the positive staining in the germinal epithelium. The immunolabelling for c-kit receptor, evaluated by immunohistochemistry (IHC), was localized in I and II spermatogonia (SPG), in I and II spermatocytes, in both elongating spermatids and spermatozoa and in the Leydig cells. Furthermore, c-kit expression showed a seasonal pattern connected with both testicular and plasma profiles of testosterone during the reproductive cycle. The highest expression of c-kit receptor occurred during the reproductive period, when the testis exhibited the maximum concentration of testosterone. In this period, the mitotic activity of germ cell, assessed by both Wb and IHC analyses for proliferating cell nuclear antigen (PCNA), was intensive. indeed, during the post-reproductive period, testosterone titres were the lowest and the expression of both PCNA and c-kit receptor protein in the testis, although present, is minor when compared with the reproductive phase. This evidence suggests that cell division can continue sufficiently to accumulate SPG for the next spring, when new germinal cells undergo multiplication. Finally, during the pre-reproductive period, testosterone levels begin to increase and mitotic activity of germinal epithelium is comparably enhanced. These events seem to precede the period of maximum stimulated spermatogonial proliferation, i.e. the reproductive period. These results suggest that the c-kit receptor may play a role in germ cell proliferation and provide a basis for future detailed investigation of regulatory factors of the proliferation of SPG. © 2007 Society for Reproduction and Fertility
The reproductive activity in the testis of Podarcis s. sicula involves d-aspartic acid: A study on c-kit receptor protein, tyrosine kinase activity and PCNA protein during annual sexual cycle
No full textThe current study provides substantial evidence that the pattern of synthesis of d-aspartic acid (d-Asp) in the testes of lizard Podarcis s. sicula throughout the reproductive cycle is in parallel with seasonal variations of testosterone, c-kit receptor protein, tyrosine kinase activity, and proliferating cell nuclear antigen (PCNA) protein. Although the trend is the same in all phases of the sexual cycle, the peaks of these three molecules are detectable only during the reproductive period. Using Western blot technique, we demonstrated that both polyclonal c-kit and PCNA antibodies specifically recognized bands with molecular mass of ∼150 and ∼36 kDa, respectively. By immunocytochemical methods, d-Asp immunopositivity appeared spread in the germinal epithelium as well as in the interstitial compartment of the testes. We also found specific c-kit labeling in I and II spermatogonia (SPG), in I and II spermatocytes (SPC), in the elongated spermatides, in spermatozoa, in Sertoli and Leydig cells. Like c-kit, PCNA positivity was located in the germinal epithelium pattern. Furthermore, we investigated the relationship between testosterone, c-kit receptor, tyrosine kinases activity and PCNA following treatment with d-Asp. In vivo experiments, entailing a single injection of d-Asp (2.0 μmol/g body weight), demonstrated that this amino acid significantly accumulated in the testes. After 3 h, its uptake was accompanied by an increase in testosterone levels and in the expression and intensity of immunostaining of c-kit receptor protein. Furthermore, at 6 h, exogenous d-Asp affected the phosphorylation of tyrosine kinases, whose activation was positively correlated with the temporal uptake of both d-Asp and testosterone detected in the testes. Thereafter, between 6 and 15 h, the expression of PCNA was induced and an increase in its immunolabeling intensity was observed. Taken all together, these results provide new insights into the testicular activity during the reproductive cycle of Podarcis s. sicula, suggesting that a sequential cascade of a functional relationship between testosterone levels, c-kit receptor protein, tyrosine kinase activity and PCNA could be partly mediated by d-aspartic acid. © 2009 Elsevier Inc. All rights reserved
Steroidogenic gene expression following D-aspartate treatment in frog testis
No full textPrevious studies have provided evidence that D-Asp plays a role in steroid-mediated reproductive biology in amphibians, reptiles, birds and mammals. To examine the molecular involvement of D-Asp on steroidogenic pathway regulation, we analysed the expression of StAR, P450 aromatase and 5 alpha Red2 mRNAs in Pelophylax esculentus testis, either in relation to the reproductive cycle or D-Asp treatment. Basal StAR mRNA levels, as well as D-Asp and testosterone concentrations, were higher in reproductive than in post-reproductive frogs. D-Asp treatment increased StAR mRNA expression and immunolocalisation in both the reproductive and post-reproductive periods. In control testis, aromatase mRNA levels were higher in the post-reproductive period, but following D-Asp administration, they increased only in the reproductive period. The level of 5 alpha tRed2 mRNA was higher in reproductive frogs than in post-reproductive frogs, and it increased after D-Asp treatment only in the post-reproductive phase. Our results suggest that, in P. esculentus testis, D-Asp increases StAR mRNA in both periods, and P450 aromatase and 5 alpha Red2 mRNAs at different points during the reproductive cycle. (C) 2011 Elsevier Inc. All rights reserved
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