ULTRASTRUCTURAL STUDIES ON DEVELOPING FOLLICLES OF THE SPOTTED RAY TORPEDO MARMORATA

Light and ultrastructural investigations on sub-adult and adult sexually mature females, demonstrates that in Torpedo marmorata folliculogenesis starts in the early embryo and that the two ovaries in the adult contain developing follicles of various sizes and morphology. Initially, the follicle is constituted by a small oocyte, surrounded by a single layer of squamous follicle cells. The organization is completed by a basal lamina and, more externally, by a theca, that at this stage is composed by a network of collagen fibers. As the oocyte growth goes on, during previtellogenesis and vitellogenesis, the organization of the basal lamina and of the oocyte nucleus does not change significantly. The basal lamina, infact, remains acellular and constituted by fibrils intermingled in an amorphous matrix; the nucleus always shows an extended network of chromatin due to the lampbrush chromosomes, and one or two large nucleoli. By contrast, the granulosa (or follicular epithelium), the ooplasm, and the theca cells significantly change. The granulosa shows the most relevant modifications becoming multi-layered and polymorphic for the progressive appearance of intermediate and pyriform-like cells, located respectively next to the vitelline envelope, or spanning the whole granulosa. The appearance of intermediate cells follows that of intercellular bridges between small follicle cells and the oocyte so that one can postulate that, as in other vertebrates, small cells differentiate into intermediate, and then pyriform-like cells, once they have fused their plasma membrane with that of the oocyte. Regarding the ooplasm, one can observe as in previtellogenic follicles, it is characterized by the presence of intermediate vacuoles containing glycogen, while in vitellogenic follicles by an increasing number of yolk globules. The theca also undergoes significant changes: initially, it is constituted by a network of collagen fibers, but later, an outermost theca esterna containing cuboidal cells and an interna, with flattened cells, can be recognized. The role of the different constituents of the ovarian follicle in the oocyte growth is discusse

Surface glycoproteins bearing alpha-GalNAc terminated chains accompany pyriform cell differentiation in lizards

The present investigation demonstrates that in squamate reptiles, as already reported for Podarcis sicula (Andreuccetti et al., 2001), the differentiation of pyriform cells from small, stem follicle cells is characterized by the progressive appearance on the cell surface of glycoproteins bearing alpha-GalNAc terminated O-linked side chains. Using a lectin panel (WGA, GSI-A4, GSI-B4, PSA UEA-I, PNA, Con-A, DBA, LCA, BPA, SBA), we demonstrated that, during previtellogenesis, the pattern of distribution of DBA binding sites over the follicular epithelium dramatically changes. In fact, binding sites first appear in follicular epithelium at the time that small cells begin to differentiate; in such follicles, labeling is evident on the cell surfaces of small and intermediate cells. Later on, as the differentiation progresses, the binding sites also become evident on the cell surface of pyriform cells. Once differentiated, the pattern of the distribution of DBA binding sites over the follicular epithelium does not change. By contrast, during the phase of intermediate and pyriform cell regression, DBA binding sites gradually decrease, so that the monolayered follicular epithelium of vitellogenic follicles, constituted only by small cells, shows no binding sites for DBA. It is noteworthy that binding sites for DBA are present on small cells located in contact with the oocyte membrane, but not on those located under the basal lamina or among pyriform cells, and therefore not engaged in the differentiation into pyriform cells. This finding demonstrates that, in squamates, the pattern of distribution of alpha-N-GalNAc containing glycoproteins significantly changes during previtellogenesis, and that these modifications are probably related to the differentiation of small stem cells into highly specialized pyriforms

The analysis of endurance, speed and explosive strength in young football players

Introduction Endurance, speed and explosive strength are determinant factors of sporting performance for football players (Hoff & Helgerud, 2004). An evaluation of these abilities is fundamental in young subjects, who have still not completed their maturation, to plan an individualised and effective training program (Fernandez-Gonzalo et al., 2010). The aim of this study was to examine the level of variability in endurance, speed and explosive strength of lower limbs among young football players. Methods Sixteen male players with 5 years of experience playing football and participating in the “Giovanissimi Regionali” amateur championship (14.00 years old ± 7 months; 57.5 ± 6.66 kg weight, 167.25 ± 6.97 cm height, 13.74 ± 4.40 % fat mass; 86.32 ± 4.46 % lean mass; 63.06 ± 4.69 % muscle mass) were tested. Endurance and speed were evaluated with the Cooper test and 30 m sprint test, respectively. Sargent jump and standing broad jump test were used to measure the explosive strength of lower limbs. The variability percentage of anthropometric features and motor abilities was analyzed calculating the coefficient of variation (CV) within the group. Values were considered significant at p<0.05. Results The players showed a higher variability in weight than stature (CV: 11.59% vs. 4.17%). In particular, weight variation was mainly due to a higher difference in fat mass than muscle mass percentage (CV: 32.03% vs. 7.44%). The explosive strength of lower limbs measured during vertical jump (38.88 ± 4.62 cm) was the ability more changeable (CV: 11.88% vs. 8.67%, 8.57% and 4.83%) compared to the same measured by standing broad jump (194.69 ± 16.88 cm), endurance (2780.00 ± 238.36 m) and speed (4.54 ± 0.22 s). Conclusion The increased variability in the elevation of vertical jump might be associated with a higher variation in fat mass and coordinative abilities among young football players. On the other hand, the lower variability in muscular mass might explain the scarce difference in speed. In conclusion, the explosive strength of lower limbs appears to be the ability that depends mostly on age. This data is preliminary because this study is still in progress and extending on a larger scale. References Hoff, J. & Helgerud, J. (2004). Endurance and Strength Training for Soccer Players. Sports Med, 34, 165-180. Fernandez-Gonzalo, R., Souza-Teixeira, F., Bresciani, G., García-López, D. et al. (2010). Comparison of technical and physiological characteristics of prepubescent soccer players of different ages. J Strength Cond Res, 24,1790-8

Fine structure of Leydig and Sertoli cells in the testis of immature and mature spotted ray Torpedo marmorata

An ultrastructural investigation revealed the presence of true Leydig cells in the testis of sexually mature specimens of Torpedo marmorata. They showed the typical organization of steroid-hormone-producing cells, which, however, changed as spermatocysts approached maturity. In fact, they appeared as active cells among spermatocysts engaged in spermatogenesis, while in regions where spermiation occurred, they progressively regressed resuming the fibroblastic organization typically present in the testis of immature specimens. Such observations strongly suggest that these cells might be engaged in steroidogenesis and actively control spermatogenesis. Sertoli cells, too, appeared to play a role in spermatogenesis control, since, like Leydig cells, they showed the typical aspect of steroidogenic cells. In addition, the presence of gap junctions between Sertoli cells suggests that their activity might be coordinated. After sperm release, most Sertoli cells were modified and, finally, degenerated, but few of them changed into round cells (cytoplasts) or round cell remnants, which continued their steroidogenic activity within the spermatocyst and the genital duct lumen. From the present observations, it can be reasonably concluded that, in T. marmorata, spermatogenesis depends on both Leydig and Sertoli cells, and, as postulated by Callard (1991), in cartilaginous fish, the function of the Leydig cells as producers of steroids might be more recent and subsequent to that of Sertoli cells. In this regard, it is noteworthy that, in immature males, when Leydig cells showed a fibroblastic organization, Sertoli cells already displayed the typical organization of a steroidogenic cel

AN ULTRASTRUCTURAL STUDY ON THE VITELLOGENESIS IN THE SPOTTED RAY TORPEDO MARMORATA

The present investigation strongly suggests that in Torpedo the oocyte growth is not only due to the uptake of exogenous molecules, but also by the oocyte itself and the granulosa cells. The oocyte, starting from the early previtellogenic follicles (see also Mol. Reprod. Dev. 61 (2002) 78), synthesizes large amounts of glycogen. Later, as the oocyte growth goes on, the cytoplasm of granulosa cells progressively bears numerous islets of glycogen, which are also evident inside the intercellular bridges and in the oocyte cortex, suggesting that they may flow from granulosa cells to the oocyte. The contribution of granulosa cells seems to become most relevant during the vitellogenesis. In vitellogenic follicles, both small, intermediate, and pyriform-like cells bear numerous vacuoles containing vitellogenin-like material, suggesting strongly that in Torpedo, differently from other vertebrate species, granulosa cells could be engaged in vitellogenesis. The present investigation does not allow us to know if such a material is due to a transcytosis process and/or is synthesized inside them. The organization of granulosa seems to exclude the possibility that it is transferred to granulosa via transcytosis. On the contrary, granulosa cells, especially in vitellogenic follicles, display the morphological organization of metabolically active cells, so they could be engaged in vitellogenin synthesis. This interpretation is consistent with the observation that granulosa cells are positively stained by OZI (osmium tetroxide-zinc iodide) and that the same positivity is evident on intercellular spaces, containing vitellogenin-like material, and on nascent yolk globule