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P Chieffi, A Picascia, R Stanzione, and D Tramontano

Proto-oncogenes play an important role in the regulation of cellular growth and differentiation. C-Jun activity has been studied in the testis of a non mammalian vertebrate, the lizard Podarcis s. sicula, during two different periods: winter stasis and the breeding season. C-Jun protein was localized by immunocytochemistry in the cytoplasm of the spermatogonia (SPG) and stage I and II spermatocytes (SPC) during the winter stasis (from December until March), while the protein was present in the nuclei of the same cells during the active spermatogenic period (April/May). The different localization of c-Jun has been confirmed by Western blot and immunoprecipitation analysis. In addition, when Jun is present in the nuclear compartment, it is phosphorylated on Ser-63 and is complexed with Fos protein.These data suggest that the nuclear localization of the Jun protein in the SPG and stage I and II SPC, with strong phosphorylation on Ser-63 during the breeding period, could be the signal of increasing transcriptional activity in the lizard testis.

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P Chieffi, GL Colucci-D'Amato, S Staibano, R Franco, and D Tramontano

Several lines of evidence support a key role of estradiol-17beta (E(2)) in male fertility. We have used a non-mammalian vertebrate model, the frog Rana esculenta, to investigate the regulation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) activity in the testis during the annual sexual cycle and to study whether E(2 )exerts a role in spermatogenesis through the regulation of ERK1/2 activity. ERK1/2 proteins are present in the cytoplasm and nucleus of the primary and secondary spermatogonia (SPG), and in the nucleus of primary spermatocytes. The annual E(2) profile shows a progressive increase during active spermatogenesis with a peak in the month of June. In parallel, ERK1/2 are highly phosphorylated during the period of active spermatogenesis (from April to July) compared with the regressive period (September/October) and winter stasis (from November to March). E(2) treatment induces the proliferation of primary SPG, possibly via the activation of ERK1/2, and this effect is counteracted by the anti-estrogen ICI 182-780.

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P Chieffi, G Troncone, A Caleo, S Libertini, S Linardopoulos, D Tramontano, and G Portella

Aurora/Ipl1-related kinases are a conserved family of proteins that have multiple functions during mitotic progression. High levels of Aurora kinases are characteristic of rapidly dividing cells and tumours. Aurora B encodes a protein that associates with condensing chromatin, concentrates at centromeres, and then relocates onto the central spindle at anaphase. In this study the expression and the localisation of Aurora B throughout germinal epithelial progression in normal testis and its neoplastic counterpart were analysed. Immunocytochemistry and RT-PCR analysis of mouse germinal epithelium cells showed the presence of Aurora B in spermatogonia and occasionally in spermatocytes. Western blot analysis revealed the typical Aurora B isoform ( approximately 41 kDa) in the same cellular types. A similar distribution was observed in human testis by immunohistochemistry. Moreover, the distribution and the expression of Aurora B were investigated in neoplasms derived from germ cells. Surgical samples of seminomas were analysed, and a high percentage of Aurora B positive cells (51%) was detected; the expression of Aurora B was significantly related to the MIB-1 proliferation marker (R=0.816). The data presented here demonstrate that Aurora B expression occurs in spermatogonial division. Furthermore, our results indicate that the expression of Aurora B is a consistent feature of human seminomas.