GIAMBELLUCA M.A.

GIANNESSI F

RUFFOLI R.

English

RUFFOLI, RICCARDO

GIAMBELLUCA, MARIA

GIANNESSI, FRANCESCO

Archivio della Ricerca - Università di Pisa

ULTRASTRUCTURAL STUDY OF THE NADPH-DIAPHORASEACTIVITY IN MICE AGEING TESTESRUFFQLI R.. GIAMBELLUCA M. A., GIANNESSI F.Dipartimento di Morfologia Umana e Biologia Applicata - Facolta di Medicina eChirurgia Universita degli Studi di PisaINTRODUCTION: A decline of testicular function occurs with ageing in mammals.Diminished sperm production and decreased testosterone secretion were in factshown in humans and in laboratory rodents (1). A number of morphological altera-tions were also described in the Leydig cells and in the seminiferous tubules. Themost frequent Leydig cell modifications consisted of poor development of the SERand of reduced both volume and number of the mitochondria. In addition, severalLeydig cells exhibited lipofuscin granules in their cytoplasm. Recently, nitric oxidesynthase (NOS), the enzyme responsible for the production of nitric oxide (NO), wasrecognized in the Leydig cells (2). Results so far achieved show NO to suppress thetestosterone secretion (3). Modifications of NO production were described in severalorgans of ageing mammals. Therefore, the puipose of this study was to examine byTEM the distribution of the NOS enzymatic activity in the Leydig cells of ageingmice.MATERIAL AND METHODS: Ten Swiss mice aged 12 months were anaestheti-sed, perfused by aldehydes and treated for NADPH-d histochemistry (4). In a pre-vious study we demonstrated that NADPH-d reaction overlaps with NOS in the Ley-dig cells of mouse (5). The specimens were post-fixed in osmium tetroxide andembedded in Epon 812, following the usual procedures.RESULTS AND DISCUSSION: In the ageing mice the majority of the Leydig cellswere smaller in size than those of young adults, and their shape was elongated. Thesecells contained a small number of lipid droplets and poor developed both SER andmitochondria. In addition the NADPH-diaphorase reaction was strongly increased inrespect to young animals. The reaction product was widely spreaded in the SER andseveral mitochondria were labeled. Leydig cells were also present filled with lipidvacuoles, showing a distribution of the enzymatic reaction like that observed inyoung animals. Some Leydig cells contained several lipofuscin granules. The presentstudy indicates that most of Leydig cells of ageing mice undergo increase of NOSenzymatic activity, and suggests that NO could be involved in the age-associateddecline of testosterone secretion.REFERENCES:1) ZIRKIN B. R. et al, Exp Gerontol 1997, 32: 529-537.2) DAVIDOFFM. S.etal, Cell Tissue Res 1997,287: 161-170.3) DEL PUNTA K. et al., Endocrinology 1996, 137: 5337-5343.4) WOLF G. et al., Neurosci Lett 1992, 147: 63-66.5) GIANNESSI F. et al., Histochem Cell Biol 1998, 109: 241-248.189

Ultrastructural study of the NADPH-diaphorase activity in mice ageing testes

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Ultrastructural study of the NADPH-diaphorase activity in mice ageing testes

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