Screening for inhibitors which regulate the sperm serin proteases

CLEMENTI MARISA ALEJANDRA; BOARELLI PAOLA VANINA; MONCLUS MARÍA DE LOS ÁNGELES; FORNÉS MIGUEL WALTER; CESARI ANDREÍNA

Mendoza, 5 al 7 de Diciembre de 2008.

Congreso; XXVI Reunión Científica Anual de la Sociedad de Biología de Cuyo y I Reunión Científica Anual de la Dirección de Investigación, Ciencia y Técnica dependiente del Ministerio de Salud; 2008

Sociedad de Biología de Cuyo

The regulation of serin protease activity in spermatozoa was attributed to inhibitors found in seminal plasma, synthesized in accessory glands that play an important role on sperm capacitation. In the present work, we sought for serine protease inhibitors in bovine seminal plasma (BSP) and murine accessory glands. We obtained protein extracts from BSP and accessory glands from CF-1 mice. Small molecules were selected by filtration membranes (30kDa). The inhibitors were detected by reverse zymography and quantified by their ability to reduce trypsin activity over azocasein, considering as 100% the inhibitory activity of SBTI. BSP showed the highest inhibitory activity (85%). Seminal vesicle (SV) and prostate (P) had the highest inhibitory activity (98-96% respectively), whereas epididymis (E) and testis (T) had a significantly lower activity compared to the control (SBTI): 14,5  and 7 % (p<0,05), respectively. The reverse zymography developed 1 protein below 30 kDa in each sample analysed and curiously, 1-2 bands between 26-66 kDa in most of them, except the SV. In conclusion, we detected and preliminary characterized serin protease inhibitors in PSB and accessory glands that potentially regulate sperm serine proteases. Mass spectrometry identification of these proteins will be the next step