Effect of age on seminiferous tubule protein secretion and the adverse effects of testicular toxicants in the rat

Abstract

This study has assessed the effect of age on protein secretion by seminiferous tubules (ST) isolated from rats and their response to Sertoli cell toxicants. ST were isolated from immature (aged 28 days), late pubertal (aged 45 days) and young adult (aged 70 days) rats and cultured in vitro for 24 h with 35S-methionine in the presence or absence of FSH (1 mg/ml), m-dinitrobenzene (m-DNB) or nitrobenzene (NB) (both at 10(-4)M). Incorporation of 35S-methionine into newly synthesized proteins in the culture medium (secreted proteins) was assessed and the pattern of protein secretion evaluated using two-dimensional sodium dodecylsulphate polyacrylamide gel electrophoresis (2-D SDS-PAGE). These data were compared with those obtained using cultures of immature rat Sertoli cells+germ cells (SC+GC). Addition of FSH, m-DNB or NB in vitro either had no effect (NB) or had a small stimulatory effect (FSH and m-DNB) on the incorporation of 35S-methionine into overall secreted proteins by ST isolated from immature rats. At the same doses, addition of either FSH, m-DNB or NB to SC+GC co-cultures resulted in increased incorporation of radiolabel into secreted proteins in all instances. In contrast, the same additions to ST isolated from adult rats resulted in a 20-34% decrease in the overall incorporation of 35S-methionine into secreted proteins. ST isolated from late pubertal rats showed a similar response to adult rats except that the decreases in incorporation induced by FSH, m-DNB and NB were smaller. Analysis by 2-D SDS-PAGE revealed considerable age-dependent differences in the proteins secreted by ST from immature and adult rats, of which 13 were identified as being of potential importance. Most of these proteins were prominent secretory products of ST from adult rats, but were minor or non-detectable products of cultures of ST or SC+GC from immature rats. Most of these proteins disappeared or decreased in abundance after culture of ST with m-DNB or NB. Two proteins showed the reverse pattern, being more prominent secretory products in immature than mature rats, and their secretion was unaffected or was increased by toxicant exposure. These results demonstrate that there are major age-dependent differences in the secretion of total and specific proteins by isolated ST and that these are probably related to changes in the germ cell complement with age.(ABSTRACT TRUNCATED AT 400 WORDS)