PROTEIN AND DNA CONTENTS IN SPERM FROM AN INFERTILE HUMAN MALE POSSESSING PROTAMINE DEFECTS THAT VARY OVER TIME

Sperm from 2 semen samples collected 6 months apart from an infertile male and 3 semen samples collected over an 18-month period from a fert ile human male volunteer have been analyzed for their protamine and DN A content. Hup1M and Hup2b antibodies were used to detect the presence of protamines and protamine precursors in western blots of nuclear pr oteins isolated from pools of sperm. Phosphorus and sulfur contents, w hich can be used to estimate the nuclear DNA and protamine contents of sperm from fertile males, were measured within individual sperm heads from each semen sample by particle induced x-ray emission (PIXE). The single-cell data reveal no significant differences in the phosphorus and sulfur contents of sperm heads in the three semen samples obtained from the fertile male. For the initial semen sample produced by the i nfertile male, Western blot data show a normal complement of protamine 1, small amounts of mature protamine 2, and reveal large amounts of a nti-protamine 2 reactive proteins with electrophoretic mobilities simi lar to protamine 2 precursors. Data from PIXE show elevated levels of sulfur within sperm heads compared with sperm from the fertile male. W estern blot data exhibit no evidence of protamines or protamine 2 prec ursors in the second semen sample produced by the infertile male. Data from PIXE suggest that these sperm are highly deficient in sulfur and protamines. These results show that the degree of maturation of sperm cells present in the semen of some infertile males can vary with time . (C) 1998 Wiley-Liss, Inc.