Human Reproduction, Vol 13, 1559-1566, Copyright © 1998 by Oxford University Press
HJ van Roijen, MP Ooms, MC Spaargaren, WM Baarends, RF Weber, JA Grootegoed and JT Vreeburg
During mammalian spermatogenesis, the chromatin of the spermatogenic cells
is profoundly reorganized. Somatic histones are partly replaced by
testis-specific histones. These histones are then replaced by transition
proteins and finally by protamines. This series of nucleoprotein
rearrangements results in a highly condensed sperm cell nucleus. In
contrast to spermatozoa from other species, human spermatozoa still contain
a significant amount of histones, including testis-specific histone 2B
(TH2B). In the present study it is shown that an antibody targeting
tyrosine hydroxylase, which has been found previously to cross-react with
rat TH2B, also specifically immunoreacts with human TH2B on Western blots,
in immunohistochemistry of human testis tissue, and in immunocytochemistry
of decondensed human spermatozoa. In human testis tissue, TH2B
immunostaining first apparent in spermatogonia, shows marked variation,
especially at the pachytene spermatocyte stage, and then reaches an intense
signal in round spermatids. Shortly before spermatid elongation, a portion
of the spermatid nucleus, corresponding to the acrosomal region, loses its
immunoreactivity. During condensation of the spermatid nucleus, the
immunodetectability of TH2B disappears gradually, from the anterior region
of the nucleus onwards. At the final stages of spermiogenesis, the
immunostaining is completely absent. Immunocytochemical staining of
spermatozoa revealed no TH2B immunosignal, but immunostaining was observed
when spermatozoa obtained from semen were decondensed to make nuclear
proteins accessible to the antibody. There was, however, a striking
intercellular variability in the intensity of staining of spermatozoa
within an ejaculate. In a population of 35 men attending our Andrology
Clinic, we observed interindividual differences in total sperm TH2B
content, which showed a significant, although not very pronounced, negative
correlation with normal morphology (P = 0.05).
ARTICLES
Immunoexpression of testis-specific histone 2B in human spermatozoa and testis tissue
Department of Endocrinology and Reproduction, Faculty of Medicine and Health Sciences, Erasmus University Rotterdam, The Netherlands.
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