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Molecular Human Reproduction Vol. 2, NUMBER 12 pp. 929-935, 1996
© European Society of Human Reproduction and Embryology 1996

research-article

lmmunoelectron microscopical distribution of histones H2B and H3 and protamines during human spermiogenesis

Y. Prigent1, S. Muller2 and J.P. Dadoune1,3

1Groupe d'Etude de la Formation et de la Maturation du Gamète Mâle, Laboratoire d'Histologie JE MENESR 349, UFR Biomé;dicale, 45 Rue des Saints-Péres, 75270 Paris Cedex 06 2Institut de Biologie Moléculaire et Cellulaire du CNRS UPR 9021, 67084 Strasbourg, France

To whom correspondence should be addressed at: 3To whom correspondence should be addressed

The fine structural distribution of histones H2B and H3, and protamines were localized by means of specific antibodies and ultrastructural immunocytochemistry in nuclei of human spermatids and spermatozoa. The antibodies were used to detect the nuclear basic proteins on sections of testis and ejaculated spermatozoa by immunoelectron microscopy. A quantitative analysis of labelling density was performed on micrographs using an interactive image analysis system. The labelling density of somatic-type histones H2B and H3 and of their testis-specific variants was constant in the nuclei of young spermatids with round nuclei (stages 1–2), and then increased in intermediate spermatids (stages 3–4). Histone H3 labelling decreased at the end of the elongation phase (stage 5) while histone H2B labelling decreased in mature spermatids (stage 6) only. Spermatozoa were found to be weakly labelled by the anti-histone antibodies. The first signs of labelling of protamines and basic intermediate proteins appeared in spermatid nuclei at stage 4, increased further in stage 6 spermatids and persisted in all sperm nuclei. The present work shows that histone-to-protamine replacement occurs at the beginning of the spermatid maturation phase in human. However, histones are partially retained in mature spermatids and sperm nuclei.

histones/immunocytochemistry/nuclear proteins/protamines/spermatozoa


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