title: Sperm chromatin protamination: an endocrine perspective.
abstract: During spermiogenesis, the elongating rat spermatid chromatin undergoes a gradual process of condensation which is initiated in the round spermatids at "step 7" of cytodifferentation in stage VII and extending to elongated spermatids at "step 19" of cytodifferentiation in stage VIII. The mechanism of chromatin condensation in the elongating spermatids is an elaborate process that encompasses several biochemical and biological aspects culminating in the deposition of protamine in DNA grooves. The protamination of sperm chromatin involves expression and storage of proteins involved in condensation, removal and degradation of nuclear histones and their replacement by transition proteins and protamine 1, transcriptional silencing and DNA repair, reduction of nuclear volume, repackaging of protaminated chromatin in torroids and development of a characteristic head shape and perforatorium. A study was undertaken in my laboratory to delineate the role of follicle stimulating hormone (FSH) and testosterone in the condensation of nuclear chromatin in the elongating spermatids of sexually competent species of rat. Towards this end, sexually competent male Holtzmann rats were treated with 20 mg/Kg/d per os (oral supplementum) of cyproterone acetate and 3 mg/Kg/d i.p (intra peritoneal) of fluphenazine decanoate to induce a functional deficiency in either testosterone or FSH. In both rat models, membrane-impermeable CMA3 fluorescent dye uptake assay for GC-rich prospective sites of DNA protamination, was indicative of insufficiency of protamine 1 in spermatozoa taken from caput epididymides of treated rats whereas a fluorescent TUNEL assay indicated the presence of nicked chromatin strands only in protamine-deficient spermatozoa derived from caput epididymides of fluphenazine-treated rats with functional deficiency of FSH. Western blotting of acid-soluble sperm basic proteins had confirmed the near absence of protamine 1 in treated rat spermatozoa in both models. Electron Microscopic evaluation too revealed fine ultrastructural changes in the nuclear membrane of cyproterone acetate as well as fluphenazine decanoate treated spermatozoa derived from caput epididymides. Electrophoretic analysis of caput sperm nuclear basic proteins substantiated the observations at cellular level and revealed a pattern of abnormal persistence of acid-soluble nuclear basic proteins in both rat models, the levels being more prominent in fluphenazine treated rats. Our studies suggest that adequate levels of both FSH and testosterone could be essential during the stages of spermatidal condensation and led us to hypothesize the existence of an endocrine-regulated molecular mechanism for histone to protamine transition and maintenance of chromatin integrity during chromatin condensation in the testis during spermiogenesis.
authors: Department of Neuroendocrinology, National Institute for Research in Reproductive Health, Parel, Mumbai, India. manjitgill_sharma@rediffmail.com
authors: Manjeet Kaur
authors: MK
authors: Gill-Sharma
authors: Jyoti
authors: J
authors: Choudhuri
authors: Serena
authors: S
authors: D'Souza
mesh_terms: D000818:Animals; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects / Q000502:physiology; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D008297:Male; D011479:Protamines / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013077:Sperm Head / Q000187:drug effects / Q000648:ultrastructure; D013087:Spermatids; D013091:Spermatogenesis / Q000187:drug effects / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism*; D013739:Testosterone / Q000494:pharmacology
chemical_list: D002843:Chromatin; D011479:Protamines; D013739:Testosterone; D005640:Follicle Stimulating Hormone