Developmental regulation of chromatin composition during mouse embryogenesis: somatic histone H1 is first detectable at the 4-cell stage.
We have examined the distribution of histone H1 in oocytes and preimplantation embryos of the mouse, using a polyclonal antibody raised against the histone H1 subtypes present in somatic cells. Immunofluorescence and immunoblotting analyses failed to detect somatic histone H1 in germinal vesicle (GV)-stage oocytes. In contrast, somatic histone H1 was detectable by immunofluorescence in the nuclei of GV oocytes previously injected with histone H1 as well as the nuclei of ovarian granulosa cells, and by immunoblotting in 8-cell embryos. 1- and 2-cell embryos examined by immunofluorescence did not contain detectable somatic histone H1. At the early 4-cell stage (54-56 hours post-hCG), 5 of 52 embryos contained somatic histone H1 in one or more nuclei. By the late 4-cell stage (66-68 hours post-hCG), however, 58 of 62 embryos contained somatic histone H1. In 8-cell embryos, morulae and blastocysts, all nuclei contained somatic histone H1 in every case. When embryos were exposed to the transcriptional inhibitor, alpha-amanitin, beginning at the late 2-cell stage, they cleaved to the 4-cell stage but fewer than 10% developed histone H1 immunoreactivity. When treatment began at the early 4-cell stage, the embryos that remained at the 4-cell stage in the presence of the drug developed histone H1 immunoreactivity in half of the cases. Embryos that reached the 5- to 8-cell stage in the presence of the drug developed histone H1 immunoreactivity in every case.(ABSTRACT TRUNCATED AT 250 WORDS)
Clarke
H J
HJ
Department of Obstetrics & Gynecology, McGill University, Montreal, Canada.
Oblin
C
C
Bustin
M
M
D000818:Animals; D001755:Blastocyst / Q000737:chemistry*; D002843:Chromatin / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D006657:Histones / Q000737:chemistry*; D015151:Immunoblotting; D051379:Mice; D009024:Morphogenesis / Q000502:physiology; D009865:Oocytes / Q000737:chemistry*
D002843:Chromatin; D006657:Histones