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Mouse Oocytes and Early Embryos Express Multiple Histone H1 Subtypes¹

Departments of Obstetrics and Gynecology⁵ Biology,⁶ McGill University, Montréal, Québec, Canada H3A 1A1 Department of Cell Biology,⁷ Albert Einstein College of Medicine, Yeshiva University, Bronx, New York 10461 INSERM U309,⁸ Institut Albert Bonniot, Grenoble, France

Oocytes and embryos of many species, including mammals, contain a unique linker (H1) histone, termed H1oo in mammals. It is uncertain, however, whether other H1 histones also contribute to the linker histone complement of these cells. Using immunofluorescence and radiolabeling, we have examined whether histone H1⁰, which frequently accumulates in the chromatin of nondividing cells, and the somatic subtypes of H1 are present in mouse oocytes and early embryos. We report that oocytes and embryos contain mRNA encoding H1⁰. A polymerase chain reaction-based test indicated that the poly(A) tail did not lengthen during meiotic maturation, although it did so beginning at the four-cell stage. Antibodies raised against histone H1⁰ stained the nucleus of wild-type prophase-arrested oocytes but not of mice lacking the H1⁰ gene. Following fertilization, H1⁰ was detected in the nuclei of two-cell embryos and less strongly at the four-cell stage. No signal was detected in H1⁰ -/- embryos. Radiolabeling revealed that species comigrating with the somatic H1 subtypes H1a and H1c were synthesized in maturing oocytes and in one- and two-cell embryos. Beginning at the four-cell stage in both wild-type and H1⁰ -/- embryos, species comigrating with subtypes H1b, H1d, and H1e were additionally synthesized. These results establish that histone H1⁰ constitutes a portion of the linker histone complement in oocytes and early embryos and that changes in the pattern of somatic H1 synthesis occur during early embryonic development. Taken together with previous results, these findings suggest that multiple H1 subtypes are present on oocyte chromatin and that following fertilization changes in the histone H1 complement accompany the establishment of regulated embryonic gene expression.

¹ Supported by operating grants from the Canadian Institutes of Health Research (H.J.C.), NIH grant CA 79057 (A.I.S.), and a fellowship from the Natural Sciences and Engineering Research council (G.F.).

² Correspondence: H.J. Clarke, Room F3.50, Royal Victoria Hospital, 687 av. des Pins O., Montréal, Québec, Canada H3A 1A1. FAX: 514 8431662; hugh.clarke{at}muhc.mcgill.ca

³ Current address: University of California at Santa Barbara, Santa Barbara, CA

⁴ Current address: Department of Biochemistry and Biophysics, Tehran University, Tehran, Iran

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