Immunofluorescent localization of actin in relation to transcription sites in mouse pronuclei.
Previous biochemical and morphological studies have shown the presence of actin in the nucleus of different cell types where its role remains unclear. In this work, through fluorescence microscopy we studied the localization of actin in the nuclei of early mouse embryos with particular attention to its possible involvement in the onset of transcription occurring at the late one-cell stage. Fluorescent labelling of embryo sections showed that nuclear actin in abundant, in a non-filamentous state, in the whole nucleoplasm excluding the nucleolar precursor bodies. Immunofluorescence on permeabilized embryos revealed that insoluble nuclear actin accumulates in a few large aggregates in transcriptionally inert early one-cell embryos and progressively redistributes into many small aggregates in transcriptionally active late one-cell embryos. Interestingly, these actin aggregates clearly colocalize with transcription sites. Treatment of late one-cell embryos with cytochalasin D induces the formation of actin bundles network in the nucleoplasm but has no apparent effect on the transcriptional activity. In addition, the inhibition of transcription by alpha-amanitin does not modify the nuclear actin distribution. Hence, there does not appear to be a direct causal relationship between transcriptional activity and nuclear actin organization at the one-cell stage although nuclear actin aggregates appear associated with transcription sites.
Nguyen
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Laboratoire Associé INSERM (Unité 310), INRA (Station 806) Institut de Biologie Physico-Chimique, Paris, France. nguyen@ibpc.fr
Besombes
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Debey
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D000199:Actins / Q000378:metabolism*; D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D015638:Cytochalasin D / Q000494:pharmacology; D004622:Embryo, Mammalian; D005314:Embryonic and Fetal Development; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008856:Microscopy, Fluorescence; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D014158:Transcription, Genetic
D000199:Actins; D000546:Amanitins; D019384:Nucleic Acid Synthesis Inhibitors; D015638:Cytochalasin D