Abstract
Chromatin is known to undergo extensive remodeling during nuclear reprogramming. However, the factors and mechanisms involved in this remodeling are still poorly understood and current experimental approaches to study it are not best suited for molecular and genetic analyses. Here we report on the use of Drosophila preblastodermic embryo extracts (DREX) in chromatin remodeling experiments. Our results show that incubation of somatic nuclei in DREX induces changes in chromatin organization similar to those associated with nuclear reprogramming, such as rapid binding of the germline specific linker histone dBigH1 variant to somatic chromatin, heterochromatin reorganization, changes in the epigenetic state of chromatin, and nuclear lamin disassembly. These results raise the possibility of using the powerful tools of Drosophila genetics for the analysis of chromatin changes associated with this essential process.
Highlights
Chromatin remodeling is essential for nuclear reprogramming
An early event in reprogramming of somatic nuclei transplanted into oocytes is the binding to chromatin of the oocyte specific linker histones H17,19–22
We report that incubation of somatic nuclei in Drosophila preblastodermic embryo extracts (DREX) induces changes in chromatin organization similar to those associated with nuclear reprogramming
Summary
Chromatin remodeling is essential for nuclear reprogramming. Experimental approaches to study chromatin remodeling during nuclear reprogramming include ectopic expression of Yamanaka transcription factors, nuclear transfer to eggs or oocytes, cell fusion with embryonic stem cells and in vitro treatment with oocyte or egg extracts[1,2,3,4,5,6,7,8,9]. In Drosophila, histone H1 complexity is reduced to a single somatic dH1 variant[26,27,28], and a second germline specific dBigH1 isoform, which is present in the early embryo until activation of the zygotic genome (ZGA) at cellularization[29]. We show that DREX induces heterochromatin reorganization, nuclear lamin disassembly and changes in the pattern of histone modifications, all of which are associated with reprogramming of somatic nuclei. These results suggest that DREX induces partial remodeling of somatic chromatin, opening up the possibility of using the powerful tools of Drosophila genetics to study this central step in somatic cells reprogramming
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