Construction of the nuclear matrix at the transition from maternal to zygotic control of development in the mouse: An immunocytochemical study

Prather, Randall S.; Schatten, Gerald

doi:10.1002/mrd.1080320304

Cited by 20 publications

(11 citation statements)

References 32 publications

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“…Somatic histone H1 is not detected in early mouse embryos until the four-cell stage, in which its synthesis appears to be associated with progression through the S phase of the cell cycle (10). Data concerning the nuclear matrix in preimplantation embryos are extremely sparse, but immunocytochemical evidence suggests that a matrix is formed as early as the two-cell stage in mouse embryos (52). Taken together with these data, our results lend (Fig.…”

Section: Inspection Of the Results For Sar-lines Insupporting

confidence: 83%

Scaffold attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues.

Thompson¹,

Christians²,

Stinnakre³

et al. 1994

Mol. Cell. Biol.

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Eukaryotic interphase chromatin is thought to be organized into topologically discrete, independent domains acting as units upon which differential patterns of gene expression are established. Sequences which attach chromatin to in vitro preparations of a nucleoprotein matrix (scaffold attachment regions [SARs]) may act as domain boundaries, but their role remains poorly defined compared with those of other elements such as locus control regions. We have produced mice homozygous for a transgene which is transcribed as early as the activation of the embryonic genome at the two-cell stage and which is expressed ubiquitously in a number of differentiated tissues. Transgenic lines were generated in the presence or absence of flanking SAR sequences, creating an original model which enabled us to examine the effects of these elements at different developmental stages. In the preimplantation mouse embryo, flanking SARs stimulated transgene expression in a copydependent manner. In contrast, in the differentiated tissues of newborn and adult mice, no significant SAR-dependent increase in transgene expression was found, correlation with copy number was lost, and position effects were observed. These results suggest a limited capacity of SARs to act as insulating elements but are consistent with a proposed model of SAR-mediated chromatin opening and closing.There is increasing evidence that eukaryotic interphase chromatin is arranged on some form of structural skeleton which is thought to play roles in DNA transcription and replication (4,5,7,21,22,34,60). Thus, the eukaryotic genome seems to be divided into topologically constrained domains which may be involved in the organization of differential patterns of gene expression. The boundaries of these domains contain stretches of DNA which bind to in vitro preparations of the nuclear matrix or scaffold. These sequences, termed matrix. (11) (63) proposed an interesting model in which SARs might be involved in chromatin opening via interactions with nuclear proteins. That SAR elements are likely to be implicated in nuclear processes is also supported by the evolutionary conservation of these sequences from yeasts to humans (1,6,28,36,45,50

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Section: Inspection Of the Results For Sar-lines Insupporting

confidence: 83%

Scaffold attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues.

Thompson¹,

Christians²,

Stinnakre³

et al. 1994

Mol. Cell. Biol.

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“…The changes in 1CG organization are consistent with the EM localization of Sm and other snRNP antigens primarily in association with ICG and perichromatin fibrils [34,50,72]. Although the antigen detected by the PII antibody in the nucleoplasm is unknown, it is widely conserved [19,66], and its behaviour during lymphocyte activation [21,69], early embryogenesis [61] and mitosis [17] indicates that it carries out a transcription-related function. EM immunoperoxidase and immunogold labelling studies of 3T3 [22] and Heka (Chert and Cha[y, in preparation) cells have shown that, like snRNP antigens, the internal Pil component is associated with ICG clusters.…”

Section: Nuclear Matrixsupporting

confidence: 68%

Reversible disassembly of transcription domains in lymphocyte nuclei during inhibition of RNA synthesis by DRB

Davis

Cadrin

Brown

et al. 1993

Biology of the Cell

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We are investigating the roles of RNA synthesis, chromatin structure and nuclear matrix organization in establishing and maintaining transcription domains, using mitogen stimulated lymphocytes as a model system. In a continuing study, the effects of the RNA polymerase inhibitor DRB and of its removal on nuclear organization have been examined by EM cytochemistry and by immunofluorescence labelling of the nuclear matrix PI1, Sm and nucleolar fibrillarin antigens. Chromatin, interchromatin granules and nucleoli were extensively restructured after DRB, as were matrix antigens. According to cytochemical staining properties, the conformation of DRB-induced condensed chromatin resembled that in partially stimulated lymphocytes. The nucleoplasmic fibrogranular RNP network appeared little altered, but the fibrillar proteinaceous interchromatinic regions, interpreted as representing the nuclear matrix in situ, were more affected. After removal of DRB, nuclei recovered the organization and transcriptional activity of controls within 8 h. These results suggest that the matrix subtending transcription domains remains stable when transcription is arrested, even though the chromatin and individual RNP components of the domains are disorganized. The data further indicate that absence of transcription is not solely accountable for the highly aggregated state of the chromatin in resting lymphocytes.

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“…Specifically, cycloheximide did not block the appearance of a specific snRNP epitope (Y12), while a-amanitin did block this appearance . Additional data (Prather and Schatten, unpublished) involving the construction of the nuclear matrix of the mouse embryo also showed that long-term treatment with cycloheximide did not block the appearance of some of the nuclear matrix antigens, while a-amanitin did block the developmental appearance of these antigens (Prather and Schatten, 1992). Thus, it appears that these developmental events were not blocked by cycloheximide, because the cycloheximide may no longer have been effective.…”

Section: Discussionmentioning

confidence: 97%

Differential effects of protein synthesis inhibitors on porcine oocyte activation

Nussbaum

Prather

1995

Molecular Reproduction Devel

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This study was designed to evaluate the effects of cycloheximide and puromycin on activation and protein synthesis of porcine oocytes. When matured oocytes were electrostimulated, then cultured in the presence of cycloheximide (5 micrograms/ml) for 6 or 24 hr, 92% of oocytes were activated as indicated by pronuclear formation, vs. 2.8% for untreated oocytes, 5.3% for oocytes not electrostimulated but cultured with cycloheximide, and 60.0% for those only electrostimulated. When cultured with L-[35S]methionine in the presence of cycloheximide, puromycin (100 micrograms/ml), or no protein synthesis inhibitor for 24 hr, oocytes had mean radiolabeled incorporation rates of 36.5, 2.21, and 32.0 fmol/4 hr/oocyte, respectively. Thus, cycloheximide had little effect on protein synthesis after 24 hr of culture. A 1D-SDS PAGE showed that oocytes cultured with puromycin or cycloheximide are not activated, while electrostimulated oocytes are activated, as characterized by the conversion of a 25-kDa polypeptide to a 22-kDa polypeptide. The radiolabeling experiment was repeated, except that oocytes were cultured for 4 or 24 hr. At 4 hr, mean incorporation rates were lower in the cycloheximide group (2.34 fmol/4 hr/oocyte), but similar in the puromycin (15.7 fmol/4 hr/oocyte) and control groups (18.9 fmol/4 hr/oocyte). At 24 hr, the puromycin group 5.73 fmol/4 hr/oocyte) had a lower rate of incorporation, while the cycloheximide (22.6 fmol/4 hr/oocyte) and control (26.0 fmol/4 hr/oocyte) groups were similar. Cycloheximide was more effective earlier during culture, while puromycin was more effective later. When combined with ES, puromycin did have a higher rate (P = 0.10) of activation (87.8%) than with electrostimulation alone (73.0%).(ABSTRACT TRUNCATED AT 250 WORDS)

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Construction of the nuclear matrix at the transition from maternal to zygotic control of development in the mouse: An immunocytochemical study

Cited by 20 publications

References 32 publications

Scaffold attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues.

Scaffold attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues.

Reversible disassembly of transcription domains in lymphocyte nuclei during inhibition of RNA synthesis by DRB

Differential effects of protein synthesis inhibitors on porcine oocyte activation

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