Regulation of desmocollin transcription in mouse preimplantation embryos.
The molecular mechanisms regulating the biogenesis of the first desmosomes to form during mouse embryogenesis have been studied. A sensitive modification of a reverse transcriptase-cDNA amplification procedure has been used to detect transcripts of the desmosomal adhesive cadherin, desmocollin. Sequencing of cDNA amplification products confirmed that two splice variants, a and b, of the DSC2 gene are transcribed coordinately. Transcripts were identified in unfertilized eggs and cumulus cells and in cleavage stages up to the early 8-cell stage, were never detected in compact 8-cell embryos, but were evident again either from the 16-cell morula or very early blastocyst (approx 32-cells) stages onwards. These two phases of transcript detection indicate DSC2 is encoded by maternal and embryonic genomes. Previously, we have shown that desmocollin protein synthesis is undetectable in eggs and cleavage stages but initiates at the early blastocyst stage when desmocollin localises at, and appears to regulate assembly of, nascent desmosomes that form in the trophectoderm but not in the inner cell mass (Fleming, T. P., Garrod, D. R. and Elsmore, A. J. (1991), Development 112, 527-539). Maternal DSC2 mRNA is therefore not translated and presumably is inherited by blastomeres before complete degradation. Our results suggest, however, that initiation of embryonic DSC2 transcription regulates desmocollin protein expression and thereby desmosome formation. Moreover, data from blastocyst single cell analyses suggest that embryonic DSC2 transcription is specific to the trophectoderm lineage. Inhibition of E-cadherin-mediated cell-cell adhesion did not influence the timing of DSC2 embryonic transcription and protein expression. However, isolation and culture of inner cell masses induced an increase in the amount of DSC2 mRNA and protein detected. Taken together, these results suggest that the presence of a contact-free cell surface activates DSC2 transcription in the mouse early embryo.
Collins
J E
JE
Department of Biology, School of Biological Sciences, University of Southampton, UK.
Lorimer
J E
JE
Garrod
D R
DR
Pidsley
S C
SC
Buxton
R S
RS
Fleming
T P
TP
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000502:physiology*; D015820:Cadherins / Q000235:genetics; D004270:DNA, Circular / Q000032:analysis*; D051187:Desmocollins; D003896:Desmosomes / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D007536:Isomerism; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase; D014158:Transcription, Genetic
D015820:Cadherins; D004270:DNA, Circular; D051187:Desmocollins; C489912:Dsc2 protein, mouse; D008562:Membrane Glycoproteins; D012194:RNA-Directed DNA Polymerase