Changes in the expression of connexin 43, a cardiac gap junctional protein, during mouse heart development.
A cDNA probe coding for rat connexin 43 (Beyer et al., 1987), a gap junctional protein, was used to detect specific mRNA and estimate its relative abundance in mouse heart at different developmental stages: 11, 14 and 19 days post-coïtum (dpc); 1, 2 and 3 weeks post-partum (wpp), and at the adult stage. On Northern blots of total cellular RNA, a single 3.0 kb message was detected at all stages of development, and the differential intensities of labeling indicated developmental changes in mRNA abundance. mRNA levels were further investigated by dot-blotting. Densitometric analyses of dot-blot autoradiograms showed a five-fold increase of the mRNA level between 11 dpc and 1 wpp, then a gradual decrease until the adult stage where it reached a value close to that detected at 11 dpc. By comparison, myosin heavy chains and glycerldehyde-3-phosphate dehydrogenase mRNAs were found to peak at 3 wpp and 14 dpc, respectively. The presence and the relative abundance of connexin 43 were investigated at the same developmental stages as previously by immunoblotting of whole-ventricle fractions using antipeptide antibodies specific for this junctional protein. Quantitative data obtained from densitometric analyses of immunoblots showed that from 14 dpc to 1 wpp intensity of labeling of connexin 43 was roughly multiplied by a factor of 10. It peaked at 3 wpp before dropping to about 20% at the adult stage. The data obtained with both the cDNA probe and the antibodies were significant as shown by variance analyses. They suggest that expression of cardiac connexin 43 is developmentally-regulated: at the early stages of heart development the expression levels of the protein would seem to be mainly regulated by mRNA abundance; beyond 2 weeks after birth, the levels of connexin 43 would seem rather to depend upon its stability and/or the efficiency of the translation.
Fromaget
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Laboratoire de Biologie de la Différenciation Cellulaire, URA CNRS 179, Faculté des Sciences de Luminy, Université d'Aix-Marseille II, France.
el Aoumari
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A
Dupont
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Briand
J P
JP
Gros
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D
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D017630:Connexins; D005786:Gene Expression Regulation; D006321:Heart / Q000196:embryology / Q000254:growth & development*; D007365:Intercellular Junctions / Q000737:chemistry; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism
D017630:Connexins; D008565:Membrane Proteins; D012333:RNA, Messenger