Primordial germ cell deficiency in the connexin 43 knockout mouse arises from apoptosis associated with abnormal p53 activation.
Connexin 43 knockout (Cx43alpha1KO) mice exhibit germ cell deficiency, but the underlying cause for the germ cell defect was unknown. Using an Oct4-GFP reporter transgene, we tracked the distribution and migration of primordial germ cells (PGCs) in the Cx43alpha1KO mouse embryo. Analysis with dye injections showed PGCs are gap-junction-communication competent, with dye coupling being markedly reduced in Cx43alpha1-deficient PGCs. Time-lapse videomicroscopy and motion analysis showed that the directionality and speed of cell motility were reduced in the Cx43alpha1KO PGCs. This was observed both in E8.5 and E11.5 embryos. By contrast, PGC abundance did not differ between wild-type and heterozygous/homozygous Cx43alpha1KO embryos until E11.5, when a marked reduction in PGC abundance was detected in the homozygous Cx43alpha1KO embryos. This was accompanied by increased PGC apoptosis and increased expression of activated p53. Injection of alpha-pifithrin, a p53 antagonist, inhibited PGC apoptosis and prevented the loss of PGC. Analysis using a cell adhesion assay indicated a reduction in beta1-integrin function in the Cx43alpha1KO PGCs. Together with the abnormal activation of p53, these findings suggest the possibility of anoikis-mediated apoptosis. Overall, these findings show Cx43alpha1 is essential for PGC survival, with abnormal p53 activation playing a crucial role in the apoptotic loss of PGCs in the Cx43alpha1KO mouse embryos.
Francis
Richard J B
RJ
Laboratory of Developmental Biology, National Heart Lung and Blood Institute, National Institutes of Health, Building 50/Room 4537, 9000 Rockville Pike, Bethesda, MD 20892, USA.
Lo
Cecilia W
CW
D000482:Allantois / Q000166:cytology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D052160:Benzothiazoles / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology*; D018031:Connexin 43 / Q000172:deficiency / Q000502:physiology*; D005260:Female; D017930:Genes, Reporter; D016158:Genes, p53 / Q000502:physiology*; D005838:Genotype; D005854:Germ Cells / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D014050:Toluene / Q000031:analogs & derivatives / Q000494:pharmacology; D019076:Transgenes
D052160:Benzothiazoles; D018031:Connexin 43; D014050:Toluene; C121565:pifithrin