Differential regulation of multiple gap junction transcripts and proteins during rat liver regeneration

Betsy T. Kren, Nalin M. Kumar, Sheng Qi Wang, Norton B. Gilula, Clifford J Steer

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The mRNA and protein expression of α1 (connexin 43), β1 (connexin 32), and β2 (connexin 26) gap junction genes were examined in the regenerating rat liver after 70% partial hepatectomy (PH). Expression of β1 and β2 steady-state mRNA levels changed minimally until 12 h after PH when both transcripts decreased to approximately 15% of baseline values. A similar decrease in assembled connexin levels was detected by immunoblot and indirect immunofluorescence at 18 h after PH. Both transcripts simultaneously increased between 24 and 42 h and again rapidly decreased by 48 h post-PH. β1 and β2 assembled gap junction protein expression increased at 48 h post-PH and rapidly decreased by 56 h. By 72 to 84 h post-PH, β1 and β2 mRNA and assembled protein expression returned to near baseline levels and were maintained. Interestingly, inhibition of protein synthesis with cycloheximide completely inhibited disappearance of the β2 transcript, in contrast to β1 mRNA which was unaffected. Nuclear run-on assays showed no change in transcriptional rates for either gene during the regenerative period. However, both β1 and β2 transcripts exhibited significantly decreased mRNA half-lives at 12 h post-PH (3.8 and 3.7 h, respectively) relative to those at 0 h (10.9 and 6.1 h, respectively). Surprisingly, although the transcriptional rate for α1 was similar to that observed for β2, no α1, transcripts were detectable by northern or RNase protection analysis. The results suggest that in the regenerating rat liver, β1 and β2 gap junction genes are not regulated at the transcriptional level. Rather, the cyclical modulation of their steady-state transcripts is regulated primarily by posttranscriptional events of which mRNA stability is at least one critical factor in the control process.

Original languageEnglish (US)
Pages (from-to)707-718
Number of pages12
JournalJournal of Cell Biology
Issue number3
StatePublished - Nov 1993


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