Inverse alpha(1A) and alpha(1D) adrenoceptor mRNA expression during isolation of hepatocytes

It is now well documented that changes in gene expression take place during cell isolation and culture. Here, we report the change in the expression o f the mRNAs for alpha(1)-adrenoceptor subtypes, during dissociation of guin ea pig liver cells with collagenase. Using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) assays, it w as observed that during the isolation procedure, the mRNA for the alpha(1A) -adrenoceptor, normally expressed in whole liver, was degraded and the mRNA for alpha(1D) subtype, barely expressed in whole liver, increased in an ac tinomycin D-sensitive manner. When the isolation procedure was performed in the presence of cycloheximide, the mRNA for the alpha(1A)-adrenoceptor did not diminish and the induction of the alpha(1D)-adrenoceptor mRNA was even more evident. Our data indicate that cell isolation alters alpha(1)-adreno ceptor mRNA expression. (C) 1999 Elsevier Science B.V. All rights reserved.