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 of the mRNAs for alpha(1)-adrenoceptor subtypes, during dissociation of guinea pig liver cells with collagenase. Using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) assays, it was 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 actinomycin 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)-adrenoceptor mRNA expression.
González-Espinosa
C
C
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México Apartado Postal 70-248, Mexico, Mexico.
González-Espinosa
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D
Romero-Avila
M T
MT
García-Sáinz
J A
JA
D000818:Animals; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D005786:Gene Expression Regulation; D006168:Guinea Pigs; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012313:RNA / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018340:Receptors, Adrenergic, alpha-1 / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D018340:Receptors, Adrenergic, alpha-1; D003609:Dactinomycin; D012313:RNA; D003513:Cycloheximide