Regulation of both preproenkephalin mRNA and its derived opioids by haloperidol--a method for measurement of peptides and mRNA in the same tissue extract.
The goal of this study was to delineate the effects of dopamine antagonists on the regulation of preproenkephalin mRNA and opioid peptides in the rat brain. We have developed a method whereby both mRNA and peptides can be efficiently measured in the same tissue extract, thus reducing the effects of intraspecies variation, differences in dissection and the number of animals required for statistical significance. A sub-chronic dose of haloperidol (3 mg/kg given i.p. in 100 microliters DMSO daily for 5 days) produced a 1.8-fold increase (P less than 0.001) in striatal preproenkephalin mRNA levels when compared to animals injected with vehicle dimethyl sulfoxide (DMSO) employing the same schedule. Total opioid peptides as measured by a radioimmunoassay directed to the N-terminus of enkephalins and endorphins were elevated 1.6 fold (P less than 0.001) in the rat striatum. However in other brain regions examined no increases were observed either in preproenkephalin mRNA or the tissue levels of opioid peptides. Analysis of the opioid-like immunoreactive peptides by reverse-phase HPLC analysis showed no dramatic changes in the ratios of the various opioid peptides between haloperidol and vehicle injected animals. Naive animals showed no statistical differences in opioid peptide levels compared to the haloperidol treated animals. There was a statistically significant decrease (30%) in the opioid peptide content of the animals injected with vehicle daily for 5 days when compared with the animals merely sacrificed, or those given acute injections (either with haloperidol or vehicle) the day of sacrifice.(ABSTRACT TRUNCATED AT 250 WORDS)
Abood
M E
ME
Department of Psychiatry and Behavioral Sciences, Stanford University Medical Center, CA 94305.
Eberwine
J H
JH
Erdelyi
E
E
Evans
C J
CJ
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D001971:Bromocriptine / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004745:Enkephalins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D005786:Gene Expression Regulation / Q000187:drug effects; D006220:Haloperidol / Q000494:pharmacology*; D008297:Male; D009928:Organ Specificity; D011498:Protein Precursors / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000187:drug effects / Q000235:genetics*; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D004745:Enkephalins; D011498:Protein Precursors; D012333:RNA, Messenger; D001971:Bromocriptine; C033660:preproenkephalin; D006220:Haloperidol