Mu-opioid receptor mRNA regulation during morphine tolerance in the rat peripheral nervous system.
In vivo data on opioid receptor mRNA regulation after agonist exposure in the peripheral nervous system are lacking. Therefore, we studied the impact of morphine treatment on the regulation of mu-opioid receptor mRNA during behavioral signs of tolerance in rat peripheral sensory ganglia. Nineteen rats were treated in 2 groups with either morphine (10 mg/kg subcutaneously) or saline over 4 days, and a subset of rats received naloxone on the fifth day followed by either morphine injection on the sixth day or death to obtain dorsal root ganglia for mRNA analysis. Animals were tested on the hot plate during treatment days. To assess the levels of mu-opioid receptor mRNA, quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used with the co-amplification of the "housekeeping" gene cyclophilin as internal control. Morphine treatment over 4 days induced tolerance as reflected on the hot-plate test by a significant reduction of paw-withdrawal latency from 242% to 99% above baseline. Using RT-PCR we demonstrated a down-regulation of mu-opioid receptor mRNA by 62% after morphine exposure (P < 0.05). After acute withdrawal of morphine from the mu-receptor by naloxone, the mu-opioid receptor mRNA levels in the dorsal root ganglia were restored to control levels within 24 h and the paw-withdrawal latency also returned to 280% above control. These data suggest that the peripheral nervous system may be an important site of opioid tolerance development.


CONCLUSIONS
The peripheral nervous system is a possible site of opioid receptor tolerance. We show the development of behavioral tolerance and mu-opioid receptor mRNA down-regulation in the dorsal root ganglia in rats after chronic morphine treatment. Both this mRNA down-regulation and behavioral tolerance reverse after 24 h of naloxone treatment.
Departments of *Anesthesiology and †Neurophysiology, University of Cologne, Cologne, Germany, ‡Department of Anesthesiology, University of Halle Wittenberg, Halle, Germany, and the §Department of Anesthesia, University of California, San Francisco, CA.
Thomas
T
Meuser
Thorsten
T
Giesecke
Anja
A
Gabriel
Maria
M
Horsch
Rainer
R
Sabatowski
Jürgen
J
Hescheler
Stefan
S
Grond
Pamela Pierce
PP
Palmer
D000701:Analgesics, Opioid / Q000494:pharmacology*; D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects; D004361:Drug Tolerance; D004587:Electrophoresis, Agar Gel; D006358:Hot Temperature; D008297:Male; D009020:Morphine / Q000494:pharmacology*; D009270:Naloxone / Q000494:pharmacology; D009292:Narcotic Antagonists / Q000494:pharmacology; D010147:Pain Measurement / Q000187:drug effects; D017933:Peripheral Nervous System / Q000187:drug effects*; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011930:Reaction Time / Q000187:drug effects; D017450:Receptors, Opioid, mu / Q000096:biosynthesis* / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000701:Analgesics, Opioid; D009292:Narcotic Antagonists; D012333:RNA, Messenger; D017450:Receptors, Opioid, mu; D009270:Naloxone; D009020:Morphine