STRESS RESPONSIVENESS OF HYPOTHALAMIC CORTICOTROPIN-RELEASING FACTOR AND PITUITARY PROOPIOMELANOCORTIN MESSENGER-RNAS FOLLOWING HIGH-DOSE GLUCOCORTICOID TREATMENT AND WITHDRAWAL IN THE RAT

In-situ hybridization with synthetic oligonucleotide probes was used to determine the mRNA content of corticotrophin-releasing factor (CRF) and proenkephalin A mRNA in the paraventricular nucleus, and of pro-opiomelanocortin (POMC) mRNA in the anterior pituitary gland of male rats immediately after, and during recovery from, chronic high-dose prednisolone treatment. Levels of transcripts for mRNA for both CRF and POMC were markedly reduced after the treatment, but there was a rapid return to control values for CRF mRNA within 18 h of steroid withdrawal. In untreated animals, the stressful stimulus of i.p. hypertonic saline increased CRF and proenkephalin A mRNA within 4 h with no significant difference in response seen whether the tissues were removed at 13.00 or 20.00 h. The increase in POMC mRNA did not reach statistical significance in these animals. Although prednisolone resulted in a marked reduction of basal CRF mRNA, the stress-induced increment of CRF mRNA remained comparable with that found in untreated animals. On the day following cessation of prednisolone treatment at 09.00 h, basal and stress levels of CRF mRNA were significantly higher in rats killed at 20.00 h than at 13.00 h. Proenkephalin A mRNA transcripts were below quantifiable levels of detection in control or non-stressed prednisolone-treated animals at all the time-points studied. Stress, however, resulted in the accumulation of proenkephalin A mRNA in control animals. This response was inhibited by prednisolone treatment and only returned 18 h after withdrawal. Prednisolone treatment reduced POMC mRNA below the levels detected in untreated animals, with no detectable response to stress. However, 36 h after prednisolone withdrawal there was a significant increase in POMC mRNA in response to stress. Prednisolone concentrations in the plasma on the morning of the day after the final administration of the steroid were very high (880 nmol/l), but these were halved by the evening of that day and had fallen to 60 nmol/l 72 h after withdrawal. These data show that both CRF and POMC mRNA were profoundly reduced by long-term prednisolone treatment. Stress-induced accumulation of CRF mRNA, however, persisted although the proenkephalin A mRNA responses to this stress were inhibited. No increase in pituitary POMC mRNA in response to stress was apparent until 36 h after withdrawal of prednisolone.