Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Jul;96(1):231–238. doi: 10.1172/JCI118026

Stress-induced activation of neuronal activity and corticotropin-releasing factor gene expression in the paraventricular nucleus is modulated by glucocorticoids in rats.

T Imaki 1, W Xiao-Quan 1, T Shibasaki 1, K Yamada 1, S Harada 1, N Chikada 1, M Naruse 1, H Demura 1
PMCID: PMC185193  PMID: 7615792

Abstract

Intronic in situ hybridization methodology provides a means of determining the rate of gene transcription under basal and stimulated conditions. In the present study, we have used intronic in situ hybridization to the corticotropin-releasing factor (CRF) gene to measure hypothalamic CRF gene transcription after stress as well as its modulation by glucocorticoids. Using this and conventional exonic in situ hybridization we examined the time course of changes in c-fos mRNA, and CRF heteronuclear RNA (hnRNA) and mRNA concentrations in the paraventricular nucleus (PVN) of male Wistar rats after restraint stress. In addition, we determined the effects of adrenalectomy and dexamethasone administration on c-fos and CRF gene expression in the PVN. Restraint stress induced a rapid induction (within 5 min) of c-fos mRNA and CRF hnRNA expression in the PVN. Both RNA concentrations peaked at 30 min then decreased and were undetectable 2 h after stress onset. In contrast, the concentration of CRF mRNA increased gradually and a significant elevation was first detected 60 min after the beginning of stress. Adrenalectomy augmented and dexamethasone pretreatment inhibited c-fos mRNA, CRF hnRNA, and mRNA induction after stress. The data suggest that stress-induced activation of neurons, CRF gene transcription, and CRF synthesis in the PVN are modulated by glucocorticoids.

Full text

PDF
231

Images in this article

233Image
on p.233
233Image
on p.233
234Image
on p.234
234Image
on p.234

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adler G. K., Rosen L. B., Fiandaca M. J., Majzoub J. A. Protein kinase-C activation increases the quantity and poly(A) tail length of corticotropin-releasing hormone messenger RNA in NPLC cells. Mol Endocrinol. 1992 Mar;6(3):476–484. doi: 10.1210/mend.6.3.1350054. [DOI] [PubMed] [Google Scholar]
  2. Adler G. K., Smas C. M., Majzoub J. A. Expression and dexamethasone regulation of the human corticotropin-releasing hormone gene in a mouse anterior pituitary cell line. J Biol Chem. 1988 Apr 25;263(12):5846–5852. [PubMed] [Google Scholar]
  3. Autelitano D. J., Blum M., Roberts J. L. Changes in rat pituitary nuclear and cytoplasmic pro-opiomelanocortin RNAs associated with adrenalectomy and glucocorticoid replacement. Mol Cell Endocrinol. 1989 Oct;66(2):171–180. doi: 10.1016/0303-7207(89)90029-4. [DOI] [PubMed] [Google Scholar]
  4. Carrazana E. J., Pasieka K. B., Majzoub J. A. The vasopressin mRNA poly(A) tract is unusually long and increases during stimulation of vasopressin gene expression in vivo. Mol Cell Biol. 1988 Jun;8(6):2267–2274. doi: 10.1128/mcb.8.6.2267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ceccatelli S., Villar M. J., Goldstein M., Hökfelt T. Expression of c-Fos immunoreactivity in transmitter-characterized neurons after stress. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9569–9573. doi: 10.1073/pnas.86.23.9569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coveñas R., de León M., Cintra A., Bjelke B., Gustafsson J. A., Fuxe K. Coexistence of c-Fos and glucocorticoid receptor immunoreactivities in the CRF immunoreactive neurons of the paraventricular hypothalamic nucleus of the rat after acute immobilization stress. Neurosci Lett. 1993 Jan 12;149(2):149–152. doi: 10.1016/0304-3940(93)90758-d. [DOI] [PubMed] [Google Scholar]
  7. Darnell J. E., Jr The processing of RNA. Sci Am. 1983 Oct;249(4):90–100. doi: 10.1038/scientificamerican1083-90. [DOI] [PubMed] [Google Scholar]
  8. Dragunow M., Faull R. The use of c-fos as a metabolic marker in neuronal pathway tracing. J Neurosci Methods. 1989 Sep;29(3):261–265. doi: 10.1016/0165-0270(89)90150-7. [DOI] [PubMed] [Google Scholar]
  9. Emanuel R. L., Girard D. M., Thull D. L., Majzoub J. A. Second messengers involved in the regulation of corticotropin-releasing hormone mRNA and peptide in cultured rat fetal hypothalamic primary cultures. Endocrinology. 1990 Jun;126(6):3016–3021. doi: 10.1210/endo-126-6-3016. [DOI] [PubMed] [Google Scholar]
  10. Fink G., Robinson I. C., Tannahill L. A. Effects of adrenalectomy and glucocorticoids on the peptides CRF-41, AVP and oxytocin in rat hypophysial portal blood. J Physiol. 1988 Jul;401:329–345. doi: 10.1113/jphysiol.1988.sp017165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fremeau R. T., Jr, Lundblad J. R., Pritchett D. B., Wilcox J. N., Roberts J. L. Regulation of pro-opiomelanocortin gene transcription in individual cell nuclei. Science. 1986 Dec 5;234(4781):1265–1269. doi: 10.1126/science.3775385. [DOI] [PubMed] [Google Scholar]
  12. Harbuz M. S., Lightman S. L. Glucocorticoid inhibition of stress-induced changes in hypothalamic corticotrophin-releasing factor messenger RNA and proenkephalin A messenger RNA. Neuropeptides. 1989 Jul;14(1):17–20. doi: 10.1016/0143-4179(89)90029-2. [DOI] [PubMed] [Google Scholar]
  13. Harbuz M. S., Lightman S. L. Responses of hypothalamic and pituitary mRNA to physical and psychological stress in the rat. J Endocrinol. 1989 Sep;122(3):705–711. doi: 10.1677/joe.0.1220705. [DOI] [PubMed] [Google Scholar]
  14. Harbuz M. S., Nicholson S. A., Gillham B., Lightman S. L. Stress responsiveness of hypothalamic corticotrophin-releasing factor and pituitary pro-opiomelanocortin mRNAs following high-dose glucocorticoid treatment and withdrawal in the rat. J Endocrinol. 1990 Dec;127(3):407–415. doi: 10.1677/joe.0.1270407. [DOI] [PubMed] [Google Scholar]
  15. Herman J. P., Schafer M. K., Thompson R. C., Watson S. J. Rapid regulation of corticotropin-releasing hormone gene transcription in vivo. Mol Endocrinol. 1992 Jul;6(7):1061–1069. doi: 10.1210/mend.6.7.1324419. [DOI] [PubMed] [Google Scholar]
  16. Herman J. P., Schäfer M. K., Watson S. J., Sherman T. G. In situ hybridization analysis of arginine vasopressin gene transcription using intron-specific probes. Mol Endocrinol. 1991 Oct;5(10):1447–1456. doi: 10.1210/mend-5-10-1447. [DOI] [PubMed] [Google Scholar]
  17. Imaki T., Nahan J. L., Rivier C., Sawchenko P. E., Vale W. Differential regulation of corticotropin-releasing factor mRNA in rat brain regions by glucocorticoids and stress. J Neurosci. 1991 Mar;11(3):585–599. doi: 10.1523/JNEUROSCI.11-03-00585.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Imaki T., Nahon J. L., Sawchenko P. E., Vale W. Widespread expression of corticotropin-releasing factor messenger RNA and immunoreactivity in the rat olfactory bulb. Brain Res. 1989 Sep 4;496(1-2):35–44. doi: 10.1016/0006-8993(89)91050-0. [DOI] [PubMed] [Google Scholar]
  19. Imaki T., Shibasaki T., Hotta M., Demura H. Early induction of c-fos precedes increased expression of corticotropin-releasing factor messenger ribonucleic acid in the paraventricular nucleus after immobilization stress. Endocrinology. 1992 Jul;131(1):240–246. doi: 10.1210/endo.131.1.1612001. [DOI] [PubMed] [Google Scholar]
  20. Jingami H., Matsukura S., Numa S., Imura H. Effects of adrenalectomy and dexamethasone administration on the level of prepro-corticotropin-releasing factor messenger ribonucleic acid (mRNA) in the hypothalamus and adrenocorticotropin/beta-lipotropin precursor mRNA in the pituitary in rats. Endocrinology. 1985 Oct;117(4):1314–1320. doi: 10.1210/endo-117-4-1314. [DOI] [PubMed] [Google Scholar]
  21. Lightman S. L., Young W. S., 3rd Corticotrophin-releasing factor, vasopressin and pro-opiomelanocortin mRNA responses to stress and opiates in the rat. J Physiol. 1988 Sep;403:511–523. doi: 10.1113/jphysiol.1988.sp017261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lightman S. L., Young W. S., 3rd Influence of steroids on the hypothalamic corticotropin-releasing factor and preproenkephalin mRNA responses to stress. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4306–4310. doi: 10.1073/pnas.86.11.4306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Minami S., Kamegai J., Sugihara H., Hasegawa O., Wakabayashi I. Systemic administration of recombinant human growth hormone induces expression of the c-fos gene in the hypothalamic arcuate and periventricular nuclei in hypophysectomized rats. Endocrinology. 1992 Jul;131(1):247–253. doi: 10.1210/endo.131.1.1612002. [DOI] [PubMed] [Google Scholar]
  24. Morgan J. I., Curran T. Stimulus-transcription coupling in the nervous system: involvement of the inducible proto-oncogenes fos and jun. Annu Rev Neurosci. 1991;14:421–451. doi: 10.1146/annurev.ne.14.030191.002225. [DOI] [PubMed] [Google Scholar]
  25. Owens M. J., Nemeroff C. B. Physiology and pharmacology of corticotropin-releasing factor. Pharmacol Rev. 1991 Dec;43(4):425–473. [PubMed] [Google Scholar]
  26. Paek I., Axel R. Glucocorticoids enhance stability of human growth hormone mRNA. Mol Cell Biol. 1987 Apr;7(4):1496–1507. doi: 10.1128/mcb.7.4.1496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Priou A., Oliver C., Grino M. In situ hybridization of arginine vasopressin (AVP) heteronuclear ribonucleic acid reveals increased AVP gene transcription in the rat hypothalamic paraventricular nucleus in response to emotional stress. Acta Endocrinol (Copenh) 1993 May;128(5):466–472. doi: 10.1530/acta.0.1280466. [DOI] [PubMed] [Google Scholar]
  28. Rivier C. L., Plotsky P. M. Mediation by corticotropin releasing factor (CRF) of adenohypophysial hormone secretion. Annu Rev Physiol. 1986;48:475–494. doi: 10.1146/annurev.ph.48.030186.002355. [DOI] [PubMed] [Google Scholar]
  29. Rivier C., Rivier J., Vale W. Inhibition of adrenocorticotropic hormone secretion in the rat by immunoneutralization of corticotropin-releasing factor. Science. 1982 Oct 22;218(4570):377–379. doi: 10.1126/science.6289439. [DOI] [PubMed] [Google Scholar]
  30. Rosen L. B., Majzoub J. A., Adler G. K. Effects of glucocorticoid on corticotropin-releasing hormone gene regulation by second messenger pathways in NPLC and AtT-20 cells. Endocrinology. 1992 Apr;130(4):2237–2244. doi: 10.1210/endo.130.4.1547737. [DOI] [PubMed] [Google Scholar]
  31. Sagar S. M., Sharp F. R., Curran T. Expression of c-fos protein in brain: metabolic mapping at the cellular level. Science. 1988 Jun 3;240(4857):1328–1331. doi: 10.1126/science.3131879. [DOI] [PubMed] [Google Scholar]
  32. Seasholtz A. F., Thompson R. C., Douglass J. O. Identification of a cyclic adenosine monophosphate-responsive element in the rat corticotropin-releasing hormone gene. Mol Endocrinol. 1988 Dec;2(12):1311–1319. doi: 10.1210/mend-2-12-1311. [DOI] [PubMed] [Google Scholar]
  33. Sharp F. R., Sagar S. M., Hicks K., Lowenstein D., Hisanaga K. c-fos mRNA, Fos, and Fos-related antigen induction by hypertonic saline and stress. J Neurosci. 1991 Aug;11(8):2321–2331. doi: 10.1523/JNEUROSCI.11-08-02321.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sternberg E. M., Young W. S., 3rd, Bernardini R., Calogero A. E., Chrousos G. P., Gold P. W., Wilder R. L. A central nervous system defect in biosynthesis of corticotropin-releasing hormone is associated with susceptibility to streptococcal cell wall-induced arthritis in Lewis rats. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4771–4775. doi: 10.1073/pnas.86.12.4771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thompson R. C., Seasholtz A. F., Herbert E. Rat corticotropin-releasing hormone gene: sequence and tissue-specific expression. Mol Endocrinol. 1987 May;1(5):363–370. doi: 10.1210/mend-1-5-363. [DOI] [PubMed] [Google Scholar]
  36. Vale W., Rivier C., Brown M. R., Spiess J., Koob G., Swanson L., Bilezikjian L., Bloom F., Rivier J. Chemical and biological characterization of corticotropin releasing factor. Recent Prog Horm Res. 1983;39:245–270. doi: 10.1016/b978-0-12-571139-5.50010-0. [DOI] [PubMed] [Google Scholar]
  37. Vamvakopoulos N. C., Chrousos G. P. Regulated activity of the distal promoter-like element of the human corticotropin-releasing hormone gene and secondary structural features of its corresponding transcripts. Mol Cell Endocrinol. 1993 Jul;94(1):73–78. doi: 10.1016/0303-7207(93)90053-m. [DOI] [PubMed] [Google Scholar]
  38. Vamvakopoulos N. C., Chrousos G. P. Structural organization of the 5' flanking region of the human corticotropin releasing hormone gene. DNA Seq. 1993;4(3):197–206. doi: 10.3109/10425179309015632. [DOI] [PubMed] [Google Scholar]
  39. Van L. P., Spengler D. H., Holsboer F. Glucocorticoid repression of 3',5'-cyclic-adenosine monophosphate-dependent human corticotropin-releasing-hormone gene promoter activity in a transfected mouse anterior pituitary cell line. Endocrinology. 1990 Sep;127(3):1412–1418. doi: 10.1210/endo-127-3-1412. [DOI] [PubMed] [Google Scholar]
  40. Young W. S., 3rd, Mezey E., Siegel R. E. Quantitative in situ hybridization histochemistry reveals increased levels of corticotropin-releasing factor mRNA after adrenalectomy in rats. Neurosci Lett. 1986 Oct 8;70(2):198–203. doi: 10.1016/0304-3940(86)90463-5. [DOI] [PubMed] [Google Scholar]
  41. Zingg H. H., Lefebvre D. L., Almazan G. Regulation of poly(A) tail size of vasopressin mRNA. J Biol Chem. 1988 Aug 15;263(23):11041–11043. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES