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Volume 17, Number 4, Issue of February 15, 1997 pp. 1302-1319
Copyright ©1997 Society for Neuroscience

Chronic Alcohol Consumption and Withdrawal Do Not Induce Cell Death in the Suprachiasmatic Nucleus, But Lead to Irreversible Depression of Peptide Immunoreactivity and mRNA Levels

Received Sept. 3, 1996; revised Nov. 11, 1996; accepted Dec. 3, 1996.

M. D. Madeira¹, J. P. Andrade¹, A. R. Lieberman², N. Sousa¹, O. F. X. Almeida³, and M. M. Paula-Barbosa¹

¹ Department of Anatomy, Porto Medical School, 4200 Porto, Portugal, ² Department of Anatomy and Developmental Biology, University College London, London WC1E 6BT, United Kingdom, and ³ Max-Planck-Institute of Psychiatry, Clinical Institute, D-W-80804 Munich, Germany

There is evidence that chronic ethanol treatment (CET) disrupts the biological rhythms of various brain functions and behaviors. Because the suprachiasmatic nucleus (SCN) is widely recognized as the dominant pacemaker of the circadian system, we have examined the effects of CET and withdrawal on the main morphological features and chemoarchitecture of this hypothalamic nucleus. Groups of rats ethanol-treated for 6 and 12 months were compared with withdrawn rats (ethanol-treated for 6 months and then switched to a normal diet for an additional 6 months) and with groups of age-matched control and pair-fed control rats. The volume and the total number of neurons of the SCN were estimated from conventionally stained material, whereas the total number of astrocytes and of neurons containing vasopressin (AVP), vasoactive intestinal polypeptide (VIP), gastrin-releasing peptide (GRP), and somatostatin (SS) were estimated from immunostained sections. The estimates were obtained using unbiased stereological methods, based on Cavalieri's principle and the optical fractionator. The volume of the SCN and the total number of SCN neurons and astrocytes did not vary among groups. We found, however, that CET induced a significant reduction in the total number of AVP-, VIP-, GRP-, and SS-containing neurons. Withdrawal from alcohol did not reduce but rather augmented the loss of VIP- and GRP-immunoreactive neurons. The CET-induced neurochemical alterations seem to result from a decrease in neuropeptide synthesis, as revealed by the reduction in AVP and VIP mRNA levels demonstrated by in situ hybridization with radioactively labeled 48-mer AVP and 30-mer VIP probes. It is thus possible to conclude that the irreversible CET-induced changes in the neurochemistry of the SCN might underpin the disturbances in circadian rhythms observed after long-term alcohol consumption.

Key words: suprachiasmatic nucleus; chronic ethanol intake; withdrawal; AVP; VIP; GRP; somatostatin; stereology; immunocytochemistry; in situ hybridization

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