Prolonged exposure to intermittent alcohol vapors blunts hypothalamic responsiveness to immune and non-immune signals.
BACKGROUND
We have previously shown that long-term alcohol treatment blunts the ACTH response to alcohol itself, as well as to other stresses, and is accompanied by decreased pituitary responsiveness to vasopressin (VP), but not corticotropin-releasing factor (CRF). The present work aims to determine the relevance of changes in CRF and VP receptors in the pituitary gland and/or peptide stores of CRF neurons in the paraventricular nucleus (PVN) of the hypothalamus, the areas that are most directly involved in ACTH release.


METHODS
Intact male rats were exposed to alcohol using a new vapor delivery system which enables individual rat housing in boxes. Alcohol treatment was delivered for 6 hr once daily (0700-1300), after which the rats were returned to their home cages where they had free access to food and water. Control rats were kept in similar boxes, but not exposed to alcohol. Total treatment time was 8 days. All animals were equipped with indwelling jugular cannulae that were used to monitor blood alcohol levels (BALs) as well as ACTH and corticosterone release throughout drug exposure. Due to the presence of a swivel, the animals' movements were not restricted or hindered by the presence of these cannulae. On the morning of day 9, the animals were decapitated under basal conditions or exposed to a neurogenic (mild electrofootshocks) or systemic [i.v. lipopolysaccharide (LPS)] stimulus. PVN neuronal responses, indicated by changes in mRNA concentrations of the immediate early genes (IEGs) c-fos and NGFI-B, and plasma ACTH levels were measured before and during endotoxemia or electrofootshocks.


RESULTS
In the absence of alcohol, plasma ACTH and corticosterone remained at basal levels, indicating the absence of environment-induced stress. In rats exposed to alcohol, BALs were consistent and predictable, and we targeted peak values of about 200 mg%. At the end of the drug treatment period, there were no significant differences between CRF and VP receptor mRNA levels in the anterior pituitary of control and alcohol-treated rats. In contrast, alcohol treatment respectively decreased CRF and increased VP stores in the external zone of the median eminence. It also increased NGFI-B and c-fos transcripts in the magnocellular (m) portion of the PVN, but not the parvicellular (p) division of this nucleus under basal conditions (i.e., in the absence of shocks or LPS). After exposure to these stressors, on the other hand, all groups of rats showed significant increases in plasma ACTH levels as well as up-regulation of their PVN neuronal response, as indicated by changes in pPVN IEGs transcripts. However, these hormonal and neuronal responses were significantly blunted in animals pretreated with alcohol.


CONCLUSIONS
Collectively, our results suggest that decreased PVN neuronal activation represents an important mechanism of the ability of long-term alcohol treatment to blunt the ACTH response to shocks or endotoxemia. In addition, the new system of alcohol delivery that we developed is practical and reliable, and has the significant advantage that it enables measurement of circulating hormone levels during drug exposure of the animals.
Lee
S
S
The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037, USA.
Schmidt
D
D
Tilders
F
F
Cole
M
M
Smith
A
A
Rivier
C
C
D000324:Adrenocorticotropic Hormone / Q000097:blood*; D000437:Alcoholism / Q000276:immunology*; D000818:Animals; D001143:Arousal / Q000187:drug effects* / Q000502:physiology; D019446:Endotoxemia / Q000276:immunology; D000431:Ethanol / Q000493:pharmacokinetics / Q000633:toxicity*; D007030:Hypothalamo-Hypophyseal System / Q000187:drug effects / Q000276:immunology; D007031:Hypothalamus / Q000187:drug effects* / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*; D008297:Male; D008856:Microscopy, Fluorescence; D010286:Paraventricular Hypothalamic Nucleus / Q000187:drug effects / Q000276:immunology; D010913:Pituitary-Adrenal System / Q000187:drug effects / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley
D008070:Lipopolysaccharides; D000431:Ethanol; D000324:Adrenocorticotropic Hormone