János Gardi

Brain corticotropin-releasing factor mediates ‘anxiety-like’ behavior induced by cocaine withdrawal in rats

Anxiety is a key symptom of the cocaine withdrawal syndrome in human addicts, and it is considered to be one of the major factors in precipitating relapse to chronic cocaine abuse. Corticotropin-releasing factor (CRF) plays an important role in the pathophysiology of anxiety and depression, and it may also be involved in the acute behavioral and neuroendocrine actions of cocaine. The role of endogenous CRF in cocaine withdrawal-induced anxiety was investigated in the present study. Animals were subjected to chronic cocaine (20 mg/kg, intraperitoneally, once a day for 14 days) administration. Rats tested 30 min after the last cocaine injection did not show withdrawal anxiety on the elevated plus maze or any alterations in brain CRF levels. Withdrawal (48 h) from chronic cocaine administration produced an intense anxiety-like behavior characterized by decreased open arm exploration. Immunoreactive CRF (CRF-LI) levels were selectively altered in the hypothalamus, in the amygdala and in the basal forebrain structures at the time of the behavioral anxiety, reflecting an increased activity of brain CRF systems. Daily intracerebroventricular (i.c.v.) pretreatment with an immunoserum raised against CRF completely prevented the development of anxiety induced by cocaine withdrawal. These data suggest that extrahypothalamic-limbic CRF hypersecretion may be involved in the development of anxiety related to cocaine withdrawal and that the CRF system may be a useful target for new pharmacotherapies for cocaine withdrawal and relapse.

Alterations of corticotropin-releasing factor-like immunoreactivity in different brain regions after acute cocaine administration in rats

Corticotropin-releasing factor (CRF) may mediate some of the neuroendocrine and behavioral responses to cocaine. In this study, the distribution of CRF-like immunoreactivity (CRF-LI) was determined in the hypothalamus and in several extrahypothalamic brain regions after acute cocaine administration in handled rats. CRF-LI decreased dose-dependently with cocaine administration in the hypothalamus and in the basal-forebrain structures. A small dose of cocaine (7.5 mg/kg) decreased CRF-LI in the hippocampus and in the frontal cortex. A significant, selective, dose-dependent increase in CRF-LI was found in the amygdala after cocaine injection. None of the investigated doses of cocaine altered CRF-LI in the striatum. These results suggest that acute cocaine administration alters brain CRF systems to contribute behavioral and neuroendocrine responses to cocaine.

GABAergic Neurogliaform Cells Represent Local Sources of Insulin in the Cerebral Cortex

Concentrations of insulin in the brain are severalfold higher than blood plasma levels. Insulin in the brain regulates the metabolism, molecular composition, and cognitive performance of microcircuits and reduces food intake; cerebral insulin levels are altered in diabetes, aging, obesity, and Alzheimers disease. Released by pancreatic β cells, insulin passes the blood–brain barrier, but sources of locally released insulin still remain unclear. We find that insulin is strongly expressed in GABAergic neurogliaform cells in the cerebral cortex of the rat detected by single-cell digital PCR. Focal application of glucose or glibenclamide to neurogliaform cells mimics the excitation suppressing effect of external insulin on local microcircuits via insulin receptors. Thus, neurogliaform cells might link GABAergic and insulinergic action in cortical microcircuits.

Calcitonin gene-related peptide levels in saliva of patients with burning mouth syndrome

Burning mouth syndrome (BMS) is an intraoral burning sensation for which no medical or dental cause can be found. Recent studies suggest that primary neuropathic dysfunction might be involved in the pathogenesis of BMS. Calcitonin gene-related peptide (CGRP) plays an important role in the development of pain and serves as a biological marker of trigeminovascular activation. The aim of this study was to determine the levels of CGRP in the saliva of BMS patients and estimate the trigeminovascular activation in BMS. CGRP levels were measured, by RIA method in 78 BMS patients and 16 healthy subjects. The levels of CGRP were non-significantly decreased in BMS patients in comparison to healthy subjects. These results suggest that trigeminal nerve degeneration may be the underlying cause of BMS.

Time-dependent alterations in corticotropin-releasing factor-like immunoreactivity in different brain regions after acute cocaine administration to rats

Recent data from various laboratories suggest that the activation of endogenous corticotropin-releasing factor (CRF) may contribute to the behavioral and neuroendocrine effects of cocaine. In the present study, the time-dependent variations in CRF-like immunoreactivity (CRF-LI) in the hypothalamus and several extrahypothalamic brain regions were determined after acute cocaine administration to handled rats. The intraperitoneal injection of 7.5 mg/kg cocaine led to a significantly decreased CRF-LI level in the basal forebrain and to a significantly increased CRF-LI level in the amygdala 60 min after administration, while the CRF-LI content was decreased in the hypothalamus and in the hippocampus 180 min after cocaine treatment. These results suggest that the durations of the effects of cocaine on CRF-LI are in the brain region-specific, which might contribute to the mediation of the diverse behavioral and neuroendocrine effects of cocaine.

INVOLVEMENT OF ENDOGENOUS CORTICOTROPIN-RELEASING FACTOR IN MEDIATION OF NEUROENDOCRINE AND BEHAVIORAL EFFECTS TO ALPHA-MELANOCYTE-STIMULATING HORMONE

The present work was to study if the α-melanocyte-stimulating hormone (α-MSH) was involved in activation of the pituitary-adrenal axis (PAA) in rats. The hormone increased plasma corticosterone (CORT) level, and induced an anxiogenic response as indicated by results from the elevated plus-maze test. Intracerebroventricular administration of corticotropin-releasing factor (CRF) antiserum (1:10, 1:20 and 1:100 dilutions in 1µl volume), overcame both the anxiogenic response and the PAA activating effect induced by α-MSH (50 µg s.c.) in a concentration-dependent manner. CRF antibody at the doses applied did not modify either the elevated plus-maze responses or CORT level by itself. Our results reveal that both the anxiogenic and the PAA activating effects of alpha-MSH are mediated by CRF.

Effects of testosterone on the rat renal medullary vasopressin receptor concentration and the antidiuretic response

The renal concentrating ability declines with age in humans and animals. Studies suggest that the concentrating defect is due to a decrease in renal vasopressin sensitivity. With ageing, expression of the renal vasopressin V2 receptor in rat is impaired; the normal receptor expression is restored by testosterone treatment. The effect of testosterone on the renal sensitivity to vasopressin was investigated in young rats. Male rats after orchidectomy and chronic antiandrogen cyproterone acetate treatment, and female rats after chronic testosterone phenylpropionate treatment, were used. The plasma arginine-vasopressin (AVP) and testosterone concentrations, and the antidiuretic responses to AVP and the V2 agonist deamino-[8-D-arginine]-vasopressin (dDAVP) after volume loading were measured, and the renal [3H]AVP binding density was determined. The plasma AVP level decreased slightly, but not significantly, in male rats after orchidectomy and cyproterone acetate treatment, but did not alter in female rats after testosterone treatment. The AVP and dDAVP sensitivities decreased in male rats after orchidectomy and cyproterone acetate administration, and increased in female rats treated with testosterone, as compared with the animals with a normal gonadal function. [3H]AVP binding to the renal inner medullary membranes was decreased following orchidectomy or antiandrogen treatment in male rats, and increased in testosterone-treated female rats. The results suggest that testosterone may play a physiological role in maintenance of the V2 vasopressin receptor expression and hence in the normal urinary concentrating ability in rat.

The somatostatin analog, octreotide, causes accumulation of growth hormone-releasing hormone and depletion of angiotensin in the rat hypothalamus

Rats were injected intracerebroventricularly with the somatostatin analog, octreotide (OCT; 0.1 microg) or vehicle, and hypothalamic contents of growth hormone-releasing hormone (GHRH), angiotensin II, and vasopressin were determined 10 min, 1, 3 and 6 h post-injection. OCT elicited an immediate release of angiotensin II (10 min) and a rise in GHRH content (1 h) followed by gradual (1-6 h) depletion of accumulated GHRH. Hypothalamic vasopressin was not altered but decreases in pituitary vasopressin occurred 10 min post-injection. The OCT-induced alterations in GHRH may explain previously reported changes in sleep whereas angiotensin may mediate OCT-induced drinking, vasopressin secretion and rises in blood pressure via sst2 somatostatin receptors.

The interaction of Urocortin II and Urocortin III with amygdalar and hypothalamic cotricotropin-releasing factor (CRF) – Reflections on the regulation of the hypothalamic–pituitary–adrenal (HPA) axis

Urocortin II (Ucn II) and Urocortin III (Ucn III) are selective agonists of the CRF receptor type 2 (CRFR2). The aim of the present experiments was to investigate the effects of Ucn II and Ucn III on the central CRF and peripheral glucocorticoids in rats. Increasing doses (0.5-1-2-5 μg/2 μl) of Ucn II or Ucn III were administered intracerebroventricularly, then CRF concentration was determined by immunoassays in two different brain regions, the amygdala and the hypothalamus, and in two different time paradigms, 5 and 30 min after the administration of peptides. In parallel with the second determination, plasma corticosterone concentration was measured by chemofluorescent assay. The amygdalar CRF amount was increased significantly by 0.5 and 5 μg of UCN II and 2 and 5 μg of UCN III in the 5 min experiments and by 5 μg of UCN II and 0.5 and 5 μg of UCN III in the 30 min experiments. The hypothalamic CRF content was not affected considerably in the 5 min paradigm, but it was influenced significantly in the 30 min paradigm, with 0.5 and 1 μg of UCN II and 0.5-2 μg of UCN III decreasing, and 2 and 5 μg of UCN II and 5 μg of UCN III increasing the hormone concentration, respectively. The plasma corticosterone concentration was decreased by 1 and 2 μg of UCN II and UCN III and increased by 0.5 and 5 μg of UCN III. The present results demonstrate that central administration of Ucn II and Ucn III modulate time-dependently and dose-dependently the amygdalar and the hypothalamic CRF concentration, and, directly or indirectly, the plasma corticosterone concentration. The present experiments suggest that the role of CRFR2 in the regulation of the HPA axis can be inhibitory or stimulatory, depending on the actual concentration of their agonists.

THE EFFECTS OF BRAIN AND C-TYPE NATRIURETIC PEPTIDES ON CORTICOTROPIN-RELEASING FACTOR IN BRAIN OF RATS

The central corticotropin-releasing factor (CRF)-ergic system plays a critical role in anxiety and other behavioral stress responses. It has been shown that atrial (ANP), brain (BNP) and C-type (CNP) natriuretic peptides exert anxiolytic-like effects in behavioral studies. Our previous findings demonstrated that various doses of centrally administered ANP selectively altered the CRF content in different brain areas. In the present study, CRF immunoreactivity was determined in hypothalamic and extrahypothalamic brain regions after central injection of BNP or CNP in rats. A high dose (400 ng) of BNP significantly increased the CRF-like immunoreactivity (CRF-LI) in the hypothalamus and amygdala, while only a tendency towards an increase was found in the hippocampus. In the hypothalamus, the CRF-LI decreased after a high dose (400 ng) of CNP. The CRF-LI increased in the basal forebrain after a low dose (100 ng) of CNP. These results suggest that CRF may be involved in the mediation of some neuroendocrine and behavioral responses to BNP and CNP.