Rapid stress-induced elevations in corticotropin-releasing hormone mRNA in rat central amygdala nucleus and hypothalamic paraventricular nucleus: an in situ hybridization analysis

High densities of nerve cells containing corticotropin-releasing hormone (CRH) are located in the central nucleus of the amygdala (CeA) and paraventricular nucleus (PVN) of the hypothalamus. These brain regions play an important role in activating autonomic, behavioral, and endocrine responses to stress. This study was conducted to provide needed information concerning the acute effects of stress on CeA and PVN CRH mRNA expression. Rats were exposed to restraint stress for 1 h and brains collected after a 1-h post-stress interval. CRH mRNA expression occurring in the CeA and PVN was examined using in situ hybridization techniques. Densitometric analysis revealed that acute restraint stress produced significant increases in CRH mRNA levels in the PVN and in the rostral CeA region. In addition, the area in the rostral CeA encompassing high CRH mRNA signals increased significantly after stress. Results provide clear evidence that CRH neurons in the CeA and PVN exhibit rapid increases in CRH mRNA expression after exposure to stress.     

Serotonin stimulates corticotropin-releasing factor gene expression in the hypothalamic paraventricular nucleus of conscious rats

To examine the direct effects of serotonin (5-HT) on the release and synthesis of corticotropin-releasing factor (CRF) in the hypothalamic paraventricular nucleus (PVN), 5-HT was microinjected just onto the bilateral PVN of conscious rats. Plasma adrenocorticotropic hormone (ACTH) levels peaked at 30 min and returned to the basal levels in 90 min. Northern blot analysis revealed that the CRF messenger RNA (mRNA) level in the PVN as well as the proopiomelanocortin mRNA level in the anterior pituitary significantly increased 120 min after the 5-HT injections (50-250 nmol/side). Pretreatment with intracerebroventricular (i.c.v.) injection of pindobind 5-HT1A (5 nmol) or LY-278584 (500 nmol) completely abolished the 5-HT-induced ACTH response, whereas LY-53857 (100 nmol) was without effect. These results suggest that 5-HT stimulates CRF release, which has interactions with 5-HT1A and 5-HT3 receptors on CRF neurons in the PVN, and activates CRF synthesis in conscious rats.     

Reduced nicotinamide adenine dinucleotide phosphate-diaphorase activity in the paraventricular nucleus of the rat hypothalamus is modulated by estradiol

Leiomyomas represent 2% of gastric tumors. Commonly, gastric leiomyomas are clinically silent. Most often they become clinically apparent due to bleeding from ulceration of the overlying gastric mucosa. Surgical extirpation of the tumor is the standard treatment. Gastric leiomyomectomy was done routinely through open laparotomy until availability of laparoscopic equipment and techniques. Recently, there have been a few published reports regarding laparoscopic or laparoscopic-assisted removal of smooth muscle gastric tumors. There is little data, however, describing or discussing a laparoscopic approach to gastric leiomyomas located on the posterior gastric wall. We describe two different laparoscopic approaches to posterior wall gastric leiomyomas that we used in two patients. The postoperative recovery of both patients was remarkably quick and uneventful.     

Corticotropin releasing factor receptor type II (CRF2) messenger ribonucleic acid levels in the hypothalamic ventromedial nucleus of the infant rat are reduced by maternal deprivation

The stress neurohormone corticotropin releasing factor (CRF) activates at least two receptor types. Expression of corticotropin releasing factor receptor type II (CRF2) has been demonstrated in the hypothalamic ventromedial nucleus (VMH) of the adult and developing rat, but the physiological functions of VMH-CRF2 have not been elucidated. The VMH has been documented as an important participant in the regulation of food intake and its interactions with the hypothalamic-pituitary-adrenal axis and circadian rhythms. Regulation of VMH-CRF2 may thus play a role in the interplay of physiological alterations in metabolic state with the neuroendocrine and anorexic effects of CRF. This study determined the regulation of CRF2-mRNA expression in infant rats by the physiological consequences of maternal deprivation, i.e., fasting and stress. Using in situ hybridization, maternally deprived pups had an average 62% reduction of VMH-CRF2-mRNA levels compared with stress-free controls. Maternal deprivation also resulted in elevated plasma corticosterone levels (3.8 +/- 0.3 vs. 1.3 +/- 0.1 microg/dl) and an average 5.7% body weight loss. This study demonstrates that maternal deprivation, via fasting and HPA activation, leads to a dramatic decrease of CRF2-mRNA levels in the VMH. These results are consistent with a role for CRF2 activation in mediating some of the complex interactions of CRF (or urocortin) with regulation of food intake in the developing rat.     

Stimulation of neuropeptide Y overflow in the rat paraventricular nucleus by corticotropin-releasing factor

Neuropeptide Y (NPY) and corticotropin-releasing factor (CRF) are present at high concentrations in the hypothalamus where they mediate important endocrine and autonomic functions. Morphological and physiological studies have suggested an interaction between these peptides, and opposing actions of CRF and NPY have been reported on feeding and other behaviors. This study investigated the effect of CRF on NPY release in vivo, measured by push-pull techniques, in the anesthetized rat. Push-pull probes implanted into the paraventricular nucleus of the hypothalamus (PVN) were perfused with modified Ringer solution containing bovine serum albumin at 15 microl/min, and the perfusate was lyophilized prior to NPY radioimmunoassay. NPY overflow from the rat PVN was increased threefold by perfusion of a depolarizing concentration of potassium (50 mmol/L KCl). When CRF was administered into the PVN via the push-pull cannula at 1 or 5 microg/ml, dose-dependent increases in NPY overflow of two- and fivefold were observed (p < 0.05). These increases were abolished by prior intracerebroventricular (i.c.v.) administration of the CRF antagonist [D-Phe12,Nle(21,38),C(alpha)MeLeu32]CRF (12-41) at 1 or 5 microg/microl, respectively. NPY overflow returned promptly to resting levels following CRF administration. In contrast, when CRF was administered by i.c.v. bolus at a similar total dose (2 microg), no significant effect on NPY overflow was observed. These data provide in vivo evidence for an interaction between CRF and NPY at the level of the PVN.     

The arcuate nucleus is the major source for neuropeptide Y-innervation of thyrotropin-releasing hormone neurons in the hypothalamic paraventricular nucleus

Neuropeptide Y (NPY) immunoreactive (-ir) nerve fibers densely innervate hypophysiotropic TRH perikarya and dendrites in the hypothalamic paraventricular nucleus (PVN). To evaluate the contribution of the arcuate nucleus (Arc) to this innervation, the effect of Arc ablation by neonatal monosodium glutamate (MSG) treatment on the density of NPY-fibers contacting TRH neurons in the PVN was investigated. After the lesioned animals and vehicle-treated controls reached adulthood, the number of contacts between NPY-ir boutons and TRH-ir perikarya in the PVN was determined in double-immunostained sections. In controls, numerous contacts between NPY-ir terminals and TRH perikarya and dendrites were observed, confirming earlier findings. MSG treatment resulted in a marked reduction of the size of the Arc and also the number of NPY-perikarya with a concomitant reduction of 82.4 +/-2.1% in the relative number of NPY terminals contacting TRH perikarya and first order dendrites in the medial parvocellular and periventricular subdivisions of the PVN. In contrast, lesioning of the ascending adrenergic bundle in the brain stem caused no statistically significant change in the number of NPY-terminals in close apposition to hypophysiotropic TRH neurons in the PVN. These data confirm earlier findings that NPY-containing axon terminals innervate TRH neurons in the PVN and further demonstrate a potentially important anatomical relationship between NPY-producing neurons in the Arc and hypophysiotropic TRH neurons.     

Alterations in kappa opioid receptor mRNA levels in the paraventricular nucleus of the hypothalamus by stress and sex steroids

In this study, the levels of kappa opioid receptor (kOR) mRNA were determined using in situ hybridization following two types of stress (i.p. injection of hypertonic saline or novelty). In addition, we examined the possibility that estrogen or androgen would modify kOR mRNA. Gonadectomized male rats treated with estrogen or dihydrotestosterone were sacrificed 45 min after spending 15 min in a novel open field, or 60 min following hypertonic saline injection. Two-way ANOVA revealed that estrogen and novelty increased the levels of kOR mRNA in the ventral zone of the medial parvocellular part of the paraventricular nucleus (PVN), but not in the lateral parvocellular part of the PVN, claustrum, nucleus accumbens or the nucleus of the lateral olfactory tract. Furthermore, novelty increased kOR mRNA in gonadectomized (GDX) and GDX rats treated with dihydrotestosterone (DHTP), but not in sham-operated or estrogen-treated animals. Taken together, these data indicate that kOR mRNA levels are under estrogenic control and up-regulated in a stressor specific fashion.     

Depression in Parkinson's disease is not accompanied by more corticotropin-releasing hormone expressing neurons in the hypothalamic paraventricular nucleus

BACKGROUND: Depression is frequently encountered in Parkinson's disease (PD). In addition, more than half of the PD patients have a disturbed dexamethasone suppression test, which is associated with increased activity of corticotropin-releasing hormone (CRH) neurons. We recently found an increase in CRH neuron number, CRH-messenger RNA, and vasopressin colocalization in CRH neurons in the paraventricular nucleus (PVN) of depressed patients, which may be involved in the pathogenesis of depression. METHODS: The number of neurons expressing CRH was determined in the PVN of 6 depressed PD patients with a high score (> or = 13) on the Hamilton Depression Rating Scale, 6 nondepressed PD patients, and 6 controls. RESULTS: The three groups did not differ in the number of neurons expressing CRH. CONCLUSIONS: We hypothesize that activation of CRH neurons in the PVN, as we recently observed in idiopathic depression, does not play an essential role in depression in PD.     

Activation of neurons projecting to the paraventricular hypothalamic nucleus by intravenous lipopolysaccharide

The central nervous system interacts with the immune system to coordinate several components of the acute phase response, although the specific neuroanatomical pathways that mediate these responses are still uncharacterized. However, neurons in both the autonomic and endocrine components of the paraventricular hypothalamic nucleus (PVH) are characteristically activated in different models of immune stimulation. In the current study, we have used intravenous administration of lipopolysaccharide (LPS; 5 or 125 micrograms/kg) to induce the acute phase response. We subsequently coupled immunohistochemistry for Fos (as a marker of neuronal activation) with retrograde transport of the neuroanatomical tracer cholera toxin-b from the PVH. Several of the activated cell groups directly projected to the paraventricular nucleus, including the visceromotor (infralimbic) cortex, median preoptic nucleus, ventromedial preoptic area, bed nucleus of the stria terminalis, parabrachial nucleus, ventrolateral medulla, and nucleus of the solitary tract. These findings indicate that immune system stimulation activates cell groups from multiple nervous system levels that project to the paraventricular nucleus. We hypothesize that the activation of specific autonomic and endocrine elements of the PVH may be due to the activity of distinct afferents that converge on the PVH from multiple components of the central autonomic control system. Our results are consistent with the hypothesis that the PVH plays a key role in integrating diverse physiological cues into the varied manifestations that constitute the cerebral component of the acute phase response.     

The effects of fetal adrenalectomy at 110 days gestational age on AVP and CRH mRNA expression in the hypothalamic paraventricular nucleus of the ovine fetus

Previous studies with the granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) fusion protein, PIXY-321, demonstrated the enhanced biological activity of this molecule in comparison with GM-CSF or IL-3 alone or in combination. Here, we performed experiments to determine the proliferative effect of PIXY-321 on 13 constitutively growth factor-dependent human leukemia cell lines in comparison with GM-CSF, IL-3 and their combination using as read-out parameters the 48-hour 3H-thymidine incorporation assay and viable cell counts after in vitro culture for 7-8 days. Whereas one cell line was not responsive to any of these three cytokines, the other 12 cell lines showed variable degrees of growth in response to these effector molecules. PIXY-321 increased proliferation as measured by thymidine uptake relative to IL-3, GM-CSF or GM-CSF + IL-3 by 34% (range 5-448%), 12% (range 0-122%), and 6% (range 1-13%), respectively. PIXY-321 induced a mean increase of 32%, 30% and 11% in cell counts relative to the values obtained with IL-3, GM-CSF or GM-CSF + IL-3, respectively. Altogether, these data indicate that PIXY-321 stimulates proliferation of immature hematopoietic cells substantially better than equivalent concentrations of the single growth factors GM-CSF and IL-3. This hybrid growth factor showed a marginal to modest, but definite and reproducible increase in proliferation compared to the combination of GM-CSF plus IL-3. In summary, the fusion cytokine protein PIXY-321 appears to have biological effects superior to those elicited by its components, singly or in combination. This unique molecule should represent a useful tool in studies on the mechanisms underlying cytokine ligand-receptor interaction and the subsequent signal transduction. The use of PIXY-321 provides an opportunity for taking greater advantage in vitro and in vivo of the hematopoietic stimulatory activities of GM-CSF and IL-3.     

Electrophysiological activity in the central nucleus of the amygdala, emotionality and stress ulcers in rats.

Multiple-unit activity in the central nucleus of the amygdala was continuously recorded during 4 hr of restraint stress in rats. Five different activity profiles were found. Two types were associated with stress ulceration: one with increased stomach pathology, and the other with decreased stomach pathology. The same unit profiles were also differentially related to the emotionality characteristics of Wistar-derived rats, as well as to those of the genetically selected lines of Roman high- and low-avoidance rats. The type of profile that had been associated with increased pathology was generally seen in the Roman low-avoidance rats and in the Wistar rats that had been judged to be more emotional, that is, defecated before five "rearings" had occurred in an open-field test. The other unit profile was significantly more frequent in the Roman high-avoidance animals and the less emotional Wistar rats. Low-level electrical stimulation of both types of units produced stomach erosions in all cases. It was concluded that the unit activity in the central nucleus of the amygdala reflects certain emotionality characteristics of rats and also their susceptibility to stress ulcers. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Interleukin-1beta administered intracerebroventricularly stimulates the release of noradrenaline in the hypothalamic paraventricular nucleus via prostaglandin in the rat

The distribution of birth years for Nobel physicists shows a circaseptennian (about 7-year) periodicity. This observation extends an earlier observation of a circaseptennian pattern in the distribution of birth years for early quantum physicists. In both categories, birth rate tends to maximize in years belonging to the (7n + 4) phase.     

Neurons in the hypothalamic paraventricular nucleus send collaterals to the spinal cord and to the rostral ventrolateral medulla in the rat

A simple and sensitive HPLC method for determination of metronidazole in human plasma has been developed. A step of freezing the protein precipitate allowed an efficient separation of aqueous and organic phases minimizing the noise level and improved therefore the limit of quantitation (10 ng ml(-1) using 1 ml of plasma sample). The separation of compounds was performed on a RP 18 column with acetonitrile-aqueous 0.01 M phosphate solution (15:85, v/v) as mobile phase. Detection was performed by UV absorbance at 318 nm. Metronidazole was well resolved from the plasma constituents and internal standard. An excellent linearity was observed between peak-height ratios plasma concentrations over a concentration range of 0.01 to 10 microg ml(-1). Within-day and between-day precision (expressed by relative standard deviation) and accuracy (mean error in per cent) did not exceed 4% between 1 and 10 microg ml(-1) and 8.3 and 7.2% respectively for the limit of quantitation. The method is suitable for bioavailability and pharmacokinetic studies in humans.     

Levels of corticotropin-releasing hormone messenger ribonucleic acid (mRNA) in the hypothalamic paraventricular nucleus and proopiomelanocortin mRNA in the anterior pituitary during late gestation in fetal sheep

CRH regulates POMC gene expression and subsequent ACTH biosynthesis and release. In sheep, the preterm rise in fetal plasma ACTH commences at approximately 125 days gestation (dGA; 147 dGA = term), preceding the initiation of adrenocortical steroidogenesis. We hypothesized that an increase in CRH expression in the hypothalamic paraventricular nucleus (PVN) and POMC expression in the anterior pituitary in the late gestation sheep fetus may precede adrenal cortex maturation. Fetal sheep were obtained at 105-107 (n = 4), 128-130 (n = 5), and 138-140 (n = 4) dGA. Hypothalami were cryosectioned and subjected to in situ hybridization for ovine CRH mRNA. In all dGA groups, expression of CRH mRNA was observed throughout the rostrocaudal extent of the fetal PVN. The midrostral region of the fetal PVN where the dorsal and ventral divisions of the rostral PVN merge to form a single structure was selected for quantification. The number of copies of CRH probe hybridized per micron 3 were determined to estimate the quantity of hybridized CRH mRNA; the mean estimated CRH mRNA copy number per micron 3 midrostral PVN were 0.064 +/- 0.012 (105-107 dGA), 0.237 +/- 0.048 (128-130 dGA), and 0.108 +/- 0.034 (138-140 dGA; mean +/- SEM copies per micron 3 PVN). CRH mRNA signal significantly increased between 105-107 and 128-130 dGA (P < or = 0.05); 138-140 dGA levels of mRNA were not different from either 105-107 or 128-140 dGA levels. Regional variation in CRH mRNA levels were observed within the midrostral PVN between groups; at 138-140 dGA, a population of lateral midrostral PVN neurons maintain CRH mRNA levels greater than 105-107 dGA (P < 0.05), similar to those at 128-130 dGA. Fetal anterior pituitary RNA was subjected to Northern analysis for POMC mRNA. POMC mRNA levels in fetal anterior pituitaries were 14.1 +/- 2.2 (105-107 dGA), 28.9 +/- 10.9 (128-130 dGA), and 43.2 +/- 6 (138-140 dGA; mean +/- SEM arbitrary units). A significant increase (P < or = 0.05) was observed at 138-140 dGA compared to levels at 105-107 dGA. We conclude CRH mRNA levels in the fetal PVN increase coincident with increased POMC gene expression and the late gestation rise in fetal plasma ACTH. We speculate that a neuroendocrine stimulus at the fetal PVN may precipitate increased levels of CRH mRNA, initiating the maturation of the fetal hypothalamic-hypophyseal-adrenal axis, thus inducing the events of labor and delivery in sheep.     

RU-486 blocks stress-induced enhancement of proenkephalin gene expression in the paraventricular nucleus of rat hypothalamus

The purpose of this study was to investigate the glucocorticoid (GC) mediated regulation of corticotropin-releasing hormone (CRH) and proenkephalin (PE) gene expressions in the paraventricular nucleus (PVN) of the hypothalamus during physical stress induced by a single intraperitoneal (i.p.) injection of hypertonic saline (9% NaCl). Previous intracerebroventricular (i.c.v.) administration of the type II glucocorticoid receptor (GR) antagonist RU-486 (20 ng/rat), increased the basal CRH mRNA levels in the PVN but had no effect on PE gene expression. Stress induced by injection of hypertonic saline increased both CRH and PE mRNA levels in PVN. Administration of RU-486 completely blocked the stress-induced increase of PE mRNA levels, but failed to alter the CRH mRNA levels in the PNV. These data suggests that, under these experimental conditions, endogenous GC are necessary for a normal PE response to hypertonic saline stress.     

Effects of acidic fibroblast growth factor on neuronal activity of the parvocellular part in rat paraventricular nucleus

The effects of acidic fibroblast growth factor (aFGF) and its amino-terminal and carboxyl-terminal fragments (aFGF(1-15) and aFGF(114-140), respectively were examined on the neuronal activity in the parvocellular part of the paraventricular nucleus. As well known, this part contains a lot of corticotropin-releasing factor (CRF)-immunoreactive neurons. Application of 1 pg/ml and 2 pg/ml aFGF produced responses in 29.7% and 46.7% of neurons tested, respectively. Half or more than half of the responding neurons increased their discharge rate. Application of 0.2 ng/ml and 0.4 ng/ml aFGF(1-15) (1-15) also elicited response in 46.2% and 68.8% of neurons tested, respectively. Of these responding neurons, more than two third increased their firing rate. However, most of neurons tested for 0.67 ng/ml and 1.33 ng/ml aFGF(114-140) did not respond. Results suggest that aFGF and aFGF(1-15) promote the release of CRF through the activation of CRF-containing neurons.     

Neurons in the hypothalamic paraventricular nucleus that project to the rostral ventrolateral medulla are not activated by hypotension

The retrogradely-transported tracer, rhodamine-tagged microspheres was injected into the pressor region of the rostral ventrolateral medulla (RVLM) to enable detection of paraventricular neurons in the hypothalamus that project to the RVLM. The protein, Fos, was detected immunohistochemically and used to highlight neurons that were activated by hypotension (-16+/-5 mmHg) induced by diazoxide (30 mg/kg s.c.). Compared to controls, Fos production was increased by three-fold in the parvocellular paraventricular nucleus but there was no significant increase in the number of retrogradely-labelled cells that expressed Fos. The results suggest paraventricular nucleus (PVN) neurons projecting to the RVLM are not activated by hypotension.