Urocortin 1 administered into the hypothalamic supraoptic nucleus inhibits food intake in freely fed and food-deprived rats

Peptides of the corticotropin-releasing hormone/Urocortin (CRH/Ucn) family are known to suppress appetite primarily via CRH₂ receptors. In the rat hypothalamic supraoptic nucleus (SON), synthesis of both Ucn1 and CRH₂ receptors has been reported, yet little is known about the effects of Ucn1 in the SON on feeding behaviour. We first established the dose-related effects of Ucn1 injected into the SON on the feeding response in both freely fed and 24-h food-deprived rats. A conditioned taste avoidance paradigm was performed to investigate possible generalised effects of local Ucn1 treatment. Administration of Ucn1 into the SON at doses equal to or higher than 0.5 μg significantly decreased food intake in both freely fed and food-deprived rats. The Ucn1-mediated suppression of food intake was delayed in freely fed as compared to food-deprived animals. Conditioning for taste aversion to saccharine appeared at 0.5 and 1 μg of Ucn1. Both the early and the delayed onset of anorexia observed after intra-SON injection of Ucn1 under fasting and fed conditions, respectively, suggest the possible involvement of different CRH receptor subtypes in the two conditions, while the conditioned taste aversion seems to be responsible for the initial latency to eat the first meal in these animals.     

Inhibitory response in neurosecretory cells of the hypothalamic supraoptic nucleus to subiculum stimulation in lactating rats

The effects of subiculum stimulation were investigated in 80 antidromically identified hypothalamic supraoptic neurons in lactating rats. Inhibition manifesting as suppression of antidromic action potentials (or of their somatodendritic component) was revealed in 26% of cells, induced by applying conditioned and test stimuli to the subiculum and neurohypophysial stalk. In some instances inhibition arose following a latency of 5–25 msec after each subicular stimulus and lasted only briefly; it set in gradually in other cases, leading to stable long-term changes in the excitability of neurosecretory cells. No activation was produced by this stimulation. It is deduced that subicular inhibitory inputs follow different patterns, thus reflecting morphological organizational aspects of synaptic inhibitory inputs to neurosecretory cells.A. A. Ukhtomskii, Institute of Physiology, A. A. Zhdanov State University, Leningrad. Translated from Neirofiziologiya, Vol. 20, No. 4, pp. 431–437, July–August, 1988.     

Investigating the role of the hypothalamic supraoptic nucleus in nociception in the rat

We investigated the role of the hypothalamic supraoptic nucleus (SON) in nociception in the rat. Electrical stimulation of the SON or microinjection of a small dose of L-glutamate sodium into the SON elevated the nociceptive threshold in a dose-dependent manner, while cauterization of the SON decreased the nociceptive threshold. Pituitary removal did not influence the antinociceptive effect of L-glutamate sodium in the SON. The data suggested that the neurons and not the nervous fibers in the SON played an important role in antinociception.     

Ultrastructure of neurosecretory cells of the hypothalamic supraoptic nucleus in white rats following painful stress and during aging

The ultrastructural organization of neurosecretory cells (NSC) belonging to the hypothalamic supraoptic nucleus was investigated in young rats following periods of painful stress differing in duration (of 2 and 20 min). Intact young rats of similar age and others at late stages of development (24–27 months old) served as controls. Short- and longer-acting painful stress was found to intensify and inhibit the functional activity of NSC respectively. Complex ultrastructural changes in the NSC of young rats following protracted painful stress is compared with the ultrastructural organizational pattern in several NSC of aging rats. Findings would imply that aging occurs in rats following prolonged painful stress, resulting from functional hypersecretion and depletion.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 3, pp. 389–395, May–June, 1989.     

Neurosecretory activity in supraoptic nucleus of normal rats

Summary The secretory rhythm of neurosecretory cells was studied in supraoptic nuclei of normally drinking rats. Both nuclei were divided in five zones, and in each one, the nuclear and nucleolar volumes of 50 neurons were measured. Parallel observations were made on the Nissl material and neurosecretory granules. Comparative effects of different fixatives showed Carnoy and Zenker to be the best fluids for this type of investigation.Two extreme types of neurosecretory neurons with intermediary stages are observed. Type I neuron has a small nucleus and nucleolus, well developed Nissl bodies and a greater content of neurosecretory material. Type II has larger nucleus and nucleolus, scanty or packed Nissl bodies, and little Gomori positive material. The nuclear and nucleolar volumes differ between the different zones of the supraoptic nucleus and the zone of maximal activity varies in each particular case. To explain this asynchronic behavior a neurosecretory wave of activity moving along the supraoptic nucleus is postulated.Supported by Grants of the Consejo Nacional de Investigaciones Científicas y Técnicas, and the Air Force Office of Scientific Research 314-64.Fellows of the Consejo Nacional de Investigaciones Cientificas y Técnicas.     

Different pharmacological anatomy in the paraventricular hypothalamic nucleus, supraoptic nucleus, and suprachiasmatic nucleus of rats: quantitative autoradiography on angiotensin II receptor binding sites

Angiotensin II (AII) and vasopressin (VP) play important roles in cardiovascular function. Using 125I-[Sar1,Ile8]-angiotensin II (125I-SI-AII), a potent AII antagonist, AII receptor binding sites were autoradiographically localized in three VP-producing areas of the hypothalamus and compared in hypertensive and normotensive rats. Within three major VP-producing areas, AII receptor binding was highest in the paraventricular hypothalamic nucleus and lowest in the supraoptic nucleus, suggesting that a differential AII regulation of separate VP systems exists in the brainstem. No statistical difference in 125I-SI-AII receptor binding was found between WKY and SHR rats in each of the three major VP-producing nuclei studied. These results are consistent with a role of AII receptors in a subtle and complicated regulation of VP in cardiovascular function.     

Modifications of neuronal activity in the rat hypothalamic supraoptic nucleus after long-lasting vibrational stimulation

We analyzed the background impulse activity (BIA) generated by neurons of the rat hypothalamic supraoptic nucleus in the norm and under conditions of long-lasting vibrational stimulation (exposure 5, 10, or 15 days). Distributions of neurons by the level of regularity, dynamics of discharge trains, form of histograms of interspike intervals (ISIs), as well as distributions of neurons by the BIA frequency ranges, were studied. We also calculated the mean frequency of impulsation of the neurons under study and the coefficient of variation of ISIs. After vibrational influences, we found modifications of both the internal structure of the recorded spike trains and the mean frequency of impulsation within the entire studied group and different frequency subgroups. Neirofiziologiya/Neurophysiology, Vol. 38, No. 3, pp. 224–230, May–June, 2006.     

Background activity of neurons of the supraoptic hypothalamic nucleus in rats under conditions of vibrational stimulation and electromagnetic extrahigh-frequency irradiation

We analyzed background impulse activity of neurons of the supraoptic nucleus of the rat hypothalamus in the course of 15-day-long isolated action of generalized vibrational stimulation and combination of such stimulation with irradiation of the animal’s head with low-intensity extrahigh-frequency (EHF, millimeter-range) electromagmetic waves. The distributions of the neurons by the level of regularity and dynamics of spike trains, separate frequency ranges of impulsation, and pattern of interspike interval (ISI) histograms were estimated. We also calculated the mean frequency of discharges and coefficient of variation of ISIs. A trend toward decreases in the deviations of some parameters of neuronal spike activity generated by supraoptic neurons, which were evident within early time intervals of isolated action of vibration (5 to 10 days), was observed under the influence of EHF electromagnetic irradiation; thus, the latter factor probably exerts a sedative effect. Neirofiziologiya/Neurophysiology, Vol. 39, No. 6, pp. 433–442, November–December, 2007.     

Injection of noradrenaline into the hypothalamic paraventricular nucleus produces vigorous gnawing in satiated rats

Single injections of noradrenaline (NA) into the hypothalamic paraventricular nucleus (PVN) initiate short bouts of eating in the rat. The effect of the injections of NA (40 nM) into the PVN on gnawing was studied in satiated male rats that had displayed a reliable eating response to NA. The NA injections produced numerous bouts of prolonged and vigorous gnawing of wood pieces shaped like food pellets. The gnawing response started 0.5-2 min after the injection of NA and lasted for 20-25 min. The results suggest that the increase in gnawing and eating evoked by NA might be related to facilitation of the oral responses, and a changed sensitivity to the food related stimuli, in addition to an intrinsic increase in appetite.     

Reciprocal pathway between medullary visceral zone and hypothalamic supraoptic nucleus or paraventricular nucleus involved in hyperosmotic regulation

In this study we try to simultaneously investigate the response of neurons and astrocytes of rats following hyperosmotic stimulation and test the possibility that the reciprocal pathways between medullary visceral zone (MVZ) and hypothalamic paraventricular nucleus (PVN) or supraoptic nucleus (SON). Hyperosmotic pressure animal model was established by administering 3% sodium chloride as drinking water to rats. The distribution and expression of the HRP retrogradely labeled neurons, Fos, tyrosine hydroxylase (TH) or vasopressin (VP) positive neuron and glial fibrillary acidic protein (GFAP) positive astrocytes in the MVZ, SON and PVN were observed by quadruplicate-labeling methods of WGA-HRP retrograde tracing combined with anti-Fos, TH (or VP) and GFAP immunohistochemical technique. Fos positive neurons within the MVZ, PVN and SON increased markedly. There were also a large number of GFAP positive structures in the brain and their distribution pattern was fundamentally similar or analogous to Fos positive neurons in the above-mentioned areas. The augmented GFAP reactivities took on hypertrophic cell bodies, thicker and longer processes. Quadruplicate immunohistochemical staining showed that a neuron could be closely surrounded by many astrocytes and they formed neuron-astrocytic complex (N-ASC). Fos+/TH+/HRP+/GFAP+ and Fos+/VP+/HRP+/GFAP+ quadruplicate labeled N-ASC could be found in the MVZ, PVN and SON, respectively. The present results indicated that the neurons and astrocytes might be very active following hyperosmotic pressure and N-ASC as a functional unit might serve to modulate osmotic pressure. There were reciprocal osmoregulation pathways between the MVZ and SON or PVN in the brain.     

Orexin A (hypocretin 1) injected into hypothalamic paraventricular nucleus and spontaneous physical activity in rats

In humans, nonexercise activity thermogenesis (NEAT) increases with positive energy balance. The mediator of the interaction between positive energy balance and physical activity is unknown. In this study, we address the hypothesis that orexin A acts in the hypothalamic paraventricular nucleus (PVN) to increase nonfeeding-associated physical activity. PVN-cannulated rats were injected with either orexin A or vehicle during the light and dark cycle. Spontaneous physical activity (SPA) was measured using arrays of infrared activity sensors and night vision videotaped recording (VTR). O(2) consumption and CO(2) production were measured by indirect calorimetry. Feeding behavior was assessed by VTR. Regardless of the time point of injection, orexin A (1 nmol) was associated with dramatic increases in SPA for 2 h after injection (orexin A: 6.27 +/- 1.95 x 10(3) beam break count, n = 24; vehicle: 1.85 +/- 1.13 x 10(3), n = 38). This increase in SPA was accompanied by compatible increase in O(2) consumption. Duration of feeding was increased only when orexin A was injected in the early light phase and accounted for only 3.5 +/- 2.5% of the increased physical activity. In a dose-response experiment, increases in SPA were correlated with dose of orexin A linearly up to 2 nmol. PVN injections of orexin receptor antagonist SB-334867 were associated with decreases in SPA and attenuated the effects of PVN-injected orexin A. Thus orexin A can act in PVN to increase nonfeeding-associated physical activity, suggesting that this neuropeptide might be a mediator of NEAT.     

In vitro culture of the hypothalamic supraoptic nucleus. I. General morphological characteristics

Kinetics and morphology of supraoptic nuclei (SON) organ culture of newborn rat hypothalamus have been described. Explants of SON were cultured for 7-90 days. Cell migration, growth of neuron sprouts, formation of nerve bundles and growth zone, and mitotic activity were followed. Neurosecretory cells (NSC) differing in size, shape and degree of activity, were identified in addition to other cell elements of SON. The neurosecret was shown to appear at the beginning of the second week of cultivation in large and middle size NSC. In small and old cultures of NSC no neurosecrete was discovered. It was established that NSC complete the terminal differentiation and maintain for a long time their viable and functional activity in vitro.     

Mechanism of the cardiovascular responses caused by l-proline microinjected into the supraoptic nucleus of the hypothalamus in unanesthetized rats

In the present study, we report on the cardiovascular effects caused by the microinjection of l-proline (l-Pro) into the supraoptic nucleus (SON) in unanesthetized rats: the possible involvement of ionotropic glutamate receptors in the SON, as well as the peripheral mechanisms involved in the mediation of its cardiovascular effects. We compared the l-Pro effects with those caused by the injection of l-glutamate (l-Glu) into the SON. Microinjection of increasing doses of l-Pro into the SON caused dose-related cardiovascular responses in unanesthetized rats that were similar to those observed after the injection of l-Glu. Pretreatment of the SON with either a selective non-NMDA (NBQX) or a selective NMDA (LY235959) glutamate receptor antagonist blocked the cardiovascular response to l-Pro. The dose–effect curve for the pretreatment with increasing doses of LY235959 was shifted to the left in relation to the curve for NBQX, showing that LY235959 is more potent than NBQX in inhibiting the cardiovascular response to l-Pro. On the other hand, the cardiovascular response to l-Glu was only significantly reduced by pretreatment with NBQX (2 nmol/100 nL), but not affected by LY235959 (2 nmol/100 nL). The pressor response to l-Pro was not affected by intravenous pretreatment with the ganglion blocker pentolinium, but it was blocked by intravenous pretreatment with the V₁-vasopressin receptor antagonist dTyr(CH₂)₅(Me)AVP. In conclusion, these results suggest that l-Pro has a selective receptor that is sensitive to ionotropic glutamate receptor antagonists. Its activation in the SON results in vasopressin release into the systemic circulation, causing pressor and bradycardiac responses.     

Oxytocin, but not arginine vasopressin is involving in the antinociceptive role of hypothalamic supraoptic nucleus

Our pervious study has demonstrated that the hypothalamic supraoptic nucleus (SON) plays a role in pain modulation. Oxytocin (OXT) and arginine vasopressin (AVP) are the important hormones synthesized and secreted by the SON. The experiment was designed to investigate which hormone was relating with the antinociceptive role of the SON in the rat. The results showed that (1) microinjection of l-glutamate sodium into the SON increased OXT and AVP concentrations in the SON perfusion liquid, (2) pain stimulation induces OXT, but not AVP release in the SON, and (3) intraventricular injection (pre-treatment) with OXT antiserum could inhibit the pain threshold increase induced by SON injection of l-glutamate sodium, but administration of AVP antiserum did not influence the antinociceptive role of SON stimulation. The data suggested that the antinociceptive role of the SON relates to OXT rather than AVP.     

Anxiolytic effects of lavender oil inhalation on open-field behaviour in rats

To establish a valid animal model of the effects of olfactory stimuli on anxiety, a series of experiments was conducted using rats in an open-field test. Throughout, effects of lavender oil were compared with the effects of chlordiazepoxide (CDP), as a reference anxiolytic with well-known effects on open-field behaviour. Rats were exposed to lavender oil (0.1-1.0 ml) for 30 min (Experiment 1) or 1h (Experiment 2) prior to open-field test and in the open field or injected with CDP (10 mg/kg i.p.). CDP had predicted effects on behaviour, and the higher doses of lavender oil had some effects on behaviour similar to those of CDP. In Experiment 3, various combinations of pre-exposure times and amounts of lavender oil were used. With sufficient exposure time and quantity of lavender the same effects were obtained as in Experiment 2. Experiment 4 demonstrated that these behavioural effects of lavender could be obtained following pre-exposure, even if no oil was present in the open-field test. In Experiments 2-4, lavender oil increased immobility. Together, these experiments suggest that lavender oil does have anxiolytic effects in the open field, but that a sedative effect can also occur at the highest doses.     

Topochemical aspects of nucleic-acid and protein metabolism within the neuron-neuroglia unit of the hypothalamic supraoptic nucleus

The RNA and both the total and basic protein content of individual cells were determined by cytospectrophotometry in neurons and perineuronal oligodendroglia of the hypothalamic supraoptic nucleus in rats subjected to various stresses, as well as in ground squirrels during natural hibernation. Barbiturate narcosis and deep cooling, which induced a decrease in body temperature in rats and hibernation in squirrels, caused a marked decrease of all macromolecular constituents in neurons. A similar decrease was found in the perineuronal oligodendroglia in rats, but an increase was observed in ground squirrels. After cessation of cooling, while the body temperature of the animals returned to normal, the neurons, but not the oligodendroglia, of rats showed a significant accumulation of RNA, while RNA accumulated in both neurons and perineuronal oligodendroglia in ground squirrels. Milder cooling of rats, which did not lower their body temperature, induced reciprocal changes in basic-protein content in neuronal and glial cell nuclei, with the accumulation of protein occurring initially in neurons, and subsequently in glia. When cold adaptation was accomplished, the basic protein content of neurons and glial cells returned to the control level. Four days after adrenalectomy in rats, the RNA content decreased in oligodendroglia but not in neurons of the supraoptic nucleus. This effect was completely abolished by daily injections of cortisol in the adrenalectomized animals. The data obtained indicate the existence of differences in metabolic responses to stress between neurons and glial cells of the supraoptic nucleus of the hypothalamus.     

Neuropeptidomics of the supraoptic rat nucleus

The mammalian supraoptic nucleus (SON) is a neuroendocrine center in the brain regulating a variety of physiological functions. Within the SON, peptidergic magnocellular neurons that project to the neurohypophysis (posterior pituitary) are involved in controlling osmotic balance, lactation, and parturition, partly through secretion of signaling peptides such as oxytocin and vasopressin into the blood. An improved understanding of SON activity and function requires identification and characterization of the peptides used by the SON. Here, small-volume sample preparation approaches are optimized for neuropeptidomic studies of isolated SON samples ranging from entire nuclei down to single magnocellular neurons. Unlike most previous mammalian peptidome studies, tissues are not immediately heated or microwaved. SON samples are obtained from ex vivo brain slice preparations via tissue punch and the samples processed through sequential steps of peptide extraction. Analyses of the samples via liquid chromatography mass spectrometry and tandem mass spectrometry result in the identification of 85 peptides, including 20 unique peptides from known prohormones. As the sample size is further reduced, the depth of peptide coverage decreases; however, even from individually isolated magnocellular neuroendocrine cells, vasopressin and several other peptides are detected.     

Pattern of response in neurosecretory cells of the rat hypothalamic supraoptic nucleus to subiculum stimulation under water deprivation

The effects of water deprivation were investigated in the pattern of response produced by subiculum stimulation in antidromically identified hypothalamic supraoptic neurosecretory cells of lactating rats. In dehydrated animals as compared with the controls, the percentage of neurons responding to subiculum stimulation with an inhibitory action (blockade of antidromic action potential) remained unchanged, although the proportion of differing inhibitory response did alter: numbers of cells with gradually developing inhibitory response increased significantly and fewer cells showed transitory development of inhibition. Inhibitory response emerging as depression of background spike activity showed a quantitative increase, moreover. Plasticity was found to be one distinguishing feature of afferent input from the subiculum to supraoptic nucleus neurosecretory cells and, in particular, a capacity for reorganization under water deprivation.A. A. Ukhtomskii Institute of Physiology, State University, Leningrad. Translated from Neirofiziologiya, Vol. 22, No. 2, pp. 243–249, March–April, 1990.     

经脂多糖（LPS）处理免疫应答大鼠离体脑片视上核神经元对细胞因？…

实验采用离体脑片全细胞膜片箝记录方法,观察了细胞因子白介素－１β（ＩＬ－１β）和ＩＬ－２对大鼠离体脑片视上核神经元膜电位及自发放电的影响,以期探明免疫应答大鼠视上核神经元对细胞因子敏感性的变化。结果显示,用１００Ｕ／ｍｌ ＩＬ－１β藻流脑片,正常对照的（ｎ＝１５）和脂多糖（ｌｉｐｏｐｏｌｙｓａｃｃｈｙａｒｉｄｅ ＬＰＳ）腹腔注射９ｄ的大鼠视上核神经元（ｎ＝２０）超极化,同时伴有自发放电频率的下降;