Plasticity of neuroendocrine transducers under the combined influence of the radiation and light exposure

The structural changes of neurons of the rat hypothalamic supraoptic (SON) and of paraventricular (PVN) nucleus after 48 h of bright light exposure, of 5 Gy whole-body X-irradiation and of their combination subjected to the analysis by means of light-optic and of electron microscopy for the estimation of radimodificated effect of light exposure lasted 24 h a day and plasticity of neuroendocrine transducers interacted with the optic sensory system. The structural changes of neurons of the SON after combined action are less considerable and more prolonged in comparison with the PVN that loas defermined by their direct connection with the optic sensory system via the retinohypothalamic tract.     

Aromatase immunoreactivity in the rat brain: Gonadectomy-sensitive hypothalamic neurons and an unresponsive “limbic ring” of the lateral septum-bed nucleus-amygdala complex

The aromatase (estrogen synthetase) enzyme catalyzes the conversion of androgens to estrogens in peripheral tissues, as well as in the brain. Our study aimed at comparing the brain distribution of aromatase-immunoreactive neurons in male and female, normal and gonadectomized rats. Light microscopic immunostaining was employed using a purified polyclonal antiserum raised against human placental aromatase. Two anatomically separate aromatase-immunoreactive neuronal systems were detected in the rat brain: A “limbic telencephalic” aromatase system was composed by a large population of labeled neurons in the lateral septal area, and by a continuous “ring” of neurons of the laterodorsal division of the bed nucleus of stria terminalis, central amygdaloid nucleus, stria terminalis, and the substantia inominata-ventral pallidum-fundus striati region. The other, “hypothalamic” aromatase system consisted of neurons scattered in a dorsolateral hypothalamic area including the paraventricular, lateral and dorsomedial hypothalamic nuclei, the subincertal nucleus as well as the zona incerta. In addition, a few axon-like processes (unresponsive to gonadectomy) were present in the preoptic-anterior hypothalamic complex, the ventral striatum, and midline thalamic regions. No sexual dimorphism was observed in the distribution or intensity of aromatase-immunostaining. However, 3 days, 2, 3, 8, 16, or 32 weeks after gonadectomy, aromatase-immunoreactive neurons disappeared from the hypothalamus, whereas they were still present in the limbic areas of both sexes. The results indicate the existence of two distinct estrogen-producing neuron systems in the rat brain: (1) a “limbic ring” of aromatase-labeled neurons of the lateral septum-bed nucleus-amygdala complex unresponsive to gonadectomy; and (2) a sex hormone-sensitive “hypothalamic” aromatase neuron system.     

氨甲酰胆碱引起的促钠排泄与下丘脑TH—IR神经元活性的变化

目的：我们最近的实验发现大鼠侧脑室注射氨甲酰胆碱引起显著的促钠排泄作用,本工作同时还观察了下丘脑内不同脑区的儿茶酚胺能神经元活性的变化。方法和结果：氨甲酰胆碱注射后４０ｍｉｎ,下丘脑室旁核的腹侧和内侧小细胞部、内侧视前区、尾核、苍白球的酪氨酸羟化酶免疫反应（ｔｈｙｒｏｓｉｎｅｈｙｄｒｏｘｙｌａｓｅｉｍｍｕｎｏｒｅａｃｔｉｖｉｔｙ,ＴＨＩＲ）阳性细胞数减少,免疫反应染色强度降低;下丘脑室旁核的后部,下丘脑前区的后部、下丘脑室周核、弓状核、下丘脑外侧区的ＴＨＩＲ阳性细胞数增多,免疫反应染色强度增强。结论：侧脑室注射氨甲酰胆碱对脑内不同脑区的内源性儿茶酚胺能神经元分别有兴奋或抑制作用,其与促钠排泄的关系将在本文中讨论     

褪黑素对大鼠下丘脑薄片视交叉上核神经元放电昼夜节律性的调制

先用持续光照和松果腺切除预处理大鼠,然后制成下丘脑薄片,记录其视交叉上核（ＳＣＮ）神经元自发放电,观察其昼夜变化和褪黑素（ＭＥＬ）对它的影响。实验结果表明：⑴在正常光照（光照：黑暗＝１２：１２）条件下,ＳＣＮ神经元自发放电频率呈现昼夜低的节律性。在昼夜时间（ＣＴ）６－８出现放电高峰,频率约为８．３Ｈｚ;在ＣＴ１８－２０出现低谷,频率约为３．８Ｈｚ。松果腺切除后,ＳＣＮ神经元自发放电的昼夜节律性基本     

Immunocytochemical localization of somatostatin-containing neurons in the rat hypothalamus

Summary The rat hypothalamus was studied at the light microscopic level with the use of single and double immunocytochemical staining methods. It was shown that the rat supraoptic and paraventricular hypothalamic nuclei, and their accessory neurosecretory nuclei, do not contain magnocellular somatostatin neurons. The distribution of the hypothalamic parvocellular somatostatin cells is described. The parvocellular component of the rat hypothalamic paraventricular nucleus is, at least partly, composed of somatostatin cells: they form a fairly well circumscribed periventricular cell mass. The rat suprachiasmatic nuclei contain separate somatostatin neurons and vasopressin neurons. Scattered somatostatin cells are present in the entire arcuate nucleus. In addition to the periventricular somatostatin cells located in the preopticanterior hypothalamic area and in the arcuate nucleus, the rat hypothalamus also contains numerous scattered somatostatin cells located distant from the third ventricle.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

兔脑内Orexin B免疫阳性神经元的分布定位

采用免疫组织化学方法研究了10只青紫蓝兔脑内Orexin B免疫阳性神经元的分布定位。结果显示,Orexin B免疫阳性神经元分布于下丘脑的室旁核、背内侧核、穹隆周核、外侧区和后区以及底丘脑的未定带。以下丘脑背内侧核、穹隆周核和外侧区的阳性神经元数量较多,下丘脑室旁核、后区和未定带较少。表明了兔脑内Orexin B免疫阳性神经元的分布与Orexin A免疫阳性神经元的分布存在一些差异,提示两种Orexin的产生部位和生理功能可能也存在差异。     

大鼠下丘脑内一氧化氮合酶阳性神经元的分布

用ＮＡＤＰＨ－ｄ组织化学方法观察了大白鼠下丘脑内一氧化氮合酶（ＮＤＳ）阳性神经元的分布及形态特征。结果显示：在视上核、室旁核的大细胞部、环状核、穹窿周核、下丘脑外侧区、下丘脑腹内侧核、下丘脑背内侧核、乳头体区大部分核团均可见一氧化氮合酶阳性神经元聚集成团。在视前内侧区、视前外侧区、下丘脑前区、下丘脑背侧区、下丘脑后区、室周核、室旁核小细胞部及穹窿内可见散在的一氧化氮合酶阳性神经元。室周核内可见呈阳性反应的接触脑脊液神经元的胞体及突起。一氧化氮合酶阳性神经元大多可见突起,有的突起上可见１～２级分支,并可见膨体。下丘脑大部分区域内可见阳性神经纤维。弓状核内可见许多弧形纤维连于第三脑室室管膜和正中隆起。     

Ultrastructural features of the lateral preoptic area,median eminence and arcuate nucleus of the rat

Summary The arcuate nucleus, median eminence, and the lateral preoptic area from the brains of aldehyde-perfused male and female rats were examined by electron microscopy. In the lateral preoptic area, three neuronal types are described: a small light neuron, a larger light one, and a dark neuron resembling the larger light one in size and nuclear shape. Many myelinated axons are interposed among single neurons or neuronal pairs. The relationship of structures to each other is discussed. Several observations not previously reported are illustrated from tissue of the arcuate nucleus and median eminence.     

Hypothalamic galanin is up-regulated during hyperphagia and increased body weight gain induced by disruption of signaling in the ventromedial nucleus

Disruption of signaling in the ventromedial nucleus (VMN) by colchicine (COL) produces transient (4 days) hyperphagia and weight gain. Microinjection of galanin into various hypothalamic sites stimulates feeding, so we tested the hypothesis that galanin is up-regulated in COL-treated rats by analyzing galanin concentrations in micropunched hypothalamic sites. Galanin was increased in the paraventricular nucleus on Days 1 through 4 after COL-injection. Galanin was also elevated in three other hypothalamic sites, the dorsomedial nucleus, lateral hypothalamic area, and perifornical hypothalamus, on Days 2-4 and in the lateral preoptic area, on Day 1 only. In the median eminence-arcuate nucleus and amygdala an initial decrease on Day 1 was followed by a then progressive increase through Day 4. These increases occurred despite marked elevations in blood insulin and leptin, hormones known to suppress hypothalamic galanin. When COL- or saline-treated rats were injected intracerebroventricularly with galanin, it stimulated feeding further in the hyperphagic COL-treated rats, but the relative response over basal consumption was similar in both COL-treated and control rats. These results in VMN disrupted rats suggest that neurochemical rearrangements, including increased availability of galanin, may contribute to the hyperphagia and increased weight gain; additionally, it seems that neurons in the VMN normally exert a restraint on galanin signaling.     

Coexistence of substance P and neurotensin-like peptides in single neurons of the rat hypothalamus

The coexistence of substance P with neurotensin-like immunoreactivity in certain neurons of the hypothalamus were demonstrated by the double immunofluorescence method. Substance P and neurotensin-like immunoreactivity coexisted within single neurons of some hypothalamic areas such as the medial preoptic area, perifornical area, anterior hypothalamic area, lateral hypothalamic area, periventricular nucleus and posterior hypothalamic nucleus, although they did not coexist in the majority of immunoreactive cells.     

Prenatal exposure to ethanol causes partial diabetes insipidus in adult rats

Chronic consumption of ethanol in adult rats and humans leads to reduced AVP-producing neurons, and prenatal ethanol (PE) exposure has been reported to cause changes in the morphology of AVP-producing cells in the suprachiasmatic nucleus of young rats. The present studies further characterize the effects of PE exposure on AVP in the young adult rat, its hypothalamic synthesis, pituitary storage, and osmotically stimulated release. Pregnant rats were fed a liquid diet with 35% of the calories from ethanol or a control liquid diet for days 7-22 of pregnancy. Water consumption and urine excretion rate were measured in the offspring at 60-68 days of age. Subsequently, the offspring were infused with 5% NaCl at 0.05 ml.kg(-1).min(-1) with plasma samples taken before and at three 40-min intervals during infusion for measurement of AVP and osmolality. Urine output and water intake were approximately 20% greater in PE-exposed rats than in rats with no PE exposure, and female rats had a greater water intake than males. The relationship between plasma osmolality and AVP in PE-exposed rats was parallel to, but shifted to the right of, the control rats, indicating an increase in osmotic threshold for AVP release. Pituitary AVP was reduced by 13% and hypothalamic AVP mRNA content was reduced by 35% in PE-exposed rats. Our data suggest that PE exposure can cause a permanent condition of a mild partial central diabetes insipidus.     

Hormone secretion in transgenic rats and electrophysiological activity in their gonadotropin releasing-hormone neurons

Expression of GFP in GnRH neurons has allowed for studies of individual GnRH neurons. We have demonstrated previously the preservation of physiological function in male GnRH-GFP mice. In the present study, we confirm using biocytin-filled GFP-positive neurons in the hypothalamic slice preparation that GFP-expressing somata, axons, and dendrites in hypothalamic slices from GnRH-GFP rats are GnRH1 peptide positive. Second, we used repetitive sampling to study hormone secretion from GnRH-GFP transgenic rats in the homozygous, heterozygous, and wild-type state and between transgenic and Wistar males after ~4 yr of backcrossing. Parameters of hormone secretion were not different between the three genetic groups or between transgenic males and Wistar controls. Finally, we performed long-term recording in as many GFP-identified GnRH neurons as possible in hypothalamic slices to determine their patterns of discharge. In some cases, we obtained GnRH neuronal recordings from individual males in which blood samples had been collected the previous day. Activity in individual GnRH neurons was expressed as total quiescence, a continuous pattern of firing of either low or relatively high frequencies or an intermittent pattern of firing. In males with both intensive blood sampling (at 6-min intervals) and recordings from their GnRH neurons, we analyzed the activity of GnRH neurons with intermittent activity above 2 Hz using cluster analysis on both data sets. The average number of pulses was 3.9 ± 0.6/h. The average number of episodes of firing was 4.0 ± 0.6/h. Therefore, the GnRH pulse generator may be maintained in the sagittal hypothalamic slice preparation.     

Centrally administered orexin-A activates corticotropin-releasing factor-containing neurons in the hypothalamic paraventricular nucleus and central amygdaloid nucleus of rats: possible involvement of central orexins on stress-activated central CRF neurons

We examined the effects of centrally administered orexin-A on corticotropin-releasing factor (CRF)-containing neurons in the hypothalamic paraventricular nucleus (PVN) and the central amygdaloid nucleus (CeA) of rats, using dual immunostaining for CRF and Fos. Ninety minutes after intracerebroventricular administration of orexin-A, approximately 96% and 45% of CRF-containing neurons expressed Fos-like immunoreactivity (LI) in the PVN and the CeA, respectively. We also examined the effects of immobilized stress and cold exposure on orexin-A-containing neurons in the rat hypothalamus using dual immunostaining for orexin-A and Fos. After immobilized stress for 20 min and cold exposure at 4 degrees C for 30 min, approximately 24% and 15% of orexin-A-containing neurons expressed Fos-LI, respectively. These results suggest that orexins in the central nervous system may be involved in the activation of central CRF neurons induced by stress.     

Neurons of the Paraventricular Hypothalamic Nucleus Under Normal and Modified Illumination Conditions: Immunohistochemical and Morphometric Parallels

Using an immunohistochemical technique and densitometric analysis in experiments on rats, we examined the expression of an early-response protein, c-Fos, in the medial parvicellular subnucleus of the hypothalamic paraventricular nucleus (mpcPVN). During 7 days, the animals were kept under conditions of (i) normal photoperiod (group LD), (ii) continuous illumination (group LL), and (iii) continuous darkness (light deprivation, group DD). It was found that both normal photoperiodicity and artificial modifications of the latter result in significant changes of c-Fos immunopositivity in the nuclei of mpcPVN neurons (cells that synthesize corticotropin-releasing factor). Changes in the indices of concentration and amount of c-Fos in mpcPVN neurons were, from some aspects, similar to those observed under analogous experimental conditions in neurons of a leading pacemaker of the circadian periodicity, the suprachiasmatic nuclei (SChNs) of the hypothalamus [10], but, at the same time, demonstrated noticeable specificity from other aspects. In all experimental groups, the concentration and amount of c-Fos in mpcPVN neurons were greater during daytime than at night. Dramatic increases in the above parameters observed at daytime in rats subjected to light deprivation were the most significant effect of modifications of the photoperiod. Both continuous illumination and light deprivation resulted in some increase of the number (density) of c-Fos-immunopositive units in the mpcPVN. The nuclei of neurons of this structure demonstrated the phenomenon found earlier in SChN neuron; geometrical dimensions of the cell nuclei of these structures depended noticeably on both illumination conditions and periodicity. Probable mechanisms of the dependence of c-Fos expression in the hypothalamic nuclei on the photoperiod and its modifications, interdependence of such changes observed in the circadian pacemaker (SChN) and an important component of the system controlling the stress reactions (mpcPVN), and the relation of the above phenomena to the level of melatonin produced by the epiphysis cerebri (main neuroendocrine intermediator organizing the above-mentioned periodicity) are discussed.     

The proglucagon-derived peptide, glucagon-like peptide-2, is a neurotransmitter involved in the regulation of food intake

The dorsomedial hypothalamic nucleus harbors leptin sensitive neurons and is intrinsically connected to hypothalamic nuclei involved in feeding behavior. However, it also receives ascending input from the visceroceptive neurons of the brainstem. We have identified a unique glucagon-like-peptide-2 containing neuronal pathway connecting the nucleus of the solitary tract with the dorsomedial hypothalamic nucleus. A glucagon-like-peptide-2 fiber plexus targets neurons expressing its receptor within the dorsomedial hypothalamic nucleus. Pharmacological and behavioral studies confirmed that glucagon-like-peptide-2 signaling is a specific transmitter inhibiting rodent feeding behavior and with potential long-term effects on body weight homeostasis. The glucagon-like-peptide-1 receptor antagonist, Exendin (9-39) is also a functional antagonist of centrally applied glucagon-like-peptide-2.     

Correction of changes induced by toluene in the cortical and subcortical structures of albino rat brain

The number and weight of cells in the cortical and subcortical structures of the cerebral and cerebellar motor system in albino rats after a long-term exposure to toluene were determined. Toluene intoxication proved to kill projection neurons and interneurons in the sensorimotor cortex, ventrolateral thalamic nucleus, caudate nucleus, pallidum, red nucleus, and inferior olivary complex. The decreased number of cerebellar cells was mediated by atrophic changes as indicated by the decrease in the area and dry weight of Purkinje cells. The addition of plaferon LB to the diet attenuated the cytotoxic effect of toluene.     

Testosterone-induced alterations in the reactivity pattern of the neurons in the arcuate nucleus of male mice

The effects of testosterone administration on the hypothalamic arcuate nucleus were studied in adult male mice by means of karyometry. Four animals per group were sacrificed 1, 2, and 3 h after intramuscular injection of 100 micrograms testosterone propionate/100 g body weight. The nuclear diameter of neurons was measured in serial coronal sections. Testosterone induced an increase in the nuclear diameter of neurons located in the dorsal and medial periventricular zones of the arcuate nucleus. The neurons exhibiting the greatest changes in nuclear diameter were situated in the rostral portion of the nuclear area examined. In the central portion of the arcuate nucleus no response to testosterone was found. The present data support previous observations showing mosaically arranged nerve-cell groups in this hypothalamic nucleus.     

Constant light exposure aggravates POMC-mediated muscle wasting associated with hypothalamic alteration of circadian clock and SIRT1 in endotoxemia rats

Constant light exposure is widespread in the intensive care unit (ICU) and could increase the rate of brain dysfunction as delirium and sleep disorders in critical patients. And the activation of hypothalamic neuropeptides is proved to play a crucial role in regulating hypercatabolism, especially skeletal muscle wasting in critical patients, which could lead to serious complications and poor prognosis. Here we investigated the hypothesis that constant light exposure could aggravate skeletal muscle wasting in endotoxemia rats and whether it was associated with alterations of circadian clock and hypothalamic proopiomelanocortin(POMC) expression. Fifty-four adult male Sprague-Dawley rats were intraperitoneally injected with lipopolysaccharide(LPS) or saline, subjected to constant light or a 12:12?h light-dark cycle for 7 days. On day 8, rats were sacrificed across six time points in 24?h and hypothalamus tissues and skeletal muscle were obtained. Rates of muscle wasting were measured by 3-methylhistidine(3-MH) and tyrosine release as well as expression of two muscle atrophic genes, muscle ring finger 1(MuRF-1) and muscle atrophy F-box(MAFbx). The expression of circadian clock genes, silent information regulator 1(SIRT1), POMC and hypothalamic inflammatory cytokines were also detected. Results showed that LPS administration significantly increased hypothalamic POMC expression, inflammatory cytokine levels and muscle wasting rates. Meanwhile constant light exposure disrupted the circadian rhythm, declined the expression of SIRT1 as well as aggravated hypothalamic POMC overexpression and skeletal muscle wasting in rats with endotoxemia. Taken together, the results demonstrated that constant light exposure could aggravate POMC-mediated skeletal muscle wasting in endotoxemia rats, which is associated with alteration of circadian clocks and SIRT1 in the hypothalamus.     

哺乳、哺乳-禁乳及哺乳-禁乳后再哺乳对大鼠下丘脑胖素A免疫反应神经元的影响

为观察下丘脑胖素 A在哺乳期摄食增加和能量代谢中的作用,本研究采用脑连续切片之免疫组织化学和图像定量分析技术,对分娩后第 12 天非哺乳、持续哺乳、持续哺乳后禁哺乳过夜和持续哺乳 - 禁哺乳后再急性哺乳大鼠下丘脑胖素A免疫反应神经元的免疫反应性进行了观察和半定量分析。结果表明,分娩后持续哺乳 11 d, 大鼠的日摄食量较同期分娩的非哺乳大鼠明显增加(180%),一夜禁哺乳则明显降低哺乳大鼠的日摄食量(45%); 哺乳12 d, 大鼠下丘脑胖素 A免疫反应神经元的数目和平均染色强度较非哺乳大鼠明显增加(P<0.001,P<0.05); 禁哺乳过夜(15 h)明显降低哺乳大鼠胖素A免疫反应神经元的数目和平均染色强度(P<0.001,P<0.05),与非哺乳大鼠比较无明显差异;禁哺乳过夜后再急性哺乳2 h 明显增加禁哺乳大鼠胖素 A 免疫反应神经元的数目和平均染色强度(P<0.001,P<0.05),急性哺乳 5 h 后,虽亦明显增加禁哺乳大鼠胖素 A免疫反应性(P<0.05),但与急性哺乳 2 h 比较作用减弱。上述结果表明,持续哺乳和禁乳后再哺乳均导致下丘脑胖素A明显增加,提示哺乳期胖素A可能表达上调并可能与哺乳期摄食增加有关, 且吸乳动作与下丘脑胖素A样神经元之间可能存在某种神经或体液性联系途径。