The hypocretin neuron system: an arousal system in the human brain

Hypocretins are recently discovered neuropeptides produced by a small group of posterior hypothalamic neurons which project widely over the neuroaxis. In this study, we note that hypocretin neuron perikarya in the human brain are localized to the perifornical region of the posterior hypothalamus, extending into the lateral hypothalamus. These neurons lightly innervate all areas of cerebral cortex studied in a variable pattern with denser innervation of association cortex than primary motor or sensory cortex. There is a dense innervation of hypothalamus, locus coeruleus, raphe nuclei, midline thalamus and nucleus of the diagonal band-nucleus basalis complex of the forebrain. This pattern of projections from the hypocretin neurons is compatible with an important role in arousal and the maintenance of the waking state.     

Comparative Characterization of Glutamate Decarboxylase in Crude Homogenates of Oviduct, Ovary, and Hypothalamus

Some biochemical characteristics of L-glutamate decarboxylase (GAD) were compared using crude homogenates of the rat oviduct, ovary, and hypothalamus. As estimated by the measurement of CO2 production, the specific activities of oviductal and ovarian GAD were about 10 and 3% of the hypothalamic value, respectively. Stoichiometric formation of gamma-aminobutyric acid (GABA) and CO2 from L-glutamate could be observed in oviduct and hypothalamus, while in ovarian homogenates the production of CO2 was more than nine times that of GABA. Hypothalamic and tubal GAD showed similar time course, temperature dependence, and pH dependence. The dependence on protein concentration and on exogenous cofactor supply was also similar in these two tissues. The kinetic constant for L-glutamate as a substrate was nearly the same in oviduct (1.30 mM) and hypothalamus (1.64 mM). The responsiveness of tubal and hypothalamic GAD to various inhibitors was also similar. The present findings indicate that the oviductal and the hypothalamic GAD may be identical, and they suggest a possible GABAergic innervation of the Fallopian tube.     

Defective hypothalamic autophagy directs the central pathogenesis of obesity via the IkappaB kinase beta (IKKbeta)/NF-kappaB pathway

Autophagy has been recently demonstrated to control cell and tissue homeostasis, including the functions of various metabolic tissues. However, it remains unclear whether autophagy is critical for the central nervous system and particularly the hypothalamus for exerting metabolic regulation. Using autophagy-related protein 7 (Atg7) as an autophagic marker, this work showed that autophagy was highly active in the mediobasal hypothalamus of normal mice. In contrast, chronic development of dietary obesity was associated with autophagic decline in the mediobasal hypothalamus. To investigate the potential role of autophagy in the hypothalamic control of metabolic physiology, a mouse model was developed with autophagic inhibition in the mediobasal hypothalamus using site-specific delivery of lentiviral shRNA against Atg7. This model revealed that hypothalamic inhibition of autophagy increased energy intake and reduced energy expenditure. These metabolic changes were sufficient to increase body weight gain under normal chow feeding and exacerbate the progression of obesity and whole-body insulin resistance under high-fat diet feeding. To explore the underlying mechanism, this study found that defective hypothalamic autophagy led to hypothalamic inflammation, including the activation of proinflammatory IκB kinase β pathway. Using brain-specific IκB kinase β knockout mice, it was found that the effects of defective hypothalamic autophagy in promoting obesity were reversed by IκB kinase β inhibition in the brain. In conclusion, hypothalamic autophagy is crucial for the central control of feeding, energy, and body weight balance. Conversely, decline of hypothalamic autophagy under conditions of chronic caloric excess promotes hypothalamic inflammation and thus impairs hypothalamic control of energy balance, leading to accelerated development of obesity and comorbidities.     

Study of electrical activity in the area of the hypothalamic ventromedial nucleus and the lateral hypothalamus of rats fed on a high protein and high fat diet.

The authors studied bioelectric potentials in the area of the hypothalamic ventromedial nucleus and the lateral hypothalamus of rats fed on a standard, a high protein and a high fat diet. On the first 3--6 days after changing from the standard to the high fat and high protein diets, a decrease in the amplitude of electrical activity was recorded in both the areas in question. It was also found that the frequency of electrical activity in the hypothalamic ventromedial nucleus or the lateral hypothalamus rose, after 3 days administration of the high fat or the high protein diet, in correlation to the type of diet, and that, in the frequency spectrum, a change occurred in the proportion of basic frequency in relation to superimposed frequencies distorting it. It was further found that there was a permanent difference between the amplitude of electrical activity in the lateral hypothalamus and the hypothalamic ventromedial nucleus.     

The suprachiasmatic nucleus in the sheep: retinal projections and cytoarchitectural organization

The retinal innervation, cytoarchitectural, and immunohistochemical organization of the suprachiasmatic nucleus (SCN) was studied in the domestic sheep. The SCN is a large elongated nucleus extending rostrocaudally for roughly 3 mm in the hypothalamus. The morphology is unusual in that the rostral part of the nucleus extends out of the main mass of the hypothalamus onto the dorsal aspect of the optic chiasm. Following intraocular injection of wheat-germ agglutininhorseradish peroxidase or tritiated amino acids, anterograde label is distributed throughout the SCN. Retinal innervation of the SCN is bilaterally symmetric or predominantly ipsilateral. Quantitative image analysis demonstrates that, although the amount of autoradiographic label is greatest in the ventral and central parts of the nucleus, density varies progressively between different regions. In addition to the SCN, retinal fibers are also seen in the medial preoptic area, the anterior and lateral hypothalamic areas, the dorsomedial hypothalamus, the retrochiasmatic area, and the basal telencephalon. Whereas the SCN can be identified using several techniques, complete delineation of the nucleus requires combined tract tracing, cytoarchitectural, and histochemical criteria. Compared with the surrounding hypothalamic regions, the SCN contains smaller, more densely packed neurons, and is largely devoid of myelinated fibers. Cell soma sizes are smaller in the ventral SCN than in the dorsal or lateral parts, but an obvious regional transition is lacking. Using Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining, the SCN can be clearly distinguished in the rostral and medial regions, but is less differentiated toward the caudal pole. Immunohistochemical demonstration of several neuropeptides shows that the neurochemical organization of the sheep SCN is heterogeneous, but that it lacks a distinct compartmental organization. Populations of different neuropeptide-containing cells are found throughout the nucleus, although perikarya positive for vasoactive intestinal polypeptide and fibers labeled for methionine-enkephalin are predominant ventrally; neurophysine-immunoreactive cells are more prominent in the dorsal region and toward the caudal pole. The results suggest that the intrinsic organization of the sheep SCN is characterized by gradual regional transitions between different zones.     

Different distributions of nitric oxide synthase-containing neurons in the mouse and rat hypothalamus.

The distribution of nitric oxide synthase-containing neurons was studied in the rat and mouse hypothalamus by immunohistochemistry and NADPH-diaphorase histochemistry. Immunostaining and NADPH-diaphorase staining of hypothalamic neurons were comparable in all hypothalamic nuclei of either species except in the arcuate nucleus that stained positive for nitric oxide synthase immunoreactivity but negative for NADPH-diaphorase reactivity. The presence of nitric oxide synthase-immunopositive neurons in the arcuate nucleus was confirmed by nitric oxide synthase immunofluorescence viewed under the confocal microscope at 1 microm thickness. Cross-species comparison showed that, in general, the number and intensity of nitric oxide synthase-containing neurons were much higher in the rat than in the mouse hypothalamus. Differences in the distribution of nitric oxide synthase-containing neurons between these two rodents were found in most hypothalamic nuclei. In particular, two dense clusters of nitric oxide synthase-containing neurons were found in the paraventricular and supraoptic nuclei of the rat hypothalamus in contrast to their scarcity in the same nuclei of the mouse hypothalamus.     

Role of Catecholamines in Regulation of the Functional State of Vasopressinergic Cells of the Rat Hypothalamus

In in vivo and in vitro experiments there have been shown different mechanisms of regulation of hypothalamic vasopressinergic neurons, including regulation due to changes of activity level of brain catecholaminergic and NPY-ergic neurons innervating hypothalamic vasopressinergic cells. We demonstrated in in vitro experiments that dopamine and noradrenaline had no effects on vasopressin expression, but inhibited its release from cell perikarya in supraoptic and paraventricular nuclei of hypothalamus. Besides, activity of vasopressinergic neurons might probably be regulated via activation of synthesis of these neurotransmitters in vasopressinergic cells themselves in the supraoptic and paraventricular nuclei. To activate synthesis of various neurotransmitters, in our case, catecholamines and NPY, in vasopressinergic neurons, different stimuli adequate to trigger or activate synthesis of these substances are required. Synthesis of catecholamines in vasopressinergic cells of supraoptic and paraventricular nuclei was revealed after immobilization stress and adrenalectomy. NPY is synthesized in neurons of hypothalamic neurosecretory centers in norm, and its synthesis increases at disturbances of NPY-ergic innervation of vasopressinergic cells.     

Glucagon-related peptides in the rat hypothalamus

Summary Immunohistochemically, nerve fibers and terminals reacting with anti-N-terminal-specific but not with anti-C-terminal-specific glucagon antiserum were observed in the following rat hypothalamic regions: paraventricular nucleus, supraoptic nucleus, anterior hypothalamus, arcuate nucleus, ventromedial hypothalamic nucleus and median eminence. Few fibers and terminals were demonstrated in the lateral hypothalamic area and dorsomedial hypothalamic nucleus. Radioimmunoassay data indicated that the concentration of gut glucagon-like immunoreactivity was higher in the ventromedial nucleus than in the lateral hypothalamic area. In food-deprived conditions, this concentration increased in both these parts. This was also verified in immunostained preparations in which a marked enhancement of gut glucagon-like immunoreactivity-containing fibers and terminals was observed in many hypothalamic regions. Several immunoreactive cell bodies were found in the ventromedial and arcuate nuclei of starved rats. Both biochemical and morphological data suggest that glucagon-related peptides may act as neurotransmitters or neuromodulators in the hypothalamus and may be involved in the central regulatory mechanism related to feeding behavior and energy metabolism.     

Organization of hypothalamic area of diencephalon in the sturgeons

Cytoarchitectonics of hypothalamic area of diencephalon of the sturgeons was studied in serial sections by techniques of Nissl staining and Bielschowski impregnation in Viktorov’s modification. The hypothalamus was shown to be the most expanded area of diencephalon and forms its the most ventral part. The hypothalamic area of four studied sturgeons, the hausen, Huso huso L., Kura sturgeon, Acipenser guldenstaedtii persicus n. Kurensis Belyaeff, Caspian sturgeon, Ac. stellatus Pall. and the barbel sturgeon Ac. nudioventris Lov. was found to have similar structure. Eleven nerve structures are identified and described in the hypothalamic area: dorsal, ventral, and caudal periventricular zones, rostral and dorsal hypothalamic nuclei, ventral and ventrolateral hypothalamic nuclei, diffuse and central nuclei of the inferior lobes, nucleus of the vascular sac, and mammillary nucleus. Peculiarities and common features of organization of four major parts of hypothalamus of the sturgeons are considered in comparison with those of other ray-finned fish. The performed analysis indicates a high level of development of hypothalamus of the sturgeons.     

Unit responses in the anterior and posterior hypothalamus to stimulation of the splanchnic and sciatic nerves and to photic stimulation

By extracellular recording of spike discharges the sensory properties of neurons of the anterior and posterior regions of the cat hypothalamus were studied during stimulation of the splanchnic and sciatic nerves and during photic stimulation. Hypothalamic neurons were shown to be characterized by wide convergence of heterosensory excitation: 68% of spontaneously active hypothalamic neurons responded to somatovisceral and photic stimulation. Some posterior hypothalamic neurons responded to somatovisceral stimulation but not to photic stimulation. Neurons responding only to photic stimulation were found in the anterior hypothalamus; no neurons responding only to visceral stimulation were found in the hypothalamus. Total convergence of somatic and visceral afferentation of neurons of the posterior and anterior hypothalamus was observed. Mostly responses of phasic type were obtained to stimulation of all modalities. The study of the quantitative ratio between responses of excitatory and inhibitory types showed that the former predominate. The principles governing the functional organization of hypothalamic afferent systems are discussed.Academician L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 8, No. 3, pp. 276–282, May–June, 1976.     

Histamine-immunoreactive neurons in the hypothalamus of cats

The localization of histaminergic neurons in the cat brain was determined immunohistochemically with an antibody against histamine. We found that histamine-immunoreactive neurons are observed exclusively in the posterior hypothalamus of colchicine treated cats. The larger group of neurons was found in the ventrolateral part of the posterior hypothalamus, including the tuberomammillary nucleus. Histamine-positive neurons were also observed in the supramammillary area and adjacent posterior hypothalamic area, as well as in the peri- and premammillary regions. In addition, numerous histamine immunoreactive fibers were detected, not only in the posterior hypothalamus, but also in other brain areas, such as the preoptic area of the anterior hypothalamus.     

青紫蓝兔体内Ghrelin的免疫组化定位

采用免疫组织化学方法研究了Ghrelin在青紫蓝兔(Oryctolagus cuniculus)体内的分布定位。结果显示,Ghrelin免疫阳性细胞分布于下丘脑、大脑皮质、延髓、脊髓、胃、小肠和大肠。下丘脑内的阳性神经元胞体主要分布于弓状核、室旁核、腹内侧核、背内侧核和下丘脑外侧区。胃肠道内的Ghrelin免疫阳性细胞存在两种类型,即"闭合型"细胞和"开放型"细胞。实验结果表明,兔体内Ghrelin的分布与人和其他动物的分布基本相似,但也存在一些差异。     

Neural interaction of gonadotropin-regulating hormone immunoreactive neurons and the suprachiasmatic nucleus with the paraventricular organ in the Japanese grass lizard (Takydromus tachydromoides)

Our previous study demonstrated that the paraventricular organ (PVO) in the hypothalamus of the Japanese grass lizard (Takydromus tachydromoides) showed immunoreactivity against the light signal-transducing G-protein, transducin. This finding suggested that the PVO was a candidate for the deep-brain photoreceptor in this species. To understand functions of the PVO, we investigated distributions of transducin, serotonin, gonadotropin-releasing hormone (GnRH), and gonadotropin-inhibitory hormone (GnIH) in the lizard's brain. We immunohistochemically confirmed co-localization of transducin and serotonin in PVO neurons that showed structural characteristics of cerebrospinal fluid (CSF)-contacting neurons. GnRH-immunoreactive (ir) cells were localized in the posterior commissure and lateral hypothalamic area. Some of the serotonin-ir fibers extending from the PVO to the lateral hypothalamic area contacted the GnRH-ir cell bodies. GnIH-ir cells were localized in the nucleus accumbens, paraventricular nucleus, and upper medulla, and GnIH-ir fibers from the paraventricular nucleus contacted the lateral processes of serotonin-ir neurons in the PVO. In addition, we found that serotonin-ir fibers from the PVO extended to the suprachiasmatic nucleus (SCN), and the retrograde transport method confirmed the PVO projections to the SCN. These findings suggest that the PVO, by means of innervation mediated by serotonin, plays an important role in the regulation of pituitary function and the biological clock in the Japanese grass lizard.     

Brain natriuretic peptide (BNP)-like immunoreactivity in the central nervous system of the crested newt.

The distribution of brain natriuretic peptide (BNP)-like immunoreactivity (ir) was studied in the brain of a urodele amphibian, the crested newt Triturus carnifex Laur. BNP-like immunoreactive neurons were found mainly in the caudal hypothalamus (retro- and supra-chiasmatic areas) and in the preoptic area. A widespread innervation throughout the brainstem as far as the spinal cord was also observed. By double immunostaining (after section incubation with a-BNP and a-tyrosine hydroxylase-TH-antibodies), close topographical relationships between BNP-like and TH-like immunoreactive neurons within the hypothalamus were found.     

Distribution and quantification of immunoreactive orexin A in rat tissues.

A sensitive and specific radioimmunoassay for orexin A was developed. Orexin A immunoreactivity was found to be confined to the central nervous system (CNS) with the highest concentrations in the hypothalamus, inferior and superior colliculi and brainstem. Within the hypothalamus, the highest levels were found in the lateral and posterior hypothalamus. These regions had a greater orexin A content in females compared to males. The orexin A content of hypothalamic regions did not change with fasting and no difference was noted in hypothalami of rats fed a high fat diet. The hypothalamic orexin A content was not different in obese Zucker rats compared to lean controls. Thus, orexin A has a wide distribution in the CNS, but appetite regulation may not be its main function.     

Changes in the innervational connections of the urinary bladder upon experimental stimulation of the hypothalamus]

The innervation of the urinary bladder was studied in 40 rabbits, 5 of which were control and in 35 animals the posterior and anterior nuclei of the hypothalamus were stimulated by electric current through bipolar electrodes. The efferent innervation connections in whose terminal plexuses there occurred absorption of catecholamines were shown histochemically and neurohistologically to be involved in the pathological process. Reactive and degenerative changes were found to take place in receptory nerve terminations, and first of all in free ones. Then the bodies of pseudounipolar neurons of spinal ganglia were changed. Changes were also found in the connections of sensory neurons with the cells of Goll's and Burdach's nuclei.     

Nuclei and fields of the rabbit hypothalamus]

Common features and distinctions in the structure of certain fields of the rabbit hypothalamus were established on the basis of cytoarchitectonical and cytological analysis. It has been shown that there are three types of nerve cells in the nuclei and fields of the hypothalamus which can be referred to somato-, cyto- and karyochromic elements of the nervous system in accordance with the Nissl classification. All the cellular structures of the hypothalamus can be divided into heteromorphic and isomorphic types. The medial and lateral hypothalamic fields of all the rostro-caudal length of the hypothalamus are referred to the first type. The hypothalamic nuclei occupying its basal part are referred to the second type. On the basis of the obtained data concerning the neuronal composition of the fields and nuclei of the hypothalamus, it can be divided into three zones: the medial zone, including 3 medial hypothalamic fields disposed along the 3d ventricle; the lateral zone comprizing two a lateral hypothalamic fields occupying its lateral parts along its all length and the basal zone including hypothalamic nuclei disposed mainly in the ventral part of the hypothalamus.     

Responses of neurons of different hypothalamic structures to stimulation of tooth pulp and Aß afferents in the cat sciatic nerve

The response of neurons of different hypothalamic structures to stimulation of painful tooth pulp afferents and painless sciatic nerve Aß afferents was investigated during acute experiments on cats. It was found that 80.7%, 81.5%, and 71.4% of neurons of the posterior, tuberal, and anterior hypothalamus respectively, responded to stimulation of the tooth pulp. Shortest latency of response was recorded in the posterolateral hypothalamus. Latency of response was shorter in the lateral than in the medial structures throughout the hypothalamus. A distinct prevalence of excitatory response was found in neurons of the posterior area and an almost equal proportion of excitatory and inhibitory response in neurons of the tuberal and anterior hypothalamus. A high degree of convergence between noxious and nonnoxious somatic afferents were discovered in hypothalamic neurons: 85.8% of those studied responded to stimulation of the sciatic nerve Aß afferents. The comparable unidirectional response pattern of hypothalamic neurons to stimulation of tooth pump painful afferents and painless sciatic nerve Aß fibers point to the nonspecific nature of the response observed in the mainstream population of multisensory hypothalamic neurons. A small population of unimodal nociceptive neurons (14.2%) was found in the hypothalamus. Nociceptive responses of anterior hypothalamic neurons were distinguished by their long refractory phase, lasting 200–500 msec, and their low rate of reproduction during rhythmic stimulation of tooth pulp (1.5–2 Hz). Neuronal organization of the nociceptive hypothalamic afferent system is discussed together with the role of convergent and specific "nociceptive" neurons in the shaping of thalamic regulatory functions.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 18, No. 2, pp. 171–180, March–April, 1986.     

Stimulation of 17 beta-hydroxy steroid dehydrogenase in the rat anterior pituitary gland by progesterone

Anterior pituitary gland and hypothalamic 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) activity was measured in the immature castrated estradiol primed rat to determine if differences in enzyme activity could explain the progesterone induced reduction of bound estradiol nuclear receptors of the anterior pituitary gland but not the hypothalamus. Higher levels of 17 beta-HSD activity were found in the anterior pituitary gland as compared to the hypothalamus. The enzyme activity in the anterior pituitary gland was stimulated by progesterone administered either in combination with estradiol for 4 days or as a single injection following 4 days of estradiol priming. No progesterone effects were found on hypothalamic 17 beta-HSD. Under the experimental conditions used, progesterone administration did not alter uterine 17 beta-HSD. An increase in anterior pituitary gland and uterine 17 beta-HSD was also induced by estrogen administration.     

The regional distribution of adenylate cyclase in individual nuclei of rat hypothalamus

The regional distribution of adenylate cyclase (AC) activity in separated individual hypothalamic nuclei was studied. The highest activity was found in the paraventricular nucleus and the lowest activity in the median eminence. In several other hypothalamic nuclei, there was also found higher activity of AC than in the total hypothalamus. These data show that in the individual hypothalamic nuclei there exist considerable differences in the activity of adenylate cyclase that may indicate their different function in regulation of neuroendocrine phenomena.