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An autoradiographic study of the efferent connections of the lateral hypothalamic area in the rat.

The efferent connections of the lateral hypothalamic area (LHA) have been analyzed in a series of 30 rat brains with injections of 3H-amino acids into different parts of the area and the surrounding regions. Our findings indicate that all parts of the LHA contribute ascending and descending fibers to the medial forebrain bundle, and also project medially to certain of the adjoining hypothalamic nuclei. All levels of the LHA appear to send some fibers to a continuous group of structures that extends from the medial septal-diagonal band complex rostrally, through the lateral preoptic and lateral hypothalamic areas to the mammillary complex and the ventral tegmental area caudally. In addition, it is evident that cells at different levels within the LHA may have differential projections. Thus, the anterior and lateral parts of the LHA also appear to project substantially to the anterior hypothalamic area, the ventromedial and dorsomedial hypothalamic nuclei, the parataenial and paraventricular nuclei of the thalamus, and the medial part of the lateral habenular nucleus. Similarly, cells in the tuberal and posterior parts of the LHA project to the central gray, the longest projections from the posterior region reaching as far caudally as the central tegmental field, the parabrachial nucleus, the locus coeruleus, and the superior central and dorsal nuclei of the raphe. Viewed as a whole, the LHA is therefore well-suited to integrate inputs from the limbic system and brainstem and to relay them on the one hand to the medial zone of the hypothalamus and on the other to virtually every structure closely associated with the medial forebrain bundle and to the nuclei of origin of the major ascending monoaminergic systems.

Animals↗

Increased neuropeptide Y concentrations in the lateral hypothalamic area of the rat after the onset of darkness: possible relevance to the circadian periodicity of feeding behavior.

Neuropeptide Y (NPY) is a major hypothalamic peptide which powerfully stimulates feeding when injected into the hypothalamus and is implicated in circadian rhythmicity. To investigate whether NPY is involved in the increased feeding that follows the onset of darkness in rats, NPY levels were measured in discrete hypothalamic areas before and after darkness. Four groups of eight adult female Wistar rats were habituated to a 12:12 hour light:dark cycle, with food presented at the onset of darkness (19.00 hours). One group was sacrificed during the 3 hours before darkness and another in the first 2.5 hours after darkness, with food provided as usual. To distinguish any effects of feeding itself, the study was repeated with two further groups, but food was not provided after darkness. Seven hypothalamic regions were microdissected from slices of fresh brain and acid-extracted for radioimmunoassay of NPY. NPY levels (fmol/microgram protein) were significantly higher (p less than 0.01) in the lateral hypothalamic area (LHA) of the dark-phase group in both studies. In the other six regions, NPY levels did not differ between light and dark phases. The LHA regulates the circadian rhythmicity of feeding and NPY injection here stimulates feeding. Alterations in NPY in the LHA around the onset of darkness may be related to the initiation of dark-phase feeding.

Animals↗

Estrogen decreases the responsiveness of subfornical organ neurons to angiotensinergic neural inputs from the lateral hypothalamic area in the female rat.

Twenty-eight subfornical organ (SFO) neurons in ovariectomized (OVX) female rats that were treated with propylene glycol (PG) vehicle and 26 SFO neurons in OVX female rats that were treated with estrogen benzoate (EB) were antidromically activated by electrical stimulation of the hypothalamic paraventricular nucleus (PVN) under urethane anesthesia. No significant differences were observed between the PG-treated and EB-treated OVX animals in the latency, conduction velocity, or threshold of antidromic activation. The mean spontaneous discharge rate was significantly lower in the EB-treated than in the PG-treated OVX animals. In both groups, the activity of the majority (86% in the PG-treated animals and 88% in the EB-treated animals) of identified SFO neurons were activated by microiontophoretic application of angiotensin II (ANG II). Electrical stimulation of the lateral hypothalamic area (LHA) increased the excitability of these ANG II-sensitive SFO neurons (58% in the PG-treated animals and 52% in the EB-treated animals). The excitatory response to either ANG II or LHA stimulation was blocked by microiontophoretic application of the ANG II antagonist saralasin (Sar), suggesting that the excitatory response to LHA stimulation may be mediated by angiotensinergic LHA projections to the SFO. The magnitude of excitatory response to either ANG II or the LHA stimulation was much greater in the PG-treated than in the EB-treated animals. These results suggest that estrogen decreases the responsiveness of SFO neurons projecting to the PVN to angiotensinergic inputs from the LHA.

Angiotensin II↗

Water deprivation increases the expression of neuronal nitric oxide synthase (nNOS) but not orexin-A in the lateral hypothalamic area of the rat.

The discovery that the lateral hypothalamic area (LHA) might be important in modulating drinking behavior and fluid balance has led to numerous studies aimed at identifying the key neurotransmitters/neuromodulators and pathways involved. While past studies have demonstrated the presence of neuronal nitric oxide synthase (nNOS) within the LHA, its role in the regulation of fluid homeostasis is not known. In light of this, and the mounting evidence suggesting a role for nitric oxide in osmotic regulation within the hypothalamus, this study sought to determine the effects of 24- and 72-hours of water deprivation on nNOS protein expression within the LHA of the rat with immunohistochemistry. In euhydrated control animals we observed nNOS-like immunoreactivity throughout all levels of the LHA. Following 24 hours of dehydration the number of nNOS-like immunopositive neurons was significantly increased in the rostral but not the caudal regions of LHA. Seventy-two hours of water deprivation lead to further increases in nNOS-like immunoreactivity at different levels of the LHA. Interestingly, however, we observed increased nNOS-like immunoreactivity in the caudal regions of the LHA that was not evident after 24 hours of water deprivation. Double-labeling immunofluorescence histochemistry revealed that the nNOS-like immunoreactive neurons were not colocalized with the orexin-A-containing neurons. These results suggest that an osmotic challenge leads to an upregulation of nNOS immunoreactivity within discrete areas of the LHA. This altered neurochemistry within the LHA further highlights the potential importance of nitric oxide and the LHA in central regulation of fluid homeostasis.

Animals↗

Morphometry of cytoarchitecture of the lateral hypothalamic area in the rat.

The cytoarchitecture of the lateral hypothalamic area (LHA) in rats was studied with cresyl violet-stained coronal celloidin sections and with sections of the brain impregnated by a Golgi method. Unimodality was established in the frequency distribution histogram of both the somatic cross-sectional area of the LHA neurons and somatic shape (elongation and circularity). The predominant somatic orientation was in the dorsomedial-ventrolateral direction: bimodality of the frequency distribution of somatic orientation was denied. These findings suggest that the LHA neurons examined in the present study are not subdivisible on the basis of the somatic area, shape or orientation. Although the neurons were classified into eight types based upon the dendritic pattern, those in the LHA largely consisted of only three of them; Type III (dendrites extending in two directions along the long axis of soma), Type IV (three directions) and Type VIII (four directions) jointly accounted for 97.1 percent of the total neurons examined. This finding suggests that the parameter of dendritic pattern serves an important purpose in the typing of the rat LHA neurons. The orientation of intrinsic dendrites and intrinsic axons in the LHA has also been described.

Animals↗

[Ultrastructure of neurons and synapses of the lateral hypothalamic area in the rat].

Ultrastructural organization of the lateral hypothalamic area (LHA) has been studied in Wistar rats. Structural peculiarities of neurons and interneuronal connections in the anterior, middle and posterior parts of the LHA have been demonstrated. In the anterior and middle parts, neurons of middle size (17-22 mcm) make the most of the cellular population; large neurons (35-50 mcm) predominate in the posterior part of the LHA. Small cells are diffusely distributed in all the parts. Two types of axons are defined: axons of the I type--with round light, and some of them, with large granular synaptic vesicles; axons of the II type contain smaller and polymorphous light synaptic vesicles. The axons form axodendritic and axosomatic synapses. The former predominate in all parts of the LHA. The axons of the I type form, mainly, asymmetrical, and the axons of the II type--symmetrical contacts. Most of the synapses are formed by the axons of the I type, overwhelmed majority of the synapses are axodendritic. Differences in localization and in distribution of the axodendritic synapses in different parts of the LHA are revealed. In the anterior part of the LHA the axodendritic synapses form synaptic muffs around large and middle dendrites, and more seldom around small and spine-shaped processes of the dendrites. In the middle and posterior parts of the LHA no synaptic muffs are revealed.

Animals↗

A topographical analysis of the origin of some efferent projections from the lateral hypothalamic area in the rat.

Some projections from the lateral hypothalamic area in the rat have been investigated, using combinations of fluorescent tracers, injected into several different parts of the central nervous system. Projections appear to arise from loosely organized assemblies of neurons, called sets and from more densely packed assemblies, called clusters. The sets and clusters vary considerably in position and in distinctness of their borderlines. Even within extensive and vaguely defined sets, however, high concentrations of labeled neurons may be present at specified sites in the lateral hypothalamus. Such concentrations are observed in the transitional area of the zona incerta and the dorsal part of the lateral hypothalamus, and in the ventrolateral part of the hypothalamus, bordering the cerebral peduncle and the subthalamic nucleus, in both cases after injections into some "autonomic centers" in the brainstem, such as the parabrachial nuclei and the dorsal vagal complex. Sets and clusters may overlap considerably. Within the fields of overlap the number of double labeled neurons may vary from almost zero up to more than 50%, depending on the injection sites. The results show that different parts of the lateral hypothalamus in the rat have different efferent relationships. Combination of the results of the present study with known data concerning the afferent relationships, the cytoarchitecture and behavioral functions of the lateral hypothalamic area, suggests that different parts of this entity are involved in different regulatory and behavioral functions.

Animals↗

Angiotensin II-sensitive neurons in the rat lateral hypothalamic area with efferent projections to the subfornical organ.

Thirteen neurons in the lateral hypothalamic area were antidromically activated by electrical stimulation of the subfornical organ in the rat. The activity of almost all identified neurons (N = 11) was excited by microiontophoretically applied angiotensin II. On the other hand, of the antidromically unidentified lateral hypothalamic area neurons (N = 10) tested, 4 were excited by microiontophoretically applied angiotension II and 6 were not affected. The excitatory responses of identified and unidentified lateral hypothalamic area neurons to angiotensin II were blocked by microiontophoretically applied saralasin, an angiotensin II antagonist.

Action Potentials↗

Leptin differentially regulates NPY and POMC neurons projecting to the lateral hypothalamic area.

Recent studies have reinforced the view that the lateral hypothalamic area (LHA) regulates food intake and body weight. We identified leptin-sensitive neurons in the arcuate nucleus of the hypothalamus (Arc) that innervate the LHA using retrograde tracing with leptin administration. We found that retrogradely labeled cells in the Arc contained neuropeptide Y (NPY) mRNA or proopiomelanocortin (POMC) mRNA. Following leptin administration, NPY cells in the Arc did not express Fos but expressed suppressor of cytokine signaling-3 (SOCS-3) mRNA. In contrast, leptin induced both Fos and SOCS-3 expression in POMC neurons, many of which also innervated the LHA. These findings suggest that leptin directly and differentially engages NPY and POMC neurons that project to the LHA, linking circulating leptin and neurons that regulate feeding behavior and body weight homeostasis.

Animals↗

Metabolic and neuroendocrine indices one month after lateral hypothalamic area lesions.

Mature male rats received bilateral electrolytic lateral hypothalamic area lesions (LHAL) or were sham-operated fed ad lib (CON-ADLIB) or sham-operated pair-fed/gained (CON-PF) to LHAL rats. One month later all rats were sacrificed. Rats with LHAL were hypophagic and had reduced body carcass fat, testes, livers, epididymal fat pads (PADS), diaphragms (DIA), and body weight compared to CON-ADLIB. In the liver, LHAL rats incorporated more C14-U-glucose carbon (GLUCINC) into lipid and glycogen than both CON groups, but GLUCINC was similar among CON groups. In PADS, LHAL rats oxidized more glucose carbon (GLUCOX) than CON-ADLIB but less than CON-PF/PG. The latter showed greater GLUCOX than CON-ADLIB. In DIA, LHAL and CON-PF/PG showed reduced GLUCINC into glycogen vs. CON-ADLIB. Plasma glucose was similar among groups, but insulin was lower in LHAL and CON-ADLIB than in CON-PF/PG. Rats with LHAL had lower plasma T3 concentrations than CON-ADLIB, but similar T3 levels compared to CON-PF/PG. Several of the metabolic changes in LHAL rats could be due to hypophagia; however, four out of nine metabolic indices, glucose carbon incorporation into liver lipid and glycogen and epididymal fat pad lipid and oxidation, were significantly different from CON-PF/PG, i.e., they were independent of food intake. Possibly then, they are due to a lesion effect other than on feeding mechanisms. Some aspects of metabolism that were previously found to be altered 48 h after LHAL were recovered, whereas others apparently were not.

Adipose Tissue↗

The region of the pontine parabrachial nucleus is a major target of dehydration-sensitive CRH neurons in the rat lateral hypothalamic area.

Neurons in a restricted part of the lateral hypothalamic area (LHA) show increased expression of corticotropin-releasing hormone (CRH) mRNA as a consequence of cellular dehydration. In the present study, we have investigated the organization of their efferent projections by using anterograde and retrograde tracing techniques. Additionally, we have compared the distribution of CRH mRNA-containing neurons after cellular dehydration and intraventricular (i.c.v.) colchicine injections. Our results show that cellular dehydration activates a more restricted neuronal population than does i.c.v. colchicine. Iontophoretic injections of Phaseolus vulgaris leucoagglutinin (PHAL) were placed in the LHA of animals drinking hypertonic saline and their proximity to activated CRH neurons determined by in situ hybridization for CRH mRNA. Although labelled fibers from these injections were seen throughout the brain, the region of the parabrachial nucleus and nucleus of the solitary tract (NTS) were most conspicuous in also having CRH immunoreactive fibers. Injections of Fluoro-Gold placed in these two structures were used to confirm these findings in dehydrated animals. Significant numbers of neurons containing both Fluoro-Gold and CRH mRNA were seen in the lateral hypothalamus after injections in the lateral and medial parts of the parabrachial nucleus; far fewer were seen after injections in the NTS. These results strongly suggest that the CRH neurons in the LHA activated by cellular dehydration provide an input to the region of the parabrachial nucleus. The altered biochemical composition of this pathway may well be able to modify sensory and motor patterns both during and after dehydration.

Animals↗

Inhibition of renal electrolyte excretion by gabaergic pathways of the lateral hypothalamic area.

Injection of gamma amino butyric acid (GABA) into the lateral hypothalamic area of unrestrained conscious rats caused a decrease in renal electrolyte excretion with an increase in urinary flow. When picrotoxin, a specific inhibitor of gabaergic pathways, was administered, a significant increase in renal water and electrolyte excretion occurred. The effect of simultaneous injection of picrotoxin and GABA into the same site indicated that picrotoxin was less potent in reversing the effect induced by GABA than GABA was in reversing the effect of picrotoxin. We conclude that GABA acts directly on the neuronal mechanisms involved in the control of water and electrolyte excretion, perhaps by exerting a tonic inhibitory action on renal electrolyte excretion.

Animals↗

Cardiovascular effects produced by L-glutamate stimulation of the lateral hypothalamic area.

L-Glutamate microinjections into the tuberal region of the lateral hypothalamic area (LHAt) caused a fall in blood pressure and heart rate in pentobarbital-anesthetized rats. The bradycardia was mediated by both beta-adrenergic and muscarinic mechanisms as demonstrated with pharmacological blockade. The hypotension was due to a decrease in cardiac output, not a decrease in total peripheral resistance. In addition, there was a reduction in coronary blood flow. If heart rate was held constant by pharmacological blockade or by electrical cardiac pacing, L-glutamate stimulation of the LHAt still caused a fall in blood pressure. When the electrically paced model was used, this hypotension was due to a fall in cardiac output. In contrast, with the pharmacological blockade of the heart, the hypotension was due to a decrease in the total peripheral resistance. The cardiac output reduction in the paced condition was not mediated solely by either beta-sympathetic or parasympathetic mechanisms as determined by pharmacological blockade. With heart rate held constant by either drugs or pacing, LHAt stimulation did not alter regional blood flow or resistance in any vascular bed, including the coronary circulation. We conclude that L-glutamate stimulation of the LHAt lowers the cardiac output and heart rate by both parasympathetic and beta-adrenergic mechanisms and elicits hypotension by lowering cardiac output in the naive and electrically paced model.

Animals↗

In vitro [U-14C]glucose utilization by tissues of weanling rats with lateral hypothalamic area lesions one month after lesion production.

The role of the lateral hypothalamic area (LHA) in intermediary metabolism was investigated by quantitation of [U-14C]glucose oxidation to 14CO2 and 14C incorporation into the glycogen and lipid fraction of the liver, epididymal fat pad, and diaphragm. Weanling male Sprague-Dawley rats received bilateral electrolytic lesions in the LHA (LHAL rats). Sham operated rats were either fed ad libitum (CON-ADLIB) or pair-gained to the LHAL rats (CON-PG). The experiment was terminated 1 month after lesion production. LHAL rats were significantly (SIG) lighter and shorter and ate less than CON-ADLIB; LHAL rats were also SIG shorter than CON-PG, pointing to a food intake-independent lesion effect. Both LHAL and CON-PG rats had SIG less percent carcass fat than CON-ADLIB, but there was no SIG difference between LHAL and CON-PG rats. Also, LHAL rats had a SIG higher percentage of carcass protein than both CON-ADLIB and CON-PG. Furthermore, LHAL rats incorporated SIG less glucose into liver glycogen than CON-ADLIB but SIG more into CON-PG, whereas CON-PG rats incorporated SIG less into liver glycogen than CON-ADLIB, again suggesting a food intake-independent effect. There was no difference among the groups in glucose oxidation and incorporation into lipids and glycogen in both diaphragm and epididymal fat pads and liver total lipid. However, livers of CON-PG metabolized SIG more [U-14C]glucose to CO2 than did livers of CON-ADLIB, suggesting a food intake-dependent effect. There was no difference between LHAL and CON-PG rats in this parameter.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

Changes in urinary electrolytes excretion after injections of ionic solutions into lateral hypothalamic area of rats.

Rats submitted to a chronically implanted cannula into lateral hypothalamic area (LHA) were utilized to study the effect of ionic stimulation of KCl, LiCl and CaCl2, sodium free solutions, on renal water and electrolyte excretion. In a general way decreases in natriuresis and kaliuresis following the injections of solutions into LHA were observed. Furthermore, significant increases in diuresis were noted after the injections of the 0.30 M solutions.

Animals↗

Distinct patterns of neuropeptide gene expression in the lateral hypothalamic area and arcuate nucleus are associated with dehydration-induced anorexia.

We have investigated the hormonal and hypothalamic neuropeptidergic substrates of dehydration-associated anorexia. In situ hybridization and hormone analyses of anorexic and paired food-restricted rats revealed two distinct profiles. First, both groups had the characteristic gene expression and endocrine signatures usually associated with starvation: increased neuropeptide Y and decreased proopiomelanocortin and neurotensin mRNAs in the arcuate nucleus (ARH); increased circulating glucocorticoid but reduced leptin and insulin. Dehydrated animals are strongly anorexic despite these attributes, showing that the output of leptin- and insulin-sensitive ARH neurons that ordinarily stimulate eating must be inhibited. The second pattern occurred only in anorexic animals and had two components: (1) reduced corticotropin-releasing hormone (CRH) mRNA in the neuroendocrine paraventricular nucleus (PVH) and (2) increased CRH and neurotensin mRNAs in the lateral hypothalamic (LHA) and retrochiasmatic areas. However, neither corticosterone nor suppressed PVH CRH gene expression is required for anorexia after dehydration because PVH CRH mRNA in dehydrated adrenalectomized animals is unchanged from euhydrated adrenalectomized controls. We also showed that LHA CRH mRNA was strongly correlated with the intensity of anorexia, increased LHA CRH gene expression preceded the onset of anorexia, and dehydrated adrenalectomized animals (which also develop anorexia) had elevated LHA CRH gene expression with a distribution pattern similar to intact animals. Finally, we identified specific efferents from the CRH-containing region of the LHA to the PVH, thereby providing a neuroanatomical framework for the integration by the PVH of neuropeptidergic signals from the ARH and the LHA. Together, these observations suggest that CRH and neurotensin neurons in the LHA constitute a novel anatomical substrate for their well known anorexic effects.

Adrenalectomy↗

Acetylcholinesterase-containing cells in the lateral hypothalamic area are immunoreactive for alpha-melanocyte stimulating hormone (alpha-MSH) and have cortical projections in the rat.

Small injections of the retrograde fluorescent tracer Fast blue into different cortical areas, including the hippocampal region, labeled cells in the zona incerta and the lateral hypothalamic area. A majority (approximately equal to 90%) of the retrogradely labeled cells cross-reacted with an anti-serum to the opioid peptide alpha-melanocyte stimulating hormone (alpha-MSH). Sequential staining of the same tissue sections showed that a majority of the alpha-MSH stained cells in the lateral hypothalamic area and the zona incerta also contain the enzyme acetylcholinesterase (AChE) but not cholineacetyltransferase. These results suggest that cortical AChE resides partly in non-cholinergic terminals and that some of these arise from alpha-MSH immunoreactive cells in the hypothalamus and subthalamus.

Acetylcholinesterase↗

[The functional connection of rat habenular nucleus and lateral hypothalamic area in the regulation of cardiovascular activities].

The electrical stimulation of the lateral hypothalamic area (LH) in the rat elicited an obvious rise of the arterial pressure and increase in the heart rate. After the microinjection of lidocaine hydrochloride into the habenular nucleus (Hb) on the same side of the electrode, the vasopressor effect elicited by the electrical stimulation of the LH was blocked by 38.9% and the effect on heart rate was reduced by 44.4%. After the microinjection of lidocaine hydrochloride into the bilateral Hb, the vasopressor effect elicited by the electrical stimulation of the LH was blocked by 40.7% and the effect on heart rate was reduced by 41.2%. After the microinjection of the artificial cerebrospinal fluid into the unilateral or bilateral Hb, the cardiovascular effect elicited by the electrical stimulation of the LH was not affected. The electrical stimulation of the Hb in the rat elicited an obvious rise of the arterial pressure, but the heart rate was not changed significantly. After the microinjection of lidocaine hydrochloride into the LH on the same side of the electrode the vasopressor effect elicited by the electrical stimulation of the Hb was blocked by 63.2%. After the microinjection of lidocaine hydrochloride into the bilateral LH the vasopressor effect elicited by the electrical stimulation of Hb was blocked by 62.6%. After the microinjection of the artificial cerebrospinal fluid into the unilateral or bilateral LH, the cardiovascular effect elicited by the electrical stimulation of the Hb was not affected. The present study shows that the Hb and LH exert synergistic action in the regulation of the cardiovascular activities.

Animals↗