Lateral and ventromedial hypothalamic influences on hepatic autonomic nerve activity in the rat

Effects of hypothalamic stimulation and lesion on hepatic autonomic nerve activity were investigated in anesthetized rats. Stimulation of the lateral hypothalamic area increased activity of the hepatic vagal nerve and decreased activity of the splanchnic nerve. Lesion of this area produced a rapid and strong reduction of vagal nerve activity and induced either an increase or a decrease of splanchnic nerve activity. In contrast, stimulation of the ventromedial hypothalamic nucleus produced a strong inhibition of vagal nerve activity and facilitation of splanchnic nerve activity. However, lesion of this nucleus facilitated or inhibited activities in both nerve branches. The functional significance of the relationship between the hypothalamus and the hepatic autonomic nerve in regulation of glucose metabolism in the liver are discussed.     

Effects of hypothalamic lesion on pancreatic autonomic nerve activity in the rat

The effects of hypothalamic lesions and intravenous glucose infusion on the efferent activity of vagal and splanchnic nerves to the pancreas were studied in anesthetized rats. Lesions of the ventromedial hypothalamic (VMH), the dorsomedial hypothalamic (DMH) and the paraventricular (PVN) nuclei increased vagal and reduced splanchnic nerve activity. Lesion of the lateral hypothalamic area (LHA) decreased pancreatic vagal nerve activity, and produced either increased or decreased activity of pancreatic splanchnic nerve. Intravenous glucose infusion increased activity of the vagal nerve and reduced that of the splanchnic nerve. These glucose responses were influenced by hypothalamic lesions only slightly or not at all. The findings suggest that hypothalamic modulation of pancreatic hormone secretion involves both the parasympathetic and sympathetic nervous systems, and provide evidence that not only the VMH and the LHA but also the DMH and the PVN are involved in this mechanism.     

NOS isoenzyme content in brain nuclei as related to food intake in experimental cancer cachexia

Evidence implies that nitric oxide (NO) in the central nervous systems mediates anorexia in tumor-bearing hosts. We have therefore evaluated, by immunohistochemical image analyses, net alterations of nitric oxide synthases (nNOS, eNOS, iNOS) in brain nuclei [paraventricular hypothalamic nucleus (PVN), medial habenular nucleus (MHB), lateral habenular nucleus (LHB), paraventricular thalamic nucleus (PV), lateral hypothalamic area (LHA), ventromedial hypothalamic nucleus (VMH), nucleus of the solitary tract (NTS)] of tumor-bearing mice (TB) with prostanoid-related anorexia. Pair-fed (PF) and freely fed (FF) non-tumor-bearing mice were used as controls. c-fos was analyzed as indicator of neuronal activation. nNOS was significantly increased in VMH and PVN from TB mice, while eNOS was significantly increased in LHB and LHA. iNOS was significantly increased in LHA and PVN nuclei, but decreased in MHB, LHB and VMH from tumor-bearers. However, several of these alterations were similarly observed in brain nuclei from pair-fed controls. Provision of unspecific NOS-antagonists to TB mice increased nNOS, eNOS and iNOS in several brain nuclei (PVN, LHA, VMH), but left tumor-induced anorexia unchanged. c-fos was significantly increased in all brain nuclei in PF mice except for NTS, LHA and PVN compared to controls, while tumor-bearing mice had increased c-fos in LHA and PVN only compared to controls. Our results demonstrate a complex picture of NOS expression in brain areas of relevance for appetite in tumor-bearing hosts, where most changes seemed to be secondary to stress during negative energy balance. By contrast, NOS content in PVN and LHA nuclei remains candidate behind anorexia in tumor disease. However, nitric oxide does not seem to be a primary mediator behind tumor-induced anorexia. NO may rather secondarily support energy intake in conditions with negative energy balance.     

Vagal and splanchnic effects at the level of the ventromedian nucleus of the hypothalamus (VMH) in the cat

A systematic study of vagal and splanchnic projections to the VMH area was undertaken in anaesthetized cats by means of macroelectrodes and microelectrodes. Responses elicited by vagal and splanchnic nerve stimulation were recorded from the VMH nucleus and the region situated above it. The fact that they are identical, whatever the nerve stimulated, indicates that the afferences follow a common central pathway. From the response latency and the stimulation parameters, it is assumed that the small vagal and splanchnic fibres (Aδ, B and C) are involved. Evoked potentials are generally constituted of two parts: the earlier implies certainly the VPL nucleus (since it disappears definitively after VPL coagulation), whereas the later concerns a different relay probably located in the neighbouring and associative structure (since it persists after VPL coagulation). Similar late responses were simultaneously recorded in VPL nucleus, suggesting that this structure was the same for both nuclei. Its exact location was discussed.On the other hand, effects of gastric distension on the evoked potentials produced by vagal and splanchnic nerve stimulation were studied. The results obtained (decrease or increase in the responses, occurrence of an additional potential) indicated clearly that the gastric afferences projected to the VMH region. This simple method could be used to determine the importance of the sensory innervation of the different parts of the digestive tract.     

Acetylcholinesterase activity changes on visceral organs of VMH lesion-induced obese rats

It is accepted that the tone of the parasympathetic nervous system increases after VMH lesion, whereas the sympathetic tone decreases. To reinforce investigations over outcomes from disturbances of the hypothalamic neuronal systems on peripheral autonomic nerve activity this study determined the acetylcholinesterase (AchE) activity in visceral organs, known as vagal targets, from VMH-lesioned obese rats. It was found that AchE activity was significantly increased in liver, pancreas, and stomach from these animals. However, it was not changed in kidneys, being decreased in spleen. The results suggest that AchE activity is enhanced in vagus innervated tissues to following up the unbalance of the autonomic nervous system as observed in VMH lesion-induced obesity.     

ACETYLCHOLINESTERASE ACTIVITY CHANGES ON VISCERAL ORGANS OF VMH LESION–INDUCED OBESE RATS

It is accepted that the tone of the parasympathetic nervous system increases after VMH lesion, whereas the sympathetic tone decreases. To reinforce investigations over outcomes from disturbances of the hypothalamic neuronal systems on peripheral autonomic nerve activity this study determined the acetylcholinesterase (AchE) activity in visceral organs, known as vagal targets, from VMH-lesioned obese rats. It was found that AchE activity was significantly increased in liver, pancreas, and stomach from these animals. However, it was not changed in kidneys, being decreased in spleen. The results suggest that AchE activity is enhanced in vagus innervated tissues to following up the unbalance of the autonomic nervous system as observed in VMH lesion–induced obesity.     

Distribution of prodynorphin mRNA and its interaction with the NPY system in the mouse brain

Using radioactive in situ hybridisation, the distribution of prodynorphin mRNA in the brains of C57Bl/6 mice was systemically investigated, and double-labelling in situ hybridisation was used to determine the extent to which neuropeptide Y (NPY) and prodynorphin mRNAs were co-expressed. Our results demonstrate that prodynorphin mRNA expression in the mouse brain is localised at specific subregions of the olfactory bulb, cortex, hippocampus, amygdala, basal ganglia, thalamus, hypothalamus, mesencephalon and myelencephalon. Among the regions displaying the most intense labelling were the olfactory tubercle, lateral septum (LS), caudate putamen (Cpu), central amygdaloid nucleus (Ce), paraventricular hypothalamic nucleus (PVN), supraoptic nucleus (SO), lateral hypothalamic area (LHA), ventromedial hypothalamic nucleus (VMH), lateral reticular nucleus (LRt) and solitary tract nucleus (NTS). In the arcuate nucleus of the hypothalamus (Arc), double-labelling in situ hybridisation revealed that prodynorphin expressing neurons also contained NPY mRNA, with a co-localisation rate of approximately 88% in the lateral part of the Arc, and 79% in the dorsal part of the Arc, respectively, suggesting potential overlapping functions of these two neurotransmitters in feeding type behaviour.     

Hypothalamic control of liver glycogen metabolism in adult and aged rats

The regulatory function of the hypothalamus in the metabolism of liver glycogen was investigated by analyzing the changes in the activities of the rate-limiting enzymes implicated in glycogen breakdown (glycogen phosphorylase) and synthesis (glycogen synthetase), after electrical stimulation of the ventromedial hypothalamic nucleus (VMH) and the lateral hypothalamic nucleus (LH) of rats. Electrical stimulation of the VMH induced rapid and marked increase in the content of the active form of phosphorylase in the liver, but it did not affect the synthetase. Stimulation of the LH, on the other hand, caused an increase in the level of the active form of synthetase, but produced little change in the phosphorylase activity. The concentration of insulin in the portal blood decreased significantly on stimulation of the VMH, but did not change appreciably on stimulation of the LH. The ability of the hypothalamus to control liver glycogen metabolism was found to be impaired in 2-year-old rats: the magnitude and duration of the early increase in active phosphorylase in response to VMH stimulation were reduced, and the response of synthetase to LH stimulation was slow.The results are discussed from the viewpoint that the VMH and LH are the centers for controlling irect neural and neural-hormonal regulations of liver glycogen metabolism. It is concluded that the VMH-splanchnic nerve pathway is an important neural component for controlling glycogenolysis and glucose output by the liver, and that the LH-vagal pathway is an opposing neural component for controlling glycogenesis in the liver.     

Central connections of the sensory and motor nuclei of the vagus nerve

Recent morphological and immunohistochemical studies bearing on the central pathways involved in processing vagal afferent information and in modulating the activity of vagal preganglionic neurons are summarized. The nucleus of the solitary tract (NTS), the principal recipient of first order vagal afferent inputs, projects to preganglionic cell groups of both divisions of the autonomic nervous system, to motor nuclei of cranial nerves that supply the face and tongue, to a series of 'relay' nuclei in the brainstem, and to a number of cell groups in the hypothalamus and the limbic region of the telencephalon that integrate autonomic, neuroendocrine and regulatory behavioral responses. With the exception of the cranial nerve motor nuclei, each cell group in receipt of direct inputs from the NTS projects back to this region and/or to the vagal motor nuclei, and is thereby in a position to influence vagal motor outflow. This central vagal system is further characterized by the presence of neurons that contain an impressive diversity of neuropeptides and monoamines. Examples are cited to illustrate how biochemically specified projections within this system are organized, and how they provide potential substrates for encoding information transfer between its components.     

Optical survey of neural circuit formation in the embryonic chick vagal pathway

The multiple-site optical recording technique with a voltage-sensitive dye, NK2761, was used to survey functional organization of neural circuits related to the vagus nerve in the embryonic chick brainstem. When we stimulated the vagus nerve, in addition to the responses in the vagal sensory nucleus (nucleus of the tractus solitarius (NTS)) and motor nucleus (dorsal motor nucleus of the vagus nerve (DMNV)) on the stimulated side, another response area was discriminated at the level of the pons/rostral medulla on the contralateral side. Characteristics of the contralateral optical signals suggested that they correspond to the neural activity in the second/higher-ordered nucleus of the vagal pathway, possibly the parabrachial nucleus, which receives inputs from the NTS. Blockade of non-N-methyl-d-aspartate (NMDA) receptors abolished the responses on the contralateral side, together with the postsynaptic firing in the NTS, suggesting the significance of non-NMDA receptor function in sensory information transfer via the NTS. The responses on the contralateral side were first detected from the 7-day-old embryonic stage, when the glutamatergic excitatory postsynaptic potentials were first expressed in the NTS. The results suggest that the synaptic pathway from the NTS to the contralateral nucleus is already generated when the primary vagal afferents make functional synapses on NTS neurons.     

The central neural connections of the area postrema of the rat

We applied the neuroanatomical tracers cholera toxin-horseradish peroxidase and wheat germ agglutinin-horseradish peroxidase to investigate the neural connections of the area postrema (AP) in the rat. We find that the AP projects to the nucleus of the solitary tract (NTS) and dorsal motor nucleus of the vagus bilaterally both rostral and caudal to obex; the nucleus ambiguus; the dorsal aspect of the spinal trigeminal tract and nucelus and the paratrigeminal nucleus; the region of the ventrolateral medullary catecholaminergic column; the cerebellar vermis; and a cluster of structures in the dorsolateral pons which prominently include a discrete set of subnuclei in the lateral parabrachial nucleus. The major central afferent input to the area postrema is provided by a group of neurons in the paraventricular and dorsomedial hypothalamic nuclei whose collective dendrites describe a horizontally oriented plexus which encircles the parvocellular nucleus of the hypothalamus bilaterally. In addition, the caudal NTS may project lightly to the AP. The lateral parabrachial nucleus provides a very light input as well. These connections, when considered in the context of the known vagal afferent input and reduced blood-brain barrier of AP, place this structure in a unique position to receive and modulate ascending interoceptive information and to influence autonomic outflow as well.     

Neural control of blood glucose homeostasis; Effect of microinjection of glucose into hypothalamic nuclei on efferent activity of pancreatic branch of vagus nerve in the rat

Efferent discharges were recorded from the nerve filament dissected from the central cut end of the pancreatic branch of the vagus nerve in the rat. Microinjections of 5% glucose solution (100–200 nl) into the LHA caused an increase in efferent activity, however, those into VMH caused no change in discharge rate. The results of experiments indicate that activation of vagal pancreatic efferents in hyperglycemie situation is originated in LHA and transmitted to the vagal pancreatic motoneurons. The role played by the neural network on blood glucose homeostasis was also discussed.     

Development of obesity and neurochemical backing in aurothioglucose-treated mice

To clarify the neurochemical backing of aurothioglucose (ATG)-induced obesity in mice, we investigated lesion sites, hypothalamic neurotransmitters and c-Fos-like immunoreactivity (Fos-IR). At day 2 after ATG, tissue loss or cells death was observed in several parts of the ventral area of the ventromedial hypothalamic nucleus (VMH), and the dorsal area of arcuate nucleus and in the nucleus of the solitary tract (NTS). However, the greater part of the VMH was retained. Body weight began to increase in week 1. Hypothalamic serotonin (5-HT) and the metabolites were increased at day 2. The contents of acetylcholine, norepinephrine and dopamine in the hypothalamus showed no significant change. In week 1, the area shown tissue loss was compacted and plugged up. In the control group, most obvious c-Fos-like immunoreactive region was paraventricular nucleus (PVN). At day 2, Fos-IR was observed around destroyed regions in the hypothalamus and NTS, but few Fos-IR was found in the other regions including PVN. The Fos-IR around destroyed regions diminished after week 1. In week 3, Fos-IR in the PVN increased. These results suggest that the development of ATG-induced obesity cannot be attributed to solely VMH destruction. The restoration processes of the neuronal dysfunction involving PVN seem to play an important role in the development of obesity. NTS lesion and 5-HT system might contribute to decrease in food intake for several days after ATG.     

Phaseolus vulgaris leuco-agglutinin tracing of intrahypothalamic connections of the lateral, ventromedial, dorsomedial and paraventricular hypothalamic nuclei in the rat

Intrahypothalamic connections of the lateral (LHA), ventromedial (VMH), dorsomedial (DMH) and paraventricular (PVN) hypothalamic nuclei were studied with anterograde transport of iontophoretically injected Phaseolus vulgaris leuco-agglutinin and the immunocytochemical detection of labeled structures. The LHA was found to give rise to a minor projection in the VMH, whereas the VMH in reverse maintains few connections with the ventromedial part of the tuberal LHA. Tracer deposits in both the LHA and VMH resulted in anterograde terminal labeling in the DMH. The DMH, in turn, donates a small number of projections to the LHA and VMH. The main projection of the DMH is aimed at the parvocellular paraventricular nucleus. Direct outflow pathways from the VMH to the PVN were not found, but lectin injections in the LHA on the other hand gave rise to terminal labeling in both the parvocellular and magnocellular divisions of the PVN. The PVN in turn was found to give only minor reciprocal projections to the LHA, DMH and VMH. These findings indicate that the main stream of connections in the hypothalamus runs from the LHA and VMH to the DMH, and from the DMH to the PVN. The identified circuitry patterns were discussed with respect to the role of the hypothalamus in the control of homeostasis and metabolic regulation, and more specifically in relation to the modulation of the hormone release from the pancreas and adrenal glands.     

Chemical coding of the hypothalamic neurones in metabolic control. II. Norepinephrine-sensitive neurones and glycogen breakdown in liver

Our previous study has demonstrated that acetylcholine-sensitive neurones of the lateral hypothalamic nucleus are specifically concerned in the regulation of liver glycogen synthesis. The present study was undertaken to specify the ventromedial hypothalamic (VMH) neurones involved in regulation of liver glycogen breakdown, by means of chemical stimulation of the VMH with different neurotransmitters. Microinjection of norepinephrine (5 × 10⁻¹⁰−5 × 10⁻⁸ mol) into the VMH of rats caused a rapid increase in liver phosphorylase-a activity, the active form of this key glycogenolytic enzyme. Focal application of acetylcholine, serotonin, or GABA, on the other hand, did not affect appreciably the enzyme activity. Administration of dopamine into the VMH slightly decreased the phosphorylase-a activity.The effect of noradrenergic stimulation of the VMH on liver phosphorylase was blocked by previous intrahypothalamic application of propranolol, but was not by phentolamine. The effect of norepinephrine was also abolished by previous intraperitoneal injection of hexamethonium. These results indicate that, among other neurones, norepinephrine-sensitive neurones in the VMH are involved in regulation of phosphorylase activity in the liver and the effect of norepinephrine seems to be linked to β-adrenoreceptors in the hypothalamus. They also suggest that a noradrenergic component in the VMH-splanchnic nerve pathway is an important neural regulatory system for hepatic glycogen breakdown.     

Effects of Cholecystokinin in the Supraoptic Nucleus and Paraventricular Nucleus are Negatively Modulated by Leptin in 24‐h Fasted Lean Male Rats

Cholecystokinin (CCK) and leptin are two important satiety factors that are considered to act in synergy to reduce meal size. Peripheral injection of CCK activates neurones in several hypothalamic nuclei, including the supraoptic (SON) and paraventricular (PVN) nuclei and neurones in the brainstem of fed rats. We investigated whether peripheral leptin would modulate the effects of CCK on neuronal activity in the hypothalamus and brainstem of fasted rats by investigating Fos expression in the PVN, SON, arcuate nucleus, ventromedial hypothalamus (VMH), dorsomedial hypothalamus (DMH), area postrema (AP) and the nucleus tractus solitarii (NTS). Male rats, fasted for 24 h, received either one i.p. injection of vehicle, leptin or CCK‐8 alone, or received one injection of vehicle or leptin before an i.p. injection of CCK‐8. We found that CCK increased Fos expression in the PVN and SON as well as in the NTS and AP, but had no effect on Fos expression in the arcuate nucleus, VMH or DMH compared to vehicle. Leptin injected alone significantly increased Fos expression in the arcuate nucleus but had no effect on Fos expression in the VMH, DMH, SON, PVN, AP or NTS compared to vehicle. Fos expression was significantly increased in the AP in rats injected with both leptin and CCK compared to rats injected with vehicle and CCK. Unexpectedly, there was significantly less Fos expression in the PVN and SON of fasted rats injected with leptin and CCK than in rats injected with vehicle and CCK, suggesting that leptin attenuated CCK‐induced Fos expression in the SON and PVN. However, Fos expression in the NTS was similar in fasted rats injected with vehicle and CCK or with leptin and CCK. Taken together, these results suggest that leptin dampens the effects of CCK on Fos expression in the SON and PVN, independently from NTS pathways, and this may reflect a direct action on magnocellular neurones.     

Modulation of lateral hypothalamic activity by olfactory bulb and sciatic stimulation.

Lateral hypothalamic area (LHA) single unit activity (extracellular) was studied in response to electrical stimulation of the olfactory bulb (OB) or sciatic nerve in adult albino rats (n = 39) anesthetized with dialurethane. Olfactory stimulation resulted in a greater proportion of LHA units showing inhibitory rather than excitatory responses, while sciatic nerve stimulation resulted in similar proportions of units showing inhibitory and excitatory responses. Of the 76 LHA units tested with both OB and sciatic nerve stimulation, 36% responded to both stimulation sites, 18% responded only to OB stimulation, 26% responded only to sciatic nerve stimulation, and 20% were unresponsive to either stimulation. The locations of responsive units were diffuse throughout the LHA sampled. The response characteristics of LHA neurons to external sensory stimulation are consistent with the anatomy and putative integrative functions of this brain region.     

Effects of cholecystokinin on sympathetic activity to interscapular brown adipose tissue.

The effects of injecting cholecystokinin (CCK) into the third ventricle or into selected hypothalamic sites on electrical firing rate of sympathetic nerves to interscapular brown fat (IBAT) has been investigated in anesthetized rats. The hypothesis for these experiments was that there was a reciprocal relationship between sympathetic activity and food intake. Since CCK reduces food intake we predicted that CCK would stimulate sympathetic activity to IBAT. Following the injection of CCK into the third ventricle there was an increase in firing rate of sympathetic nerves to IBAT. When the peptide was injected into either the ventromedial hypothalamic nucleus (VMH) or lateral hypothalamic area (LHA), there was likewise an increase in sympathetic firing rate. The injection of CCK into the paraventricular nucleus produced a small decrease in sympathetic firing rate. In contrast, no effect was seen following injection of CCK into the preoptic area or dorsomedial hypothalamic nucleus. Thus, the VMH or LHA appear to be the principal hypothalamic areas mediating the stimulation of sympathetic activity to IBAT which is observed following the third ventricular injection of CCK. These studies support the hypothesis of a reciprocal relationship between the effects of CCK on the thermogenic component of the sympathetic nervous system and food intake and identify the VMH and LHA as the primary sites for this effect.     

Effects of lateral hypothalamic stimulation on medulla oblongata and gastric vagal neural responses

Recordings were made of neural activity in the medial to lateral region of the dorsal nucleus of the vagus in the medulla oblongata (NDV), and from the gastric branch of the vagal nerve (gastric vagus) in rats. Gastric acid secretion following lateral hypothalamic (LHA) stimulation was observed, and NDV neurons were identified by stimulation of the peripheral end of the gastric vagus. NDV-neurons responded to LHA stimulation with latencies of about 5 msec, and about 6.5 msec to the peripheral stimulation of the gastric vagus. Out of 274 NDV neurons, which were located by their spontaneous discharge, 186 (67.9%) responded to LHA stimulation. Gastric acid secretion (with either short or long latency) occurred in 8.6% (16/186) of these cases. These 16 neurons were considered to be ‘gastric secretory’ neurons and are discussed as such. The results imply that some LHA neurons, which are either concerned with or directly control gastric acid secretion, communicated by at least one path (probably polysynaptic) to the medulla oblongata and then via the vagus to the oxyntic cells of gastric glands.