Importance of endogenous nitric oxide synthase in the rat hypothalamus and amygdala in mediating the response to capsaicin.

Although capsaicin has been shown to activate certain neuronal groups in the hypothalamus and amygdala, the neurotransmitters involved and the exact mechanism of action are not clearly understood at present. The aim of this study was to examine the hypothesis that the effect of capsaicin in the rat hypothalamus and amygdala primarily involves direct activation of the endogenous nitric oxide synthase (NOS) neurons responsible for the synthesis of nitric oxide (NO). Subcutaneous capsaicin injection in male rats, compared with vehicle, caused a significant increase in Fos expression in the paraventricular nucleus (PVN), supraoptic nucleus (SON), and medial and cortical amygdala. The expression of nicotinamide adenine dinucleotide phosphate diaphorase, a histochemical marker for NOS, was also increased in these brain areas in addition to the periventricular and lateral hypothalamic area and central amygdaloid nucleus. Also, capsaicin significantly increased the expression of neuronal NOS messenger RNA and protein in the PVN, SON, and medial amygdala as demonstrated by in situ hybridization and immunohistochemistry, respectively. A higher proportion of the NOS neurons in the PVN, periventricular region, SON and amygdala showed Fos expression in response to capsaicin than vehicle injection. There was little, if any, Fos activation in the NOS-positive neurons in the lateral hypothalamic area. The capsaicin-induced activation of the hypothalamic PVN and SON neurons and the medial amygdaloid nucleus was attenuated in the NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) -pretreated animals in comparison with the inactive enantiomer D-NAME. These observations indicate that activation of the endogenous NOS system and production of NO constitute a major pathway through which capsaicin exerts its effect within the hypothalamus and amygdala.

Gabaergic control of olfactory learning in young rats.

Olfactory learning in young rats correlates with neural plasticity in the olfactory bulb, and involves noradrenergic modulation of reciprocal dendrodendritic synapses between mitral cells and GABAergic granule cells. The purpose of this study was to examine, in vivo, the consequences of manipulating bulbar GABA transmission during training. In the first experiment, postnatal day 11 rat pups were trained in an olfactory associative learning task with citral odor and foot shock as the conditioned and unconditioned stimuli, respectively. The pups received continuous infusion of saline or the GABA(A) receptor agonist muscimol into the olfactory bulbs throughout a 30-min training session. The pups were then tested on postnatal day 12 for a preference for or an aversion to citral odor. Saline-infused control pups developed an aversion to citral odor. The GABA(A) receptor agonist muscimol impaired this aversive learning in a dose-dependent manner. In the second experiment, pups were exposed to the odor for 30 min while receiving continuous intrabulbar infusion of a low or high dose of the GABA(A) receptor antagonist bicuculline, without any other reinforcer. Depending on whether a low (0.2 nmol/bulb) or high (1.0 nmol/bulb) dose of bicuculline was infused, the pups showed a preference or an aversion for citral odor after infusion of low and high doses, respectively. These results indicate that disinhibition of mitral cells in the olfactory bulb is critical for olfactory learning in young rats, and suggest that the degree of disinhibition is an important determinant in acquiring either preference or aversion for the conditioned odor.

A role for nitric oxide in the median eminence and arcuate nucleus response to capsaicin treatment in rats.

This study examined a possible functional involvement of nitric oxide (NO) in the median eminence (ME) and arcuate nucleus (ARC) after capsaicin treatment in rats. Subcutaneous injection of capsaicin increased nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the ARC-ME compared with vehicle treatment. Fos expression was increased in the ARC after capsaicin injection compared with vehicle-treated rats. Pretreatment with the NO synthase (NOS) inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME) attenuated the effect of capsaicin on Fos expression and NADPH-d reactivity in the ARC-ME in comparison with rats injected with D-NAME, the inactive stereoisomer of L-NAME. These observations suggest that NO makes a major contribution to the response of the ARC-ME to a stressor such as capsaicin.

The role of limbic-hypothalamic systems in metabolic responses of phenylalanine to repeated cold exposures in rabbits.

In this study we examine the effects of lesions to limbic or hypothalamic structures on the metabolic responses of phenylalanine in liver slices prepared from rabbits exposed to cold stress (-20 degrees C) for 12 hours at a fixed time once a day. The results were as follows: 1. The 1st cold exposure (cold exposure on the 1st day) had marked and various effects on phenylalanine metabolism, and the metabolic responses of phenylalanine to cold exposure gradually decreased and then completely disappeared with repetitions of exposure in intact rabbits. 2. The metabolic patterns and responses of phenylalanine to the 1st cold exposure were altered by lesioning the periventricular arcuate nucleus (ARC), ventromedial hypothalamus (VMH), stria terminalis (ST) or dorsal fornix (FX). 3. The complete abolishment of the metabolic responses to cold exposure by its repetition was observed in rabbits with lesions in the ARC or VMH as well. 4. In contrast, metabolic responses of phenylalanine to cold exposure in rabbits with lesions in the ST or FX remained even after six exposures. These results suggest that the ARC, VMH, amygdala (AMYG)-ST system and dorsal hippocampus (HPC)-FX system played a role in the metabolic regulation of phenylalanine and in the metabolic responses of phenylalanine to the 1st cold exposure and that the AMYG-ST and HPC-FX systems, but not the ARC or VMH, participated in the process of the metabolic adaptation of phenylalanine to cold exposure.

Influence of the electrical stimulation of the hypothalamus on adrenocortical steroidogenesis in hypophysectomized rats.

The electrical stimulation of the ventromedial hypothalamus (VMH), lateral hypothalamic area (LHA), periventricular arcuate nucleus (ARC), and posterior hypothalamus (PHY), on 14C transfer rates from 14C-1-acetate into adrenocortical steroids in adrenal slices of hypophysectomized rats were investigated. The 14C transfer rates into corticosterone and cortisol were increased by the stimulation of the VMH, ARC, and PHY, but decreased by the stimulation of the LHA. From these results, it might be suggested that these hypothalamic structures were involved in the regulation of adrenocortical steroidogenesis without participation of the pituitary.

Olfactory recognition: a simple memory system.

Mice have an olfactory (pheromone) recognition memory located at the first relay in the sensory system. It is acquired with one-trial learning, contingent upon norepinephrine activation at mating, and lasts for several weeks. The mechanism involves Hebbian (association-dependent) changes in synaptic efficacy at dendrodendritic synapses in the accessory olfactory bulb. As a result of this memory, males made familiar by mating are recognized by the females, thereby mitigating pregnancy block. Such a memory function is biologically important to the female, as it is required to sustain pregnancy in the presence of her stud male's odors.

Influence of electrical stimulation of the hypothalamus on ovarian steroidogenesis in hypophysectomized and adrenalectomized rats.

The effects of electrical stimulation of the lateral hypothalamic area (LHA), periventricular arcuate nucleus (ARC) and ventromedial hypothalamic nucleus (VMH) on the rates of 14C transfer from 14C-1 acetate into ovarian steroids in ovarian slices of hypophysectomized and adrenalectomized (H-A) rats were investigated. The 14C transfer rates into estrogen and 20 alpha-hydroxypregn-4-en-3-one (20 alpha-OH-P) were decreased by LHA stimulation. The stimulation of the ARC and VMH increased the rates of 14C transfer into estrogen, progesterone and 20 alpha-OH-P. From these results, it might be suggested that these hypothalamic structures were involved in the regulation of ovarian steroidogenesis without participation of the pituitary and adrenal.

Influence of lesions of the limbic-hypothalamic system on metabolic response of pyruvate to daily repeated immobilization stress in rabbits.

The pyruvate metabolic response to the 1st exposure (exposure on the 1st day) to immobilization stress (IMO) were considerably altered by lesions of the periventricular arcuate nucleus (ARC), ventromedial hypothalamus (VMH), stria terminalis (ST) and dorsal fornix (FX). The pyruvate metabolic responses to IMO were completely abolished by seven times repetition of exposure to IMO in the rabbits with lesions of ARC and VMH; they were similar to sham-operated groups. In rabbits with lesions of ST and FX, the pyruvate metabolic responses to the 7th exposure (exposure on the 7th day) to IMO were almost the same as those after the 1st exposure to IMO, but these metabolic responses were completely abolished by the seven times repetition of exposure to IMO in the sham-operated animals. These results suggest that firstly the ARC, VMH, amygdala (AMYG)-ST system and dorsal hippocampus (HPC)-FX system are involved in the pyruvate metabolic responses to the 1st exposure to IMO, and secondly, that the AMYG-ST system and the HPC-FX system are involved in the disappearance process of pyruvate metabolic responses to IMO by the daily repetition of exposure to IMO.

Subfornical organ efferents influence the activity of median preoptic neurons projecting to the hypothalamic paraventricular nucleus in the rat.

Twelve neurons in the median preoptic nucleus were antidromically activated by electrical stimulation of the hypothalamic paraventricular nucleus in the rat. Electrical stimulation of the subfornical organ produced excitation (N = 7) or inhibition (N = 4) of the activity of these identified units. The activity of all identified units that were excited by stimulation of the subfornical organ was also excited by microiontophoretically applied angiotensin II whereas the remaining units were not affected. The excitatory responses of the identified units to subfornical organ stimulation or applied angiotension II were blocked by saralasin, an angiotensin II antagonist.

Ventrolateral medullary neurons projecting to the medial preoptic/anterior hypothalamic area through the medial forebrain bundle: an electrophysiological study in the rat.

A total of 152 ventrolateral medullary neurons was antidromically stimulated from both the medial preoptic/anterior hypothalamic area (MPOAH) and the medial forebrain bundle (MFB) in urethane anesthetized rats. These neurons were located primarily dorsal to the lateral reticular nucleus and could be readily classified in at least two groups, type I and type II cells on the basis of electrophysiological properties. The action potentials of type I cells had a shorter duration, and their conduction velocities ranged from 0.45 to 3.1 m/s. By contrast, type II cells, most predominantly observed, were characterized by a longer duration and an unusual shape of their action potential, and the antidromic propagation into the somatodendritic complex was often blocked. The conduction velocity (mean = 0.21 m/s) and absolute refractory period (mean = 2.63 ms) of type II cells are consistent with them having fine non-myelinated axons. Injection of 6-hydroxydopamine (6-OHDA), but not 5,7-dihydroxytryptamine, directly into the MFB blocked antidromic responses of 57% of type II cells tested. The residual type II cells whose antidromic responses were not affected by 6-OHDA were located significantly rostral to the 6-OHDA sensitive cells. Neither antidromic response of type I cells tested, on the other hand, was affected by 6-OHDA. The majority of type I cells were dramatically activated by noxious pinches of the tail, whereas the noxious stimuli produced no detectable change in the firing of type II cells. These data demonstrate that ventrolateral medullary neurons projecting to the MPOAH through the MFB are comprised of at least three distinct populations: 6-OHDA resistant fast conducting cells with somatic afferents, 6-OHDA sensitive and resistant slow conducting cells.

Influence of lesions of the limbic-hypothalamic system on adrenocortical responses to daily repeated heat exposures in rabbits.

The effects of lesions in the basal medial hypothalamus and limbic structure upon the responses of adrenocorticoids formation in adrenal slices of rabbits to daily repeated heat exposures has been investigated. (1) The adrenocortical responses to heat exposure on the 1st day were decreased by lesions in the periventricular arcuate nucleus (ARC), ventromedial hypothalamus (VMH), stria terminalis (ST) and dorsal fornix (FX). (2) There were no effects of heat exposure on the 10th day upon the adrenocorticoid formation in either the sham-lesioned rabbits or the rabbits with the lesions of ARC, VMH and ST. (3) In rabbits with the FX lesions, the adrenocorticoids formation was significantly increased by heat exposure on the 10th day. (4) These results suggested that the basal medial hypothalamus, amygdala (AMYG)-ST system and dorsal hippocampus (HPC)-FX system participated in the mechanisms of adrenocortical responses to heat exposure on the 1st day, but only the HPC-FX system played some roles in complete disappearance process of adrenocortical responses to heat exposure by repetition of exposures.

Influence of lesions in the limbic-hypothalamic system on adrenocortical responses to daily repeated exposures to immobilization stress in rabbits.

In order to elucidate the role of the limbic-hypothalamic system in the adrenocortical responses to repeated immobilization stress, the influences of daily repeated exposures of immobilization stress on the adrenocorticoid formation in adrenal slices of rabbits with lesions of various sites of the hypothalamus and limbic structures had been investigated. The experimental results obtained were as follows: (1) Lesions in periventricular arcuate nucleus (ARC) and ventromedial hypothalamus (VMH) decreased the rates of adrenocortical responses to the 1st immobilization exposure (exposure on the 1st day), but did not alter the rates of adrenocortical responses to the 4th immobilization exposure (exposure on the 4th day). (2) Lesions in stria-terminalis (ST) and dorsal fornix (FX) did not alter the rates of adrenocortical responses to the 1st immobilization exposure, but increased the rates of adrenocortical responses to the 4th immobilization exposure. (3) The adrenocortical responses to immobilization stress were completely abolished by the seven times repetition of exposures in each sham-operated group and in the animals with lesions of ARC and VMH, but in the rabbits with lesions of ST and FX the adrenocortical responses to immobilization stress were not abolished by the seven times repetition of exposures. (4) These results suggested that such basal medial hypothalamus participated in the mechanisms of adrenocortical responses to the 1st exposure of immobilization stress, but did not participate in the disappearance process of adrenocortical responses to immobilization stress by the seven times repetition of exposures. On the other hand, it might be supposed that the amygdala(AMYG)-ST system and dorsal hippocampus(HPC)-FX system do not participate in the mechanism of adrenocortical responses to the 1st immobilization exposure, but these limbic areas played some roles in the disappearance process of the adrenocortical responses to immobilization stress by the seven times repetition of exposures.

Antidromic identification of neurons in the ventrolateral part of the medulla oblongata with ascending projections to the preoptic and anterior hypothalamic area (POA/AHA).

Seventy neurons in the ventrolateral medulla oblongata were antidromically activated by electrical stimulation of the preoptic and anterior hypothalamic area (POA/AHA) in female rats under urethane anesthesia. These identified cells were located within and adjacent to the nucleus reticularis lateralis and could be readily distinguished into at least two types of neurons, designated as "fast' and "slow' cells, on the basis of their waveform and conduction velocity.

[Nutritional evaluation of protein concentrates of beans (Phaseolus vulgaris) and lentils (Lens esculenta)]. / Evaluación nutricional de concentrados proteicos de porotos (Phaseolus vulgaris) y de lentejas (Lens esculenta).

The composition and nutritive value were determined in navy bean meal (Phaseolus vulgaris) and lentil meal (Lens esculenta), and in their respective protein concentrates obtained through extraction followed by isoelectric precipitation. Sulfur amino acids per gram of nitrogen were lower in the concentrates than in the meals, while there was no difference for lysine and threonine. The white bean protein concentrate had a lower biological value than the meal but better digestibility, although trypsin inhibitor concentration was unchanged. Digestibility greatly improved with heating but it did not increase beyond 81% even after autoclaving. Autoclaved samples supplemented with methionine reached a biological value of 83. The lentil protein concentrate also had a lower biological value than the meal but digestibility was high for both samples (91%) and remained unchanged after heating. Trypsin inhibitors were absent. After supplementing with methionine, a biological value of only 63 was obtained, due to the low level of tryptophan, the second limiting amino acid. In spite of the concentrates' lower biological value, it was proved that they equalled the meals' potential for complementing cereal, as their content in lysine and threonine is high. The concentrates have the additional advantage of allowing effective supplementation without increasing the legume-cereal ratio.