Release of classical transmitters and nitric oxide in the rat olfactory bulb, evoked by vaginocervical stimulation and potassium, varies with the oestrus cycle

In vivo microdialysis was used to investigate the effects of ovariectomy and the oestrus cycle on vaginocervical stimulation-evoked classical transmitter and nitric oxide release in the olfactory bulb of anaesthetized (urethane) and conscious rats. During pro-oestrus/oestrus, vaginocervical stimulation (1 or 10 min) significantly increased concentrations of glutamate, aspartate, GABA, noradrenaline, dopamine and nitric oxide (citrulline) but failed to do so in met-oestrus/di-oestrus or following ovariectomy. Potassium chloride-evoked GABA, noradrenaline and nitric oxide release in the olfactory bulb was also significantly enhanced during pro-oestrus/oestrus. The effects of vaginocervical stimulation on olfactory bulb transmitter release during pro-oestrus/oestrus were significantly reduced by pelvic or vagus nerve section. Basal concentrations of classical transmitters and nitric oxide in the olfactory bulb did not vary across the oestrus cycle although noradrenaline and dopamine levels were reduced following ovariectomy. These results confirm our previous electrophysiological data showing that the olfactory bulb mitral cells are only excited by vaginocervical stimulation during pro-oestrus/oestrus. They also suggest that sex hormones acting primarily at the level of the olfactory bulb dramatically enhance the ability of vaginocervical stimulation to evoke release of both classical transmitters and nitric oxide in this region. Such alterations in neurochemical release in the olfactory bulb may be important for mediating plasticity changes underlying olfactory recognition of mates or offspring.     

颞叶癫痫大鼠不同时期海马细胞外氨基酸类递质的变化

目的探讨海人酸(Kainic Acid,KA)诱发颞叶癫痫大鼠不同时期海马CA3区细胞外谷氨酸(gluta-mate,GLU)、γ-氨基丁酸(gamma-aminobutyric acid,GABA)、甘氨酸(glycine,GLY)、天冬氨酸(aspartic acid,ASP)、牛磺酸(taurine,TAU)等递质水平的变化。方法 21只SD大鼠随机分为对照组(9只)和KA组(12只)。KA组大鼠海马CA3区注射0.8μgKA,对照组海马CA3区注射同体积生理盐水,于1天(急性期)、7天(静止期)、30天(慢性期)对大鼠海马CA3区行微透析,用高效液相色谱法(high-performance liquid chromatography,HPLC)测定透析液中GLU、GLY、GABA、ASP、TAU含量的变化。结果急性期:与对照组相比,KA组CA3区细胞外GLU(P=0.004)、GLU/GABA(P=0.002)明显增高,GABA(P=0.001)、GLY(P<0.001)明显减少,ASP(P=0.23)、TAU(P=0.28)与对照组相比无明显差异。静止期:KA组GLU(P=0.003)、GLU/GABA(P<0.001)较对照组明显降低,而GABA(P<0.001)、ASP(P<0.001)、TAU(P<0.001)较对照组明显增高,GLY(P=0.49)与对照组相比无明显差异。慢性期:与对照组相比,KA组GLU(P<0.001)、GABA(P=0.013)、ASP(P<0.001)、TAU(P<0.001)、GLU/GABA(P<0.001)明显升高,GLY与对照组相比无明显差异(P=0.86)。结论海马CA3区兴奋性与抑制性氨基酸类递质失衡可能是癫痫发生发展的重要机制之一。     

NMDA and Kainate-evoked Release of Nitric Oxide and Classical Transmitters in the Rat Striatum: In Vivo Evidence that Nitric Oxide May Play a Neuroprotective Role

The effects of N-methy-d -aspartate (NMDA), kainate, S-α-amino-3-hydroxyd-5-methyl-4-isoxazole propionate (AMPA) and KCI on striatal nitric oxide (NO), acetylcholine (ACh), dopamine (DA), serotonin (5-HT), aspartate (ASP), glutamate (GLU) and γ-aminobutyric acid (GABA) release were measured in anaesthetized rats in vivo by microdialysis and in vitro in organotypic slice cultures. Local NMDA (1–100 μM) infusion by retrodialysis dose-dependently increased levels of classical transmitters, NO₂-, NO₃-, ctrulline and arginine at similar thresholds (10 γM) Similar patterns of NMDA-evoked (50 μM) release were seen in striatal cultures. NMDA-evoked changes were all calcium-dependent and blocked by NMDA (APV or MK-801) but not AMPN/kainate (DNQX) receptor antagonists, excepting DA which could be prevented by both. In vivo, kainate increased NO₂^-, NO₃^-, CIT and ARG levels at 50 and 100 μM but was less potent than NMDA. Kainate also evoked significant Ach₁ DA and GLU release dose-dependently starting at 1–10 μM whereas 5-HT, ASP and GABA required 50 or 100 μM doses. Kainate effects were inhibited by DNQX, but not by APV, and were calcium-dependent. AMPA failed to alter NO₂^-, NO₃^-, CIT or ARG levels at 50 or 100 μM doses but dose-dependently increased ACh and DA. Similar results were seen with kainate (50 μM) and AMPA (50 μM) in vitro KCI evoked NO₂^-, NO₃^-, CIT and ARG release as well as that of the classical transmitters in vivo and in vitro. In vivo administration of the NO synthase inhibitor L-nitroarginine (L-NARG; 100 μM) significantly reduced NO₂^-, NO₃^- and CIT levels and prevented NMDA, kainate or KCI-evoked increases. It also potentiated ACh, ASP, GLU and GABA release and reduced that of DA in response to 50 μM NMDA whereas treatment with an NO-donor (SNAP; 10 μM) significantly reduced evoked ACh, ASP and GLU release. The NO synthase inhibitor L-NARG potentiated kainate-evoked ACh release and reduced that of DA, although less potently than NMDA, but it had no effect on KCI-evoked transmitter release. Overall, these results show that both NMDA and kainate increase striatal NO release at similar dose-thresholds as for classical transmitter release suggesting that NO is dynamically released under physiological and not just pathological conditions. Reduction of striatal NO levels also potentiates calcium-dependent transmitter release in response to NMDA and, to a lesser extent, kainate, whereas increasing them reduces it. This is consistent with a role for NO as a neuroprotective agent in this region acting to desensitize NMDA receptors.     

Role of Nitric Oxide in Modulating the Release of Dopamine, Glutamate, and GABA in Striatum of the Freely Moving Rat

This study investigated the role of nitric oxide (NO) in modulating the basal and N-methyl-

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-aspartate (NMDA)-induced release of dopamine (DA), glutamate (GLU), and γ-aminobutiric acid (GABA) in striatum of the freely moving rat using microdialysis. Intrastriatal infusion of NMDA (5 mM) for 15 min increased extracellular concentrations of DA, GLU, and GABA. NMDA also decreased extracellular concentrations of DA metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC), and 4-hydroxy-3-methoxyphenylacetic acid (HVA), and of the GLU and GABA precursor, glutamine (GLN). Perfusion of N-nitroarginine (1–5 mM), an inhibitor of the synthesis of NO, potentiated NMDA-induced increases in extracellular concentrations of DA and attenuated increases of extracellular GLU. NMDA-induced decreases of extracellular concentrations of DOPAC were also attenuated by N-nitroarginine. N-nitroarginine had no effect on NMDA-induced changes of extracellular concentrations of GABA, HVA, and GLN. N-nitroarginine decreased basal concentrations of DOPAC and HVA, and increase basal concentrations of GLN, but had no effect on basal DA, GLU, and GABA. These results suggest a role for NO in modulating the NMDA-induced release of DA and GLU in striatum. They also suggest that NO could be regulating the basal metabolism of DA, GLU, and GABA.     

Activation of arginine metabolism to glutamate in rat brain synaptosomes in thioacetamide-induced hepatic encephalopathy: An adaptative response?

Crude (P2) synaptosomes derived from rats with acute hepatic encephalopathy (HE) induced with thioacetamide showed a slightly increased uptake of radiolabeled arginine (ARG) and a 2.5-fold enhanced conversion of newly taken-up ARG to both glutamate (GLU) and gamma-aminobutyric acid (GABA) as compared with control synaptosomes. Pulse treatment of the preloaded synaptosomes with a high potassium medium decreased their radioactive GLU and GABA content without affecting the content of the precursor ARG. This result, which was identical with control or HE preparations, appears to indicate that ARG contributes at least, in part, to the synthesis of neurotransmitter GLU or GABA. As measured in purified synaptosomal preparations, HE increased by about 50% the activities of arginase and ornithine-delta-aminotransferase--the two enzymes of the ARG to GLU shunt. It is postulated that increased conversion of ARG to GLU may compensate for excessive utilization of the latter amino acid as an ammonia trap during HE and, as such, may be considered as an adaptative response of the synaptic compartment to this pathological condition.     

Intracerebral microdialysis of extracellular amino acids in the human epileptic focus.

Extracellular levels of aspartate (ASP), glutamate (GLU), serine (SER), asparagine (ASN), glycine (GLY), threonine (THR), arginine (ARG), alanine (ALA), taurine (TAU), tyrosine (TYR), phenylalanine (PHE), isoleucine (ILEU), and leucine (LEU) were monitored by using intracerebral microdialysis in seven patients with medically intractable epilepsy, undergoing epilepsy surgery. In association with focal seizures, dramatic increases of the extracellular ASP, GLU, GLY, and SER concentrations were observed. The other amino acids analyzed, including TAU, showed small changes. The results support the hypothesis that ASP, GLU, GLY, and possibly SER, play an important role in the mechanism of seizure activity and seizure-related brain damage in the human epileptic focus.     

Neurochemical monitoring in the management of severe head-injured patients with hypothermia

The neuroprotective action and effect of hypothermia on the neurochemical system is not well understood. The present study was performed using six patients with GCS scores of 5 or less to clarify the relationship between monitored brain temperature, intracranial pressure (ICP), cerebral perfusion pressure (CPP) and oxygen saturation of the jugular venous blood (SjO2). Changes in concentration of excitatory amino acids, glutamate (GLU) and aspartate (ASP), and NO2 were studied using intracerebral microdialysis as well as in jugular venous blood and cerebrospinal fluid (CSF). Changes in brain temperature, CPP and SjO2 resulting from hypothermia and brain death associated with markedly higher concentrations of and fluctuations in the concentrations of GLU, ASP and NO2 were observed in the dialysate than in the jugular venous blood or CSF. Hypothermic treatment significantly reduces excitatory amino acid and NO2 concentrations, a finding which was associated with an improvement in CPP and SjO2. Measurement of GLU and ASP using intracerebral microdialysis is a clinically useful method for clarifying abnormal neurochemical events associated with severe head injury and for evaluating the effects of hypothermia.     

Microdialysis measurement of neurochemical changes in the mediobasal hypothalamus of ovariectomized ewes during oestrus

Oestrus behaviour and the luteinizing hormone (LH) surge are induced in ovariectomized ewes by oestradiol (E2) after a period of progesterone priming with a low level of E2 (Pge2) and we have previously shown that these effects are primarily mediated through their action on the mediobasal hypothalamus (MBH). The aim of the present study was to assess what neurochemical changes in the MBH are induced by these steroids that might mediate their action on oestrus behaviour and LH release. Eight ovariectomized ewes were implanted with microdialysis probes in the MBH and submitted to three artificial cycles, so that they exhibited either both oestrus behaviour and an LH surge (Pge2 + E2), and LH surge alone (E2 alone) or neither oestrus behaviour nor an LH surge (Pge2 alone). Microdialysis and blood samples were collected every 30 min from 4 h before the end of Pge2 treatment until the end of oestrus. Behavioural tests with a ram were made to assess receptivity. Dopamine (DA) levels were found to increase significantly at the termination of Pge2 treatment after both Pge2 + E2 and Pge2 treatments. When the ewes received E2 after a Pge2 + low estradiol priming (Pge2 + E2), DA levels decreased 16 h later (4 h after E2) whereas they did not change after E2 or Pge2 alone. By contrast, serotonin (5HT) levels did not change significantly during the first 24 h but then increased when ewes received E2 alone and decreased when they were treated with Pge2 + E2. γ-Aminobutyric acid (GABA) concentrations decreased significantly at the beginning of the sampling period after all treatments but this decrease lasted longer after Pge2 + E2 and was most pronounced at the beginning of receptivity. No significant long term effects of these steroid treatments were found on noradrenaline (NA), aspartate, glutamate, glycine and taurine levels. However, E2 administration was followed during the next few hours by a significant increase in glycine and to a smaller extent in glutamate and GABA. More importantly, when ewes were treated with Pge2 + E2, NA levels increased signicantly following the behavioural interactions with a ram when the ewes were sexually receptive. In contrast to this, DA levels only increased during interactions with the ram when the ewes were not receptive. 5HT levels increased after tests where the ewe was either receptive or unreceptive to the male. GABA, aspartate and glycine levels increased in the sample just preceding the test and then decreased during it. These results show that a number of neurotransmitter changes occur in the MBH during hormonal induction of oestrus. Changes mainly occured in DA, NA, 5HT and GABA concentrations during oestrus when animals were treated with Pge2 + E2 and suggest that they may be potentially involved in the control of female sexual behaviour. The changes occuring when animals received E2 alone and showed only prolactin and LH surges, but no behavioural oestrus, are much more limited. Furthermore, they do not correspond to changes after the E2 injection in the Pge2 + E2 treatment, so it is unlikely that the neurochemical changes we observed are directly related to these endocrine events.     

Prenatal ontogeny of substance P-like immunoreactivity in the nucleus tractus solitarii and dorsal motor nucleus of the vagus nerve of the human fetus: an immunocytochemical study

By using immunocytochemical method, the prenatal ontogeny of substance P-like immunoreactivity (SP-LI) was demonstrated in the dorsal motor nucleus of the vagus nerve (nX) and the nucleus tractus solitarii (nTS) of the human fetus at fetal age (menstruation age) of 11.5 weeks to 40 weeks. The time of initial appearance of SP-LI in the human brainstem nTS was between the fetal age 11.5 weeks and 16 weeks. At fetal age 16 weeks, the nTS showed moderate density of SP-LI fibers and terminals in subnucleus dorsalis of the nTS and nX. While the fetus grew, the density of SP-LI in the human fetus brainstem nTS and nX increased gradually from fetal age 16 weeks to 40 weeks. According to the Nissl staining, at fetal age 23 weeks, the nTS of human fetus can be subdivided into dorsal, medial, dorsolateral, ventrolateral and ventral gelatinosus subnuclei. The cytoarchitectonic subdivisions of human fetus nTS is in good agreement with the results obtained by immunocytochemical staining. These findings indicated that substance P (SP) might play an important role in the development of human brainstem nX, nTS, their related cranial nerves, and in their functional establishment during the pranatal period.     

Immunocytochemical organization and sour taste activation in the rostral nucleus of the solitary tract of mice

Sensory inputs from the oropharynx terminate in both the trigeminal brainstem complex and the rostral part of the nucleus of the solitary tract (nTS). Taste information is conveyed via the facial and glossopharyngeal nerves, while general mucosal innervation is carried by the trigeminal and glossopharyngeal nerves. In contrast, the caudal nTS receives general visceral information largely from the vagus nerve. Although the caudal nTS shows clear morphological and molecularly delimited subdivisions, the rostral part does not. Thus, linking taste‐induced patterns of activity to morphological subdivisions in the nTS is challenging. To test whether molecularly defined features of the rostral nTS correlate with patterns of taste‐induced activity, we combined immunohistochemistry for markers of various visceral afferent and efferent systems with c‐Fos–based activity maps generated by stimulation with a sour tastant, 30 mM citric acid. We further dissociated taste‐related activity from activity arising from acid‐sensitive general mucosal innervation by comparing acid‐evoked c‐Fos in wild‐type and “taste blind” P2X₂/P2X₃ double knockout (P2X‐dbl KO) mice. In wild‐type mice, citric acid stimulation evoked significant c‐Fos activation in the central part of the rostral nTS—activity that was largely absent in the P2X‐dbl KO mice. P2X‐dbl KO mice, like wild‐type mice, did exhibit acid‐induced c‐Fos activity in the dorsomedial trigeminal brainstem nucleus situated laterally adjacent to the rostral nTS. This dorsomedial nucleus also showed substantial innervation by trigeminal nerve fibers immunoreactive for calcitonin gene‐related peptide (CGRP), a marker for polymodal nociceptors, suggesting that trigeminal general mucosal innervation carries information about acids in the oral cavity. J. Comp. Neurol. 525:271–290, 2017. © 2016 Wiley Periodicals, Inc.     

Monoamine, amino acid and cholinergic interactions in slices of rat cerebral cortex

Interactions of monoamine, amino acid and cholinergic transmitter systems were studied in slices of rat cerebral cortex using a superfusion procedure and measuring release of endogenous dopamine (DA), norepinephrine (NE), serotonin (5-HT), GABA, glutamate (GLU) and aspartate (ASP). Depolarizing concentrations of K+ were used to induce a Ca2+-dependent, Mg2+-inhibited release of the monoamines and amino acids. Submaximal release of the monoamines and amino acids was observed at 35 mM K+, which permitted studies of possible excitatory or inhibitory actions of the added agents. The 35 mM K+-stimulated, Ca2+-dependent release of GABA was inhibited 40, 30 and 25% by 100 microM NE, DA and 5-HT, respectively. The release of GLU was potentiated by NE and reduced by DA. Both DA and 5-HT inhibited the release of ASP. The Ca2+-dependent, K+-stimulated release of endogenous NE, DA and 5-HT was not altered by 100 microM GABA, GLU or ASP. However, 100 microM GLU did enhance the stimulated release of GABA. The cholinergic agonist, carbachol, enhanced the stimulated release of NE, 5-HT and GLU 10, 60 and 40%, respectively. On the other hand, carbachol attenuated the release of DA and GABA approximately 20%. One interpretation of the data is that the amino acid transmitter pathways in slices of the cerebral cortex of the rat can be controlled by monoaminergic and cholinergic systems while the monoamine afferents appear to have a cholinergic regulation but not a major direct amino acid transmitter influence.     

An analysis of the axon populations in the nerves to the pelvic viscera in the rat

The present study uses selective surgical ablations combined with electron microscopic analyses to determine the number of axons in yarious catagories in rat hypogastric, pelvic, and pudendal nerves, these being the nerves to the pelvic viscera in this animal. Unmyelinated fibers predominate in all of these nerves. One of the most significant findings is that the pelvic nerve contains almost as many postganglionic sympathetic fibers as the hypogastric nerve. Previous investigators thought that the pelvic nerve supplied the parasympathetic inflow and the hypogastric nerve the sympathetic inflow to the pelvic viscera. The finding that there is a sizable sympathetic component in the pelvic nerve negates this idea, at least for the rat, and presumably calls for a reevaluation of the syndromes that arise from pelvic as opposed to hypogastric nerve section. Other findings of interest are (1) that there are unmyelinated efferent axons in the pudendal nerve, indicating that the pudendal is not a typical somatic nerve, (2) that the hypogastric nerve has a very small sensory component, and (3) that there are fibers surviving in the distal stumps of all these nerves, particularly the pelvic and hypogastric nerves.     

Differential effects of progesterone and genital stimulation on sequential inhibition of estrous behavior and progesterone receptor expression in the rat brain

The effect of genital stimulation, either by vaginocervical stimulation (VCS) using a calibrated vaginal probe combined with manual flank stimulation (FS), or by mounts performed by the male, on the hypothalamus and preoptic area concentration of the progesterone receptors A (PR-A) and B (PR-B) was assessed in ovariectomized (ovx) estrogen-primed rats. VCS/FS or stimulation provided by male mounts, even without intromission, significantly decreased PR-B concentration in the hypoythalamus. Down regulation of PR produced by genital stimulation was quantitatively similar to that elicited by progesterone (P) administration. Bilateral or unilateral transection of the pelvic or the pudendal nerves prevented down regulation elicited by VCS/FS. Repeated VCS/FS elicited lordosis behavior in most ovx estrogen primed rats, but the lordosis intensity was lower than that observed in response to P. P administered to ovx estrogen primed rats, induced sequential inhibition, i.e., failure to display estrous behavior in response to a second P injection (24h after the initial P injection). VCS/FS failed to elicit sequential inhibition, since rats responded with normal estrous behavior to the second injection of P. This suggests that down regulation by VCS, by contrast with P, failed to inhibit the subpopulation of PR involved in the facilitation of estrous behavior by P.     

Differential roles of hypogastric and pelvic nerves in the analgesic and motoric effects of vaginocervical stimulation in rats

Bilateral transection of the pelvic and/or hypogastric nerves, which convey afferent activity from the reproductive tract, was performed to ascertain the role of these nerves in the analgesic and motoric effects of vaginocervical mechanostimulation (VS) in rats. Two indices of analgesia were used: tail flick latency to radiant heat (TFL) and vocalization threshold to electrical shock of the tail (Voc-T). Nerve cuts were performed at least one week prior to behavioral testing. Bilateral transection of both the pelvic and hypogastric nerves eliminated the analgesic effects of VS on the TFL and Voc-T tests. Bilateral transection of only the pelvic nerves reduced the number of rats showing maximal VS-induced elevation in TFL, without altering the effect of VS on Voc-T. By contrast, bilateral transection of only the hypogastric nerves attenuated the Voc-T-elevating effect of VS, without reducing the effect of VS on elevating TFL. The effects of VS on producing immobility, hindlimb extension and blockage of hindlimb withdrawal to foot pinch were eliminated by combined bilateral pelvic and hypogastric neurectomy. However, bilateral transection of either nerve alone did not significantly alter the efficacy of VS in producing these effects. These findings indicate that the pelvic and hypogastric nerves contribute to the immobility- and extensor-inducing, and flexor-inhibiting effects of VS, and differentially mediate the analgesia-producing effects of VS.     

Effect of nucleus tractus solitarius lesions on cardiovascular responses elicited from the caudal ventrolateral medulla.

Neurons in the caudal ventrolateral medulla (CVLM) which inhibit sympathetic vasomotor tone may have reciprocal connections with the nucleus tractus solitarius (nTS). This study determined whether changes in arterial pressure elicited by chemical excitation or inhibition of neurons in the vasodepressor region of the rabbit CVLM depend on the integrity of the nTS. Unilateral injections of L-glutamate (10 pmol to 100 nmol), or bilateral injections of GABA (1 nmol to 125 nmol), were made into the CVLM, and dose-response effects on arterial pressure determined. The nTS was then bilaterally cauterized, or inhibited by local injections of muscimol, and the dose-response curves were repeated. Neither cauterization nor injection of muscimol significantly altered the slope of the log dose-response curves for L-glutamate, but nTS muscimol increased the fall in arterial pressure for each dose of L-glutamate (P less than 0.01). Cauterization of the nTS significantly (P less than 0.01) increased the slope of the curve relating dose of GABA to rise in arterial pressure observed, after injection of GABA into the CVLM. This increase in slope was similar to the increase observed when GABA is injected into the CVLM in baroreceptor-denervated rabbits. We conclude that neither the depressor nor the pressor response evoked by stimulation or inhibition of the CVLM is dependent on the integrity of the nTS. Inactivation of the nTS tends to increase the magnitude of the CVLM responses, possibly by removal of baroreceptor-mediated buffering of the responses.     

Brain-mediated responses to vaginocervical stimulation in spinal cord-transected rats: role of the vagus nerves

The present study was designed to ascertain whether the vagus nerves convey functional sensory activity from the reproductive tract in rats. Previously, vaginocervical mechanostimulation (VS) was shown to increase pupil diameter (PD) and the threshold of vocalization to tail shock (Voc-T). These responses were attenuated but not abolished by combined bilateral transection of the ‘genito-spinal’ nerves (i.e. pelvic, hypogastric and pudendal). Subsequent bilateral vagotomy further reduced or abolished the residual responses. In the present study, spinal cord transection above the known level of entry of the genito-spinal nerves was combined with bilateral vagotomy. In ovariectomized rats, after spinal cord transection at thoracic 7 (T7X), lumbar 5 (L5X) levels, or sham surgery (Sh), responses to VS were measured, the vagus nerves were then transected bilaterally, and responses to VS were again measured. VS significantly increased Voc-T and PD after sham procedure or spinal cord transection at either level. Subsequent bilateral vagotomy abolished the VS-induced increase in PD in the T7X group. Due to low survival rate, the effect of vagotomy on Voc-T could not be determined. Consequently, we performed a second experiment. In non-ovariectomized rats, VS significantly increased PD but reduced Voc-T in the T7X group compared to the Sh group, and subsequent bilateral vagotomy abolished both responses. These findings provide evidence that, in the rat, the vagus nerves provide a functional sensory pathway from the reproductive tract directly to the medulla oblongata of the brain, bypassing the spinal cord.     

Ontogenic modifications in the effect of the GABAergic system on the hypothalamic excitatory amino acids: its relationship with GABAergic control of gonadotrophin secretion during sexual maturation in female rats

Aminooxyacetic acid (AOAA), an inhibitor of gamma-aminobutyric transaminase, stimulates the in vitro GABA release by medial and anterior preoptic hypothalamic areas in prepubertal female rats (6, 15 and 30 days of age). This increase of GABA release at 15 days of age, was accompanied by a significant increase (P<0.01) in the hypothalamic release of glutamate (GLU) and aspartate (ASP), the excitatory amino acids involved in N-methyl-D-aspartate neurotransmission and a decrease in the release of these excitatory amino acids at 6 and 30 days of age (P<0.01). The increase in the hypothalamic release of GLU and ASP at 15 days of age was accompanied by a significant increase of the plasmatic LH and FSH concentration, while the hypothalamic decrease of excitatory amino acids release induced by AOAA also decreased LH and FSH plasmatic levels at 6 and 30 days of age. In summary, the present results show that in female rats there are differences in the effect of GABAergic system the hypothalamic release of GLU and ASP and on gonadotrophin secretion at different ages of prepubertal period, i.e. an inhibitory effect at 6 and 30 days of age and a stimulatory one at 15 days of age. It is proposed that the different effects of GABA on gonadotrophin secretion in prepubertal rats previously described are connected with ontogenic changes in the interrelationships between GABAergic and NMDA neurotransmission systems during sexual maturation of the hypothalamus in female rats. It is probable that these ontogenic modifications are connected with the maturation of interneuronal connection and/or new receptors activity.     

Muscle fiber type morphology and distribution in paraplegic patients with traumatic cord lesion

Summary Previous work showed that lysolecithin injected into rat sciatic nerve produced axonal degeneration of non-myelinated fibres. The possibility was raised that the swollen axons observed proximal to the injection site were the cause rather than the result of axonal degeneration (Mitchell and Caren 1982).The two main objectives of the present study were to examine the effects of different concentrations of lysolecithin on a nerve composed mainly of nonmyelinated fibres and by histochemical means, to study axoplasmic transport in the lysolecithin exposed nerves.The guinea-pig inferior mesenteric ganglion (IMG) and its associated hypogastric nerves, which were ligated, was placed in a three compartment chamber and maintained in vitro for 24 h at 37° C. This in vitro preparation was chosen because the concentration of lysolecithin added to the hypogastric nerves could be accurately measured and axonal transport in the lysolecithin-exposed nerves could be studied using two different methods. Horseradish peroxidase (HRP) applied to the ligated nerve compartment was used to study retrograde transport and the endogenous noradrenaline (NA) content of the sympathetic hypogastric nerves was used to study orthograde transport.The lysolecithin-induced changes were dose dependent and similar to those observed previously in vivo. When the damage was restricted to Schwann cells both HRP and NA were transported along the axons in the lysolecithin-exposed segment of nerve. Only when the lysolecithin produced ultrastructural evidence of axonal degeneration was there any observed accumulation of HRP or NA.These results and the value of this preparation for studying the effects of different toxins on nonmyelinated fibres are discussed.Supported in part by an MRC project grant No. G. 979/622/N     

Roles of neuroactive amino acids in ammonia neurotoxicity

Many neurologic disorders are related to congenital or acquired hyperammonemia (HA). Advanced symptoms of HA range from seizures in acute stages to stupor and coma in more chronic conditions, manifesting variable imbalance between the inhibitory and excitatory neurotransmission. Evidence obtained with the use of experimental HA models suggests that acute neurotoxic effects of ammonia are mediated by overactivation of ionotropic glutamate (GLU) receptors, mainly the N-methyl-D-aspartate (NMDA) receptors, and to a lesser degree the KA/AMPA receptors. NMDA receptor–mediated neurotoxicity may be potentiated by impaired control of their function by metabotropic GLU receptors, which are inactivated by ammonia. Prolonged overactivation of the NMDA receptors upon extended ammonia exposure causes their downregulation. The GLU receptor changes may be related to their excessive exposure to extrasynaptic GLU. Ammonia promotes GLU accumulation in the extrasynaptic space by enhancing its release from neurons, and/or by decreasing its reuptake to the nerve endings and astrocytes, where the effect results from inactivation (downregulation) of the astrocytic glutamate transporter GLT1. Excitotoxic effects of ammonia are augmented by increased synthesis of nitric oxide (NO), which is associated with NMDA receptor activation and/or increased synaptic transport of arginine (ARG). A shift toward neural inhibition is promoted by positive modulation of the γ-aminobutyric acid (GABA)ergic tone resulting from excessive accumulation in the brain of endogenous central benzodiazepine receptor agonists, and from upregulation of astrocytic peripheral benzodiazepine receptors leading to elevated levels of prognenelone-derived neurosteroids, which positively modulate the GABA(A) receptor complex. Inhibitory neurotransmission may also be favored by enhanced release from astrocytes of an inhibitory amino acid, taurine. J. Neurosci. Res. 51:133–138, 1998. © 1998 Wiley-Liss, Inc.     

The effects of decentralization on substance P-immunoreactivity in the anterior major pelvic ganglion of the male guinea pig.

The anterior major pelvic ganglion (AMPG) of the male guinea pig possesses a substance P (SP)-immunoreactive peri-neuronal plexus. Selective nerve transections involving the principal inputs of the AMPG, the hypogastric and pelvic nerves, indicate that the SP-immunoreactive peri-neuronal plexus is derived from multiple sources: an extrinsic source involving both the hypogastric and pelvic nerves, and another source (possibly the projections of small intensely fluorescent cells). SP-immunoreactivity (IR) is not normally present in the neuronal perikarya of the AMPG. Evidence is presented that suggests the absence of SP-IR is due to an active suppression of SP-synthesis. This seems to be achieved by a trans-synaptic mechanism involving the hypogastric nerve which, after transection, leads to the appearance of neuronal perikarya exhibiting SP-IR (less than 1% of the total neuronal population of the AMPG). Up to 65% of the neuronal perikarya of the AMPG have the ability to synthesize SP, as demonstrated by SP-IR after 24 h in vitro. A more potent factor in the down-regulation of SP synthesis seems to be exerted by the pelvic genito-urinary organs, especially the prostate.