Are GAD65 and GAD67 associated with specific pools of GABA in brain?

Brain contains at least two pools of gamma-aminobutyric acid (GABA), the transmitter pool and the so-called metabolic pool. To a large extent these pools may reflect the presence of GABA in different intracellular compartments, as immunocytochemical studies show that GABA is not localized mainly in terminals but is distributed throughout neurons. An interesting issue is the extent to which the two major forms of glutamate decarboxylase (GAD65 and GAD67) are specialized to synthesize GABA for these pools. Although GAD65 and GAD67 differ significantly in several characteristics, they also have substantial similarities and interactions, and the presence of individual forms of GAD in certain cell types is consistent with the idea that GAD65 and GAD67 can each synthesize GABA for both pools. Substantial progress has been made in understanding the regulatory properties of GAD, but the available data provide little indication of how differences between the forms might enable each to serve the demands for GABA synthesis in a specific pool.

Modulation of morphine antinociception by the brain-penetrating H2 antagonist zolantidine: detailed characterization in five nociceptive test systems.

Because histamine (HA) in the CNS may be a mediator of antinociception, a detailed investigation of the effects of the brain-penetrating H2 antagonist zolantidine (ZOL) was performed on five nociceptive tests in the presence and absence of morphine (MOR) in rats. ZOL inhibited MOR antinociception on the tail flick test, although a diurnal difference (inhibition in the dark cycle >> light cycle) was found. Similar results were found with the hot plate test, although details of the test procedure were significant. In contrast, ZOL induced opposing effects on MOR antinociception on two nonthermal tests (jump test and tail pinch test); ZOL alone induced moderate antinociception on the former test and mild antinociception on the latter test. Thus, ZOL exerts differential effects on baseline nociception and on MOR antinociception that vary depending on the nociceptive test employed, the light-dark cycle of the subjects, and the degree of stress associated with the nociceptive testing. These complex effects reveal the heterogeneous nature of opiate-induced modulation of nociception, and show that ZOL is a powerful tool for studying the relationships between opiates, HA, and nociceptive mechanisms.

Histamine-mediated neuronal death in a rat model of Wernicke's encephalopathy.

Three experiments were conducted to examine the role of histamine in neuronal degeneration in a rat model of Wernicke's encephalopathy induced by an acute bout of pyrithiamine-induced thiamine deficiency (PTD). In the first experiment, histamine levels in medial thalamus of freely moving PTD rats measured by microdialysis were increased (180% of controls) at a prelesion stage of thiamine deficiency (treatment day 12) and further elevated 48 hr later (380%) in the same animals when necrosis was evident. Histamine levels in dialysates of the hippocampus collected simultaneously from the same animals were unchanged at either stage of thiamine deficiency. Glutamate levels in microdialysates from the same animals were unchanged at the prelesion stage but were significantly elevated on the second collection day. In a second experiment, separate groups of PTD and pairfed control (CT) rats were infused continuously with either alpha-fluoromethylhistidine (FMH; 80 mg/day, i.p.), an irreversible inhibitor of histamine synthesis, or saline. FMH pretreatment produced a significant protection against PTD-induced neuronal loss within the midline-intralaminar and anteromedial thalamic nuclei, but had no effect on damage to ventrolateral nuclei, anteroventral nucleus, or the mammillary bodies. In a third study, histamine (80 micrograms, free base) or vehicle was directly infused into the same region of medial thalamus dialyzed in experiment 1. Histamine infusion into prelesion PTD but not CT animals resulted in severe neuronal loss and gliosis. Infusion of vehicle into the same regions of PTD and CT rats produced a mild gliosis restricted to the needle tract with no evidence of neuronal loss. These observations together with recent evidence of a histamine enhancement of glutamate receptor activation suggest that early histamine release may contribute significantly to glutamate-N-methyl-D-aspartate (NMDA)-mediated excitotoxic neuronal death in thiamine deficiency-induced Wernicke's encephalopathy.

Characterization of basal and morphine-induced histamine release in the rat periaqueductal gray.

Previous studies have shown that antinociceptive doses of systemic morphine increase extracellular histamine (HA) levels in the rat periaqueductal gray (PAG), although the cellular origin of basal and morphine-induced HA release in the PAG is unknown. Treatment with alpha-fluoromethylhistidine (FMH; 100 mg/kg, i.p.), the irreversible inhibitor of histidine decarboxylase, decreased basal HA release by a maximum of 80% and prevented morphine-induced HA release in the PAG. In addition, perfusion of this area with the sodium channel blocker tetrodotoxin (10(-6) M) decreased basal HA release by a maximum of 57% from baseline levels. When the perfusion medium was modified by substitution of magnesium for calcium, extracellular HA levels in the PAG decreased by a maximum of 72%, and morphine-induced HA release was prevented. Thioperamide (5 mg/kg, i.p.), an H3 antagonist, increased HA release in the PAG to a maximum of 249% within the first 30-60-min period. Taken together, these results suggest that basal and morphine-induced HA release in the rat PAG have a neuronal origin.

Nicotine tolerance in chromaffin cell cultures: acute and chronic exposure to smoking-related nicotine doses.

Nicotine tolerance and dependence are key aspects of tobacco addiction; however, the cellular mechanisms underlying these phenomena are poorly understood. Adrenal chromaffin cells release catecholamines upon exposure to nicotine and with repeated exposure this response exhibits nicotine tolerance. Using bovine adrenal chromaffin cells in culture, we have demonstrated acute and chronic nicotine tolerance at doses relevant to that in the blood and tissues of smokers (10(-7) M to 10(-6) M). Chromaffin cells are preexposed to low doses of nicotine for time periods ranging from 10 min to 7 days and then subsequently challenged with a maximally stimulating dose of nicotine (10(-5) M) for 10 min, all at 37 degrees C. Preexposure to nicotine results in a depression of 45Ca uptake and catecholamine release upon subsequent nicotine challenge. Acute tolerance or desensitization of nicotine-stimulated catecholamine release begins to occur in minutes after preexposure to 10(-6) M nicotine at 37 degrees C. The depression of catecholamine release upon preexposure to nicotine is both dose and temperature dependent and is not seen with potassium-evoked release. Chronic exposure to 10(-7) M nicotine for 3 days led to a depression of the secretory response to approximately 70% of control responses. There was a trend toward recovery of full response by days 5 and 7 of 10(-7) M nicotine preexposure. Nearly complete depression of the nicotine-evoked release occurs within the first day of exposure to 10(-6) M nicotine and persists for at least a week of nicotine exposure at 37 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

Histamine does not play an essential role in electrocortical activation during waking behavior.

Intraperitoneal injection of alpha-fluoromethylhistidine (alpha-FMH; 200 mg/kg), a specific inhibitor of histidine decarboxylase produced a severe depletion of neocortical and hippocampal histamine 3 h later as determined by a radioenzymatic assay. This treatment had no obvious effect on either low voltage fast activity (LVFA) in the neocortex or on rhythmical slow activity (RSA) in the hippocampus during an 8 h recording period during the rats' light cycle. Scopolamine-sensitive LVFA, scopolamine-resistant LVFA and scopolamine-resistant hippocampal RSA all appeared unaffected. This suggests that any contribution histamine makes to electrocortical activation is probably indirect, acting via other transmitter systems.

alpha-Fluoromethylhistidine-induced inhibition of brain histidine decarboxylase. Implications for the CO2-trapping enzymatic method.

The actions of S-alpha-fluoromethylhistidine (FMH), an irreversible inhibitor of the histamine biosynthetic enzyme histidine decarboxylase (HD), were studied on rat brain HD, as measured by a recently developed CO2-trapping enzymatic method. As expected, FMH induced a virtually complete inhibition of HD in the hypothalamus both in vivo and in vitro. In the frontal cortex, however, maximal doses of FMH did not maximally inhibit HD, suggesting the existence of an FMH-resistant form of HD. Careful studies of the conditions under which the assays were performed (homogenate dilution, preincubation times, incubation times, temperatures), as well as experiments with inhibitors of other decarboxylases, were unable to provide an explanation for this. When comparable studies of the effects of FMH in these brain regions were performed by alternative methods for measuring HD activity, no evidence for the existence of an FMH-resistant form of HD could be found. Thus, even though the CO2-trapping method appears to be accurate for measuring HD activity in rat hypothalamic homogenates, the present results show that this method may not be specific when studying brain regions other than the hypothalamus.

Simultaneous measurement of opiate-induced histamine release in the periaqueductal gray and opiate antinociception: an in vivo microdialysis study.

Histamine release and the subsequent activation of H2 receptors in the periaqueductal gray (PAG) are thought to be important components of morphine antinociception. In vivo microdialysis and antinociceptive testing were simultaneously applied in rats to characterize the effects of morphine on PAG histamine release and determine the relationship between histamine release and antinociception. In the absence of nociceptive (tail pinch) testing, morphine (12.8 mg/kg) induced a delayed, long-lasting release of histamine in the PAG. This effect of morphine was abolished by the opiate antagonist naltrexone (1 mg/kg) but was not mimicked by the mu-preferring agonist fentanyl (0.3 mg/kg), suggesting that activation of an opiate receptor other than, or in addition to, the mu receptor is necessary. In contrast to the findings with fentanyl in untested animals, fentanyl combined with nociceptive testing increased histamine release, even though testing alone had no such effect. Unexpectedly, tail pinch testing inhibited morphine-induced histamine release. These results show that the test procedure alters the action of opiates on histamine release, an effect likely to be the result of the stress of repeated tail pinch testing. Therefore, although histamine release may not be obligatory for all types of opiate antinociception, histamine in the PAG may function as a mediator of stress-induced potentiation of opiate antinociception. Even though the microdialysis technique has been acclaimed for its ability to assess neurochemical and behavioral characteristics simultaneously, the introduction of nociceptive testing clearly can alter the neurochemical systems under study.

Opiates, mast cells and histamine release.

Opiates have long been known to cause the release of histamine from mast cells, resulting in several undesirable effects, such as hypotension, urticaria, pruritus, and tachycardia. The mechanism of this opiate response has remained unclear, although it is known to be non-immunological in nature. A survey of the histamine-releasing properties of a variety of opiates shows that the pharmacology of opiate-induced histamine release from mast cells is distinct from that of known opiate receptors. Although functional opiate receptors may exist on mast cells and may be capable of modulating IgE-mediated histamine release, there is no evidence that these receptors account for opiate-induced histamine release. Since other basic compounds have been suggested to release histamine from mast cells by directly activating G-proteins, it seems possible that morphine and endogenous opiates may also share this mechanism.

Morphine-induced increases of extracellular histamine levels in the periaqueductal grey in vivo: a microdialysis study.

The effect of morphine on extracellular histamine levels in two regions of the rat midbrain was studied in vivo by microdialysis. Morphine (5.6 and 12.8 mg/kg, s.c.) significantly and dose-dependently increased extracellular histamine levels in the periaqueductal grey, while no significant effect was observed in the reticular formation. In addition, no significant effect of sequential saline injections was observed on extracellular histamine levels in the periaqueductal grey. Since morphine has no effect on histamine catabolism, these results suggest that morphine increases histamine release in the rat PAG, a site where morphine and histamine are known to have analgesic action. Taken with earlier studies showing the ability of H2 antagonists to block morphine analgesia, these results support the hypothesis that histamine and H2 receptors are important in mediating morphine analgesia in the rat periaqueductal grey. The cellular origin of the extracellular histamine, and the mechanism of this morphine effect remain to be determined.