Possible mechanisms of the effect of physostigmine on the facilitation of acetylcholine release in the guinea pig myenteric plexus

The automodulation of acetylcholine (ACh) release in the guinea pig myenteric plexus-longitudinal muscle preparation was investigated by studying the electric stimulation-evoked release of radiolabeled ACh. When the release associated with neuronal activity was challenged by the muscarinic antagonist atropine, the release was not significantly enhanced. When the acetylcholinesterase (AChE) blocker physostigmine was present, the well-established muscarinic receptor-mediated autoinhibition was operative i.e., the release was significantly reduced. However, when both drugs were added together, the release was much higher than under control conditions. Therefore, it seems likely that there is also a facilitatory system. We made an effort to clear up the mechanism of this facilitation by blocking possible nicotinic presynaptic receptors, by excluding the α₂-adrenoceptor-mediated masking effect of noradrenergic heteromodulation, by preventing a possible ATP-mediated mechanism, and by attempting to prevent the direct effect of physostigmine. None of these manipulations resulted in a decrease of the surplus release. It is concluded, that when the negative feedback modulation of ACh is inhibited and AChE activity is reduced, high levels of ACh facilitates additional release of ACh from the nerve terminals, possibly through a not yet verified class of receptors.     

Developmental change of cholinergic activity in the forebrain of the zebra finch during song learning.

Song learning in the male zebra finch closely correlates with the development of song-control nuclei, the high vocal center (HVc), the robust nucleus of the archistriatum (RA) and the lateral magnocellular nucleus of the anterior neostriatum (IMAN). We previously found that acetylcholine (ACh) content increases transiently in the RA during the sensitive period of song learning. In the present experiment, development of cholinergic neurons by immunohistochemical methods using monoclonal choline acetyltransferase (ChAT) antibodies, and enzymic activity of ChAT and acetylcholinesterase (AChE) were examined in the RA during the sensitive period of song learning. The developmental change of ChAT activity in the RA was closely parallel to that of the ACh concentration. On the other hand, the density of ChAT-immunoreactive fibers changed little in the RA. This indicates that the transient increase of ACh content during the sensitive period is due to ChAT activity, but not to the increasing density of the cholinergic fibers. The ACh in the RA might be related to plastic changes in the synapse of RA neurons.     

Pharmacology of the brachium conjunctivum: red nucleus synaptic system in the baboon.

Acetylcholine, biogenic amines, and certain amino acids were applied by microiontophoresis to parvicellular and magnocellular red nucleus (RN) neurons of baboon while recording brachium conjunctivum (BC)-evoked and amino acid-evoked unit discharge from these neurons. Glycine, gamma-aminobutyric acid, and beta-alanine were potent depressants of BC-RN synaptic transmission, amino acid-evoked firing, and spontaneous activity of all RN neurons studied. Glycine was clearly more potent than the other 2 depressant amino acids. L-Glutamic and DL-homocysteic acid were strong excitants of all RN neurons tested. Dopamine, noradrenaline, and 5-hydroxytryptamine depressed the excitability of both parvicellular and magnocellular RN neurons; no excitatory effects were observed with these biogenic amines on RN neurons. Acetylcholine increased the rate of firing of spontaneously discharging parvicellular RN neurons and facilitated the amino acid-induced firing of these same neurons. Acetylcholine did not facilitate BC-RN synaptic transmission nor could this transmission be blocked by cholinergic antagonists. Unlike parvicellular RN neurons, the responsiveness of magnocellular neurons was either unaltered by acetylcholine or slightly decreased. These experiments demonstrate a difference in the pharmacologic responsiveness of parvicellular and magnocellular RN neurons to acetylcholine but do not provide evidence for a cholinergic input to RN via the brachium conjunctivum.     

No triazolam-induced expression of Fos protein in raphe nuclei of the male Syrian hamster

While the visual projections to the suprachiasmatic nuclei (SCN) play a role in mediating the effects of light on circadian rhythms, the functional significance of the serotonergic projection from the raphe nuclei (RN) to the SCN is uncertain. Because previous results indicated that RN would appear to be a likely site for triazolam (Tz)-induced phase shifts, we used the expression of Fos-protein as a marker of Tz-induced neuronal activation. Immunocytochemistry was used to visualize the presence of Fos-like protein. Tz-induced Fos-labeled nuclei were found in superior colliculi. Edinger-Westphal nuclei (EW) and dorsal tegmental nuclei (DTg), but not in the RN. The SCN showed only occassionally labeled nuclei in all experimental groups, whereas there was no Tz-induced Fos-immunoreactivity in the intergeniculate leaflet (IGL). The present data not necessarily exclude the implication of the RN in the phase shifting effect of TZ. The phase shift could still be accomplished using a different set of immediate early genes (IEG), or without an IEG response. Alternatively, as will be discussed, other pathways could mediate the phase shifting effect of Tz.     

Postnatal development of acetylcholinesterase in, and cholinergic projections to, the cat superior colliculus.

The postnatal development of cholinergic afferents to the superior colliculus in neonatal cats was studied by using acetylcholinesterase (AChE) histochemistry, choline acetyltransferase (ChAT) immunohistochemistry, and retrograde transport of horseradish peroxidase (HRP). In the adult cat, the pattern of AChE staining was laminar specific. AChE was distributed continuously in the stratum griseum superficiale (SGS) but was organized as patches in the stratum griseum intermediate (SGI). Diffuse AChE staining also was present in the stratum griseum profundum (SGP) and the dorsolateral periaqueductal gray (PAG). At birth, however, AChE staining was barely detectable in the SGS and, aside from a few isolated labeled neurons, was absent from the SGI, SGP, and PAG. By 7 days postnatal (dpn), staining in the SGS was more apparent but did not change appreciably in the deeper laminae. A substantial increase in AChE staining occurred in the SGS at 14 dpn (several days after eye opening), at which time patches in the SGI first became apparent. By 28 dpn, the complete laminar-specific adult AChE staining pattern was present, though the staining intensity did not reach the adult level until 56 dpn. A protracted maturation of both AChE staining and ChAT immunoreactivity also was observed in the sources of cholinergic afferents to the superior colliculus, which include the parabigeminal nucleus, and the pedunculopontine (PPN) and lateral dorsal tegmental (LDTN) nuclei. AChE and ChAT-immunoreactive staining in each nucleus was weak at birth but increased during the ensuing 2 weeks. At 21 dpn, however, ChAT immunoreactivity virtually disappeared in the parabigeminal nucleus and significantly decreased in PPN and LDTN. The ChAT immunoreactivity in these nuclei then gradually increased reaching maximum levels by 28 dpn. At 35 dpn, AChE staining showed a significant, though temporary (4 weeks), decrease in the parabigeminal nucleus, but not in the PPN and LDTN, that subsequently increased to the adult level of staining at 70 dpn. The absence of AChE in the SGI in neonatal animals was correlated, at least in part, with a paucity of neurons in the brainstem cholinergic cell groups labeled by retrograde transport of HRP from the superior colliculus. Injections of HRP into the superior colliculus retrogradely labeled many neurons in the parabigeminal nucleus, but few, if any, neurons in the PPN or LDTN at 1 dpn. Retrogradely labeled neurons also were observed in the substantia nigra pars reticulata, albeit fewer in neonates than in adults.(ABSTRACT TRUNCATED AT 400 WORDS)     

A subchronic application period of glucocorticoids leads to rat cognitive dysfunction whereas physostigmine induces a mild neuroprotection

The cholinergic neurotransmitter system and prolonged glucocorticoid-induced stress can affect cognitive functions in opposite ways. While pharmacological enhancement of cholinergic neurotransmission is known to induce neuroprotective effects, chronic glucocorticoids impair cognitive functions. Up to now, there is no consensus as to whether a subchronic stress period of several days would affect cognitive function. The goal of this study was to investigate whether or not repeated applications of physostigmine over 3 days lead to protective effects on rat spatial cognitive abilities in contrast to the deteriorating effect on rat cognitive function after corticosterone treatment. Furthermore, we wanted to investigate in what extent this cognition-modulating effect is associated with rat cerebral acetylcholinergic system. Male adult rats (n = 40) were randomly divided into four groups with n = 10 per group: (I) placebo-, (II) corticosterone- (15 mg/day), (III) physostigmine- (0.014 mg/day), and (IV) physostigmine + corticosterone-treated rats. Body mass and plasma corticosterone concentrations were measured. Psychometric investigations were conducted using a Morris water maze before and after a subchronic treatment. In cerebral tissue, ACh and acetylcholinesterase (AChE) content and ACh receptor density were determined. Tissue corticosterone concentration was measured in cerebral cortex, hippocampus, and adrenal glands. In corticosterone-treated rats, reduced spatial cognitive abilities were associated with a significant increase in plasma (+25%) and cerebral corticosterone levels (+350%) parallelled by a significant reduction in adrenal gland concentrations (−84%) as compared to placebo. Repeated physostigmine injections improved rats’ spatial memory and increased cerebral ACh and AChE content (p < 0.05). When physostigmine was administered at the same time as corticosterone (group IV), it was not able to reverse the corticosterone effect. A significant correlation was detected between cerebral AChE and corticosterone concentrations as well as between AChE and psychometric parameters. We conclude that subchronic exogenous corticosterone administration induces memory dysfunction whereas physostigmine exerts cognitive-enhancing effects if given for 3 days. An apparently existing interaction between glucocorticoid excess and ACh metabolism is discussed.     

Serotonin selectively attenuates glutamate-evoked activation of noradrenergic locus coeruleus neurons.

The effect of 5-HT on activity of noradrenergic locus coeruleus (LC) neurons was studied using microiontophoretic and micropressure drug application in anesthetized rats. 5-HT had no consistent effect on LC spontaneous discharge, eliciting a modest decrease overall. However, 5-HT reliably attenuated responses of LC neurons to excitatory amino acids (EAAs), one of the major classes of transmitters in afferents to these neurons. This effect was specific for EAA responses because it occurred for glutamate and kainate but not for ACh. In contrast, iontophoretic norepinephrine (NE) selectively attenuated spontaneous activity but not responses evoked by either glutamate or ACh. The responsiveness of LC neurons to EAAs as quantified by a response-contrast measure (evoked excitation/basal activity) was markedly reduced by 5-HT, but was increased by NE. For ACh, such responsiveness of LC cells was not changed by 5-HT, but was increased by NE. The effects of 5-HT were prevented and reversed by iontophoretically applied antagonists of 5-HT receptors, methysergide and methiothepin. Thus, 5-HT appears to selectively interact with EAA responses of LC neurons, acting as a filter to attenuate LC activity linked to its major EAA inputs while allowing other channels afferent to the LC (e.g., those utilizing ACh) to be expressed.     

Acetylcholine and acetyl-CoA metabolism in differentiating SN56 septal cell line.

The rate of acetylcholine (ACh) synthesis was found to depend on the activity of choline acetyltransferase (ChAT) and on the concentrations of the two substrates of this enzyme, choline and acetyl-CoA. In SN56 cells treated for 3 days with 1 mM dbcAMP activities of ChAT and acetylcholinesterase (AChE) were elevated. It was accompanied by an increased activity of ATP-citrate lyase (ACL)-an enzyme responsible for provision of part of acetyl-CoA for ACh synthesis in cholinergic neurons. In contrast lactate dehydrogenase (LDH) and pyruvate dehydrogenase (PDH) activities were reduced by dbcAMP. Treatment with 0.001 mM all-trans retinoic acid (RA) elevated ChAT and LDH activities but reduced the activities of AChE and ACL. The combined treatment with db-cAMP and tRA increased ChAT activity in supra-additive fashion. The effects of these two compounds on the other enzymes were not additive. Neither compound altered the activities of carnitine acetyl-transferase, acetyl-CoA synthase, or acetyl-CoA hydrolase. On the other hand, they decreased acetyl-CoA content and rate of ACh release. Overall, the results indicate that tRA upregulates only ChAT expression, whereas dbcAMP upregulates several features of cholinergic neurons including ChAT, AChE, and ACL. Low levels of acetyl-CoA in differentiated cells may result in a low rate of ACh release and resynthesis during their depolarization.     

Denervation supersensitivity in the cholinergic septo-hippocampal pathway: a microiontophoretic study.

The purpose of this study was to determine whether denervation supersensitivity could be produced in an identified cholinergic pathway in the CNS of the rat. The mechanism for the development of this phenomenon was also explored. Cholinergic denervation of the hippocampus was accomplished by lesions of the medial septum. The response of hippocampal pyramidal cells to microiontophoretic application of acetylcholine (ACh) and carbachol in lesioned and unlesioned animals was determined by extracellular recording. There was a marked increase (6 X) in sensitivity to ACh 2-43 days following lesions. However, there was no increase in sensitivity to carbachol or glutamate. Other workers have shown that septal lesions cause a large decrease in hippocampal acetylcholinesterase (AChE) which is located mainly presynaptically. The absence of increase in sensitivity to carbachol, a cholinomimetic resistant to hydrolysis by AChE, suggests that the postlesion increase in sensitivity to ACh results from a decrease in its inactivation by AChE. A time course for the development of ACh supersensitivity was found to be similar to the time course of AChE loss previosly reported. Experiments using physostigmine, an AChE inhibitor, demonstrated that inhibition of AChE can potentiate the effects of ACh in unlesioned preparations, but not in lesioned preparations. We conclude: (1) denervation supersensitivity to ACh occurs in the septo-hippocampal pathway; and (2) the supersensitivity is probably due to decreased inactivation of ACh by AChE. The results suggest that presynaptic AChE plays a significant role in modulating the neurotransmitter function of ACh in the septo-hippocampal pathway.     

Effects of endogenous acetylcholine on spontaneous activity in rat dorsal cochlear nucleus slices

We have examined the contribution of endogenous acetylcholine (ACh) release to the spontaneous firing of both regular (probably fusiform cells) and bursting neurons (probably cartwheel cells) in the dorsal cochlear nucleus (DCN) in rat brainstem slices. The muscarinic antagonists atropine, scopolamine, and tropicamide (1–2 μM) caused substantial decreases of firing rates in a majority of the neurons. Reversible acetylcholinesterase (AChE) inhibitors typically caused large transient increases in firing that decayed more slowly than responses to carbachol. The irreversible AChE inhibitor diisopropyl fluorophosphate (DFP) usually caused a sustained increase, with an initial peak followed by a gradual change to a final level higher than before DFP. Tropicamide caused large decreases in firing after DFP, confirming sustained ACh release. Both neostigmine and DFP applied after AChE inhibition by DFP sometimes elicited a transient response. We conclude that the level of sustained response to DFP is determined by the rate of endogenous ACh release, and that DFP and reversible AChE inhibitors exert an initial transient agonist effect that overlaps the initial effect of acetylcholinesterase inhibition. The slice experiments provide a model for cholinergic mechanisms in vivo, confirm that the release of endogenous ACh increases the firing rates of regular and bursting neurons in superficial DCN, and support the hypothesis that spontaneous firing of DCN neurons is sustained in part by cholinergic inputs.     

Red nucleus neurons actively contribute to the acquisition of classically conditioned eyelid responses in rabbits

The red nucleus (RN) is a midbrain premotor center that has been suggested as being involved in the acquisition and/or performance of classically conditioned nictitating membrane/eyelid responses. We recorded in rabbits the activity of RN and pararubral neurons during classical eyeblink conditioning using a delay paradigm. Neurons were identified by their antidromic activation from contralateral facial and accessory abducens nuclei and by their synaptic activation from the ipsilateral motor cortex (MC) and the contralateral cerebellar interpositus (IP) nucleus. For conditioning, we used a tone as a conditioned stimulus (CS) followed 250 ms later by a 100 ms air puff as an unconditioned stimulus (US) coterminating with it. Conditioned responses (CRs) were determined from the evoked changes in the electromyographic activity of the orbicularis oculi (OO) muscle. Recorded neurons were classified by their antidromic activation and by their changes in firing rate during the CS-US interval. Identified neurons increased their firing rates in relation to the successive conditioning sessions, but their discharge rates were related more to the EMG activity of the OO muscle than to the learning curves. Reversible inactivation of the IP nucleus with lidocaine during conditioning evoked a complete disappearance of both conditioned and unconditioned eyelid responses, and a progressive decrease in CR-related activity of RN neurons. In contrast, MC inactivation evoked a decrease in the acquisition process and an initial disfacilitation of neuronal firing (which was later recovered), together with the late appearance of CRs. Thus, RN neurons presented learning-dependent changes in activity following MC inactivation.     

The experimental Alzheimer drug phenserine: preclinical pharmacokinetics and pharmacodynamics

Phenserine, a phenylcarbamate of physostigmine, is a new potent and highly selective acetylcholinesterase (AChE) inhibitor, with a >50-fold activity versus butyrylcholinesterase (BChE), in clinical trials for the treatment of Alzheimer's disease (AD). Compared to physostigmine and tacrine, it is less toxic and robustly enhances cognition in animal models. To determine the time-dependent effects of phenserine on cholinergic function, AChE activity, brain and plasma drug levels and brain extracellular acetylcholine (ACh) concentrations were measured in rats before and after phenserine administration. Additionally, its maximum tolerated dose, compared to physostigmine and tacrine, was determined. Following i.v. dosing, brain drug levels were 10-fold higher than those achieved in plasma, peaked within 5 min and rapidly declined with half-lives of 8.5 and 12.6 min, respectively. In contrast, a high (>70%) and long-lasting inhibition of AChE was achieved (half-life >8.25 h). A comparison between the time-dependent plasma AChE inhibition achieved after similar oral and i.v. doses provided an estimate of oral bioavailability of 100%. Striatal, in vivo microdialysis in conscious, freely-moving phenserine-treated rats demonstrated >3-fold rise in brain ACh levels. Phenserine thus is rapidly absorbed and cleared from the body, but produces a long-lasting stimulation of brain cholinergic function at well tolerated doses and hence has superior properties as a drug candidate for AD. It selectively inhibits AChE, minimizing potential BChE side effects. Its long duration of action, coupled with its short pharmacokinetic half-life, reduces dosing frequency, decreases body drug exposure and minimizes the dependence of drug action on the individual variations of drug metabolism commonly found in the elderly.     

Regulation of acetylcholine release by muscarinic receptors at the mouse neuromuscular junction depends on the activity of acetylcholinesterase

Muscarinic acetylcholine receptors (mAChRs) play an important role in regulating the release of acetylcholine (ACh) in various tissues. We used subtype-specific antibodies and a fluorescent-labelled muscarinic toxin to demonstrate that mammalian neuromuscular junction expresses mAChR subtypes M1 to M4, and that localization of all subtypes is highly restricted to the innervated part of the muscle. To elucidate the roles of the mAChR subtypes regulating ACh release, we measured the mean quantal content of endplate potentials in isolated mouse phrenic--hemidiaphragm preparations in which release was reduced by a low Ca2+/high Mg2+ medium. Muscarine decreased evoked ACh release in normal junctions but, depending on the concentration, reduced or increased transmitter release in collagen Q-deficient junctions completely lacking acetylcholinesterase (AChE). Both effects were also seen in normal junctions when AChE was inhibited by various doses of fasciculin-2. Block of mAChRs by atropine had no effect on evoked release at normal junctions, but decreased release at junctions lacking AChE. The muscarine-elicited depression of ACh release in normal junctions was completely abolished by pertussis toxin or methoctramine pretreatment, but was not affected by muscarinic toxin MT-3, thus indicating the involvement of the M2 mAChR. The muscarine-induced increase of ACh release in AChE-deficient junctions was not affected by pertussis toxin, but was completely blocked by MT-7, a specific M1 mAChR antagonist. Our results show that the M1 and M2 mAChRs have opposite presynaptic functions in modulating quantal ACh release, and that regulation of release by the two receptor subtypes depends on the functional state of AChE at the neuromuscular junction.     

Brachium conjunctivum-red nucleus synaptic system in the baboon

Electrophysiological properties of the brachium conjunctivum-red nucleus (BR-RN) synaptic system were studied in barbiturate-anesthetized baboon (Papio). Topographic recordings from the mesencephalon demonstrated that most of the BC-evoked activity was restricted to an area histologically identified as red nucleus; however, some brachium-evoked activity was recorded from the surrounding mesencephalic reticular formation. Short latency BC-evoked activity was also recorded from the pons in the region of nucleus reticularis tegmenti pontis. The majority of the BC fibers were found to conduct at a rat of 44 m/sec; a second group of BC fibers with a slower conduction velocity of 23 m/sec was also observed. Brachium-evoked responses recorded from magnocellular and parvicellular RN neurons were short latency responses consistent with monosynaptic activation of these RN neurons by the BC fibers. The BC-RN synaptic system was found to be a very secure synaptic system and could transmit activity at high rates of stimulation with little or no failure. The responsiveness of the BC fibers was found to be similar to that of optic nerve fibers and pyramidal tract fibers, both of which have been characterized as being similar to peripheral A fibers. The responsiveness of the BC-RN synaptic system began to decrease 5 msec after a single or repetitive transmission and was reduced to about 50% of normal responsiveness at 34 msec. This period of reduced postsynaptic responsiveness was associated with a reduction in presynaptic input to RN and suggest that a disfacilitation at the level of the deep cerebellar nuclei may be in part responsible for the subnormal responsiveness of the BC-RN synaptic system.     

Role of the parvicellular reticular formation in facilitating the jaw-opening reflex in rats by stimulation of the red nucleus

In a previous study, we have shown that electrical and chemical stimulation of the red nucleus (RN) facilitates the jaw-opening reflex (JOR). The RN sends projection fibers bilaterally, with contralateral dominance, to the part of the parvicellular reticular formation (RFp) containing premotor neurons projecting to the trigeminal motor nucleus. This implies that RN-induced facilitation of the JOR might be mediated via last-order neurons in the RFp. Here, we address this issue by investigating whether microinjection of lidocaine or l-glutamate into the RFp affects RN-induced modulation of the JOR. Experiments were performed on rats anesthetized with urethane-chloralose. The JOR was evoked by electrical stimulation of the inferior alveolar (IA) nerve and was recorded as an electromyographic response from the anterior belly of the digastric muscle. Conditioning stimulation was delivered unilaterally to the RN 12 ms before the IA test stimulation. We found that local injections of 2% lidocaine (0.5 microl) into the RFp, contralateral to the RN, significantly (P < 0.05) reduce RN-induced facilitation of the JOR, whereas corresponding injections of 0.1 mM l-glutamate (0.5 microl) significantly (P < 0.05) increase it. These results suggest that the facilitatory effect of RN stimulation on the JOR is mediated partly by a relay in the RFp.     

Neuroprotective and neurodestructive functions of nitric oxide after spinal cord hemisection

Nitric oxide (NO) may subserve different functions in different central neurons subjected to axotomy. The difference may depend on whether the neurons basally express neuronal nitric oxide synthase (nNOS), a biosynthetic enzyme of NO. This is supported by our previous finding that suggests the differential role of NO in neurons of nucleus dorsalis (ND) and red nucleus (RN) which have different basal expression of nNOS. This study aimed to establish firmly the functions of NO, as revealed by nNOS immunoreactivity and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry, by the administration of endogenous NO donor, l-arginine (l-arg), and NOS inhibitor, l-N(G)-nitroarginine methyl ester (l-NAME). To relate the role of NO to glutamate receptors (GluR), the distributions of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and N-methyl-d-aspartate receptor (NMDAR) in the two nuclei were revealed by immunohistochemical techniques. nNOS immunoreactivity was void in ND neurons, but expressed weakly in the RN normally. It was induced in ipsilateral ND neurons and upregulated on both sides of RN after spinal cord hemisection. Neuronal loss in the ipsilateral ND was augmented by l-arg, but reduced by l-NAME. In the contralateral RN, l-arg attenuated neuronal loss. NMDAR1 was present in most neurons in ND. After axotomy, some NMDAR1 immunoreactive neurons of the ipsilateral ND were induced to express NOS, whereas RN neurons showed strong staining for NMDAR1 and all the AMPA subunits. Most of the NOS-positive neurons in the RN were coexistent with GluR2 in normal rats and those subjected to axotomy. The present data demonstrated that NO exerted neurodestructive function in the non-NOS-containing ND neurons characterized by NMDAR as the predominant glutamate receptor. NO might be beneficial to the NOS-containing RN neurons. This could be attributed to the presence of GluR2. Possible diverse synthesizing pathways of NO in two different central nuclei were suggested from the observation that NOS was colocalized with NADPH-d in ND neurons, but not in RN neurons.     

红核在大脑皮质下行调控脊髓伤害感受性传递中的作用

以电刺激外周感受野诱发的大鼠脊髓背角WDR和NS神经元的晚串放电（C-反应）为指标，以串脉中刺激对侧大脑脚（CP）作为条件刺激，在C-反应受到明显抑制的神经元。分别观察了电解损毁红核（RN）和RN内注射兴奋性氨基酸的受体拮抗剂对刺激CP的下行抑制作用的影响。结果发现：损毁同侧RN后，刺激CP对C反应的抑制作用明显减弱，而损毁同侧RN背侧结构，对侧RN及假损毁RN均无此效应；RN内微量注射兴奋性氨基酸受体拮抗剂AP5和DNQX均可减弱刺激CP对C-反应的抑制。提示RN至少部分参与大脑皮质对脊髓伤害感受性传递的下行抑制作用。且以同侧RN为主；在与痛觉调制有关的皮质-RN通路中既有NMDA受体又有非NMDA受体的参与。     

Modulatory influences of red nucleus stimulation on the somatosensory responses of cat trigeminal subnucleus oralis neurons

Little is known of the effect of red nucleus (RN) stimulation on somatosensory neurons despite its known anatomic projections to somatosensory relay nuclei. The effect of RN stimulation on the somatosensory responses of trigeminal subnucleus oralis (Vo) neurons was investigated in chloralose- or barbiturate-anesthetized cats. Arrays of bipolar stimulating electrodes were inserted into the contralateral and ipsilateral RN and the contralateral thalamus. Extracellular single-unit recordings were obtained in Vo with tungsten microelectrodes. Neurons in Vo were excited to just suprathreshold by electrical stimulation within their receptive fields. Red nucleus influences were studied by applying 100-ms, 500-Hz conditioning trains to the contralateral or ipsilateral RN 130 ms prior to the peripheral test stimulus. The effect of RN stimulation was also tested on mechanically evoked responses of Vo cells. The somatosensory responses of most cells (70/73) were inhibited after RN stimulation. Some of these cells (15/70) could be antidromically activated from the contralateral thalamus. Stimulation of the RN resulted in excitation followed by inhibition in nine Vo cells. The results suggest that the RN may modulate transmission of somatosensory information through Vo.     

Effects of chronic descending tractotomy on the response patterns of neurons in the trigeminal nuclei principalis and oralis.

Unilateral, descending, trigeminal tractotomy was carried out on adult cats using aseptic technique. Unit activity, evoked by bipolar noxious stimulation of the tooth pulp and by innocuous mechanical stimulation of other receptive fields, was recorded from neurons in the trigeminal principalis and oralis nuclei, 7 to 13 weeks after tractotomy. Seventy neurons from the lesion side of the brain stem and 59 from the control side were studied. All units responded to pulpal stimuli and the great majority to mechanical stimuli. Three general response configurations were produced by each of the two forms of stimulation: brief bursts of 1 to 5 spikes, trains of 6 to 20 spikes, and multiple component discharges consisting of sequences of brief bursts. No statistically significant differences between the two populations were found for peripheral field size or location, for discharge characteristics such as latency, configuration, or mean density, or for responsiveness to the two types of stimuli. However, a statistically greater proportion of control units fired dense impulse trains (more than eight spikes) to intense pulpal stimuli. The results provide evidence that the response patterns of most somatosensory neurons in nuclei principalis and oralis are unaffected by descending trigeminal tractotomy and that stimulus modality is not coded by the response characteristics of polymodal units in these nuclei. Although some modulatory influence is exerted on polymodal neurons in the rostral nuclei by the trigeminal nucleus caudalis, it appears doubtful that this is a significant factor in determining whether a peripheral stimulus is perceived as noxious or innocuous.     

Acetylcholine from the mesopontine tegmental nuclei differentially affects methamphetamine induced locomotor activity and neurotransmitter levels in the mesolimbic pathway

Methamphetamine (MA) increases dopamine (DA) levels within the mesolimbic pathway and acetylcholine (ACh), a neurotransmitter known to increase DA cell firing and release and mediate reinforcement, within the ventral tegmental area (VTA). The laterodorsal tegmental (LDT) and pedunculopontine tegmental (PPT) nuclei provide cholinergic input to the VTA; however, the contribution of LDT- and PPT-derived ACh to MA-induced DA and ACh levels and locomotor activation remains unknown. The first experiment examined the role of LDT-derived ACh in MA locomotor activation by reversibly inhibiting these neurons with bilateral intra-LDT microinjections of the M2 receptor agonist oxotremorine (OXO). Male C57BL/6 J mice were given a bilateral 0.1 μl OXO (0, 1, or 10 nM/side) microinjection immediately prior to IP saline or MA (2 mg/kg). The highest OXO concentration significantly inhibited both saline- and MA-primed locomotor activity. In a second set of experiments we characterized the individual contributions of ACh originating in the LDT or pedunculopontine tegmental nucleus (PPT) to MA-induced levels of ACh and DA by administering intra-LDT or PPT OXO and performing in vivo microdialysis in the VTA and NAc. Intra-LDT OXO dose-dependently attenuated the MA-induced increase in ACh within the VTA but had no effect on DA in NAc. Intra-PPT OXO had no effect on ACh or DA levels within the VTA or NAc, respectively. We conclude that LDT, but not PPT, ACh is important in locomotor behavior and the cholinergic, but not dopaminergic, response to systemic MA.