Extracellular Microelectrode Research Articles

Blockade of Nogo Receptor Ligands Promotes Functional Regeneration of Sensory Axons after Dorsal Root Crush

A major impediment for regeneration of axons within the CNS is the presence of multiple inhibitory factors associated with myelin. Three of these factors bind to the Nogo receptor, NgR, which is expressed on axons. Administration of exogenous blockers of NgR or NgR ligands promotes the regeneration of descending axonal projections after spinal cord hemisection. A more detailed analysis of CNS regeneration can be made by examining the growth of specific classes of sensory axons into the spinal cord after dorsal root crush injury. In this study, we assessed whether administration of a soluble peptide fragment of the NgR (sNgR) that binds to and blocks all three NgR ligands can promote regeneration after brachial dorsal root crush in adult rats. Intraventricular infusion of sNgR for 1 month results in extensive regrowth of myelinated sensory axons into the white and gray matter of the dorsal spinal cord, but unmyelinated sensory afferents do not regenerate. In concert with the anatomical growth of sensory axons into the cord, there is a gradual restoration of synaptic function in the denervated region, as revealed by extracellular microelectrode recordings from the spinal gray matter in response to stimulation of peripheral nerves. These positive synaptic responses are correlated with substantial improvements in use of the forelimb, as assessed by paw preference, paw withdrawal to tactile stimuli and the ability to grasp. These results suggest that sNgR may be a potential therapy for restoring sensory function after injuries to sensory roots.

Mechanisms of the facilitation of neurotransmitter secretion in strontium solutions

Experiments on frog neuromuscular synapses using extracellular microelectrode recording of endplate currents (EPC) and nerve ending (NE) responses were performed to study the mechanisms of facilitation of quantum secretion of acetylcholine on replacement of extracellular Ca ions with Sr ions. Solutions with a Ca ion concentration of 0.5 mM (calcium solutions) or a Sr ion concentration of 1 mM (strontium solutions) were used; the basal levels of neurotransmitter secretion (in conditions of low-frequency stimulation) were essentially identical. In calcium solutions, the drop in EPC facilitation on paired-pulse stimulation as the interimpulse interval was increased from 5 to 500 msec was described by the sum of three exponential components - the early, the first, and the second. In strontium solutions, facilitation was decreased as compared with the level in calcium solutions predominantly because of decreases in the early and first components. At the same time, EPC facilitation in conditions of rhythmic stimulation (10 or 50 impulses/sec) in strontium solution was significantly increased as compared with the level in calcium solutions. In strontium solutions in conditions of high-frequency stimulation at 50 impulses/sec, there was also a marked decrease in the amplitude of the third phase of the NE response, reflecting NE potassium currents. These data lead to the conclusion that the facilitation sites underlying the first and early components had lower affinities for Sr ions than for Ca ions. Increases in facilitation in strontium solutions in conditions of high-frequency rhythmic activity resulted from two mechanisms: more marked widening of the NE action potential and an increase in the divalent cation influx current due to weak activation of the Ca2+-dependent potassium current in the presence of Sr ions, as well as the slow dynamics of the removal of Sr ions from the NE axoplasm as compared with that in the presence of Ca ions.

Topography and Affinity of Calcium Sensors of Exo- and Endocytosis in Motor Nerve Terminals

Measurements with extracellular microelectrode technique showed that depolarization of motor nerve terminals in frog cutaneous pectoris muscle with high-potassium solution (40 mM K(+)) increased frequency of miniature end-plate currents. Both fast intracellular calcium chelator BAPTA-AM and slow chelator EGTA-AM equally moderated the increase in the frequency of miniature end-plate currents. Intravital fluorescent microscopy with FM 1-43 dye showed that under conditions of stimulation of neurotransmitter exocytosis and secretion with high-potassium solution, internalization of the dye into newly-formed endocytotic synaptic vesicles proceeded both in the control and in the presence of EGTA-AM. In contrast, internalization of the dye was not observed in the presence of BAPTA-AM. It was concluded that asynchronous exocytosis of synaptic vesicles goes on in the active zones enriched with Ca-channels due to activation of high-affinity Ca-site in Ca-macrodomain. Endocytosis of vesicles is probably initiated by Ca-microdomain during activation of low-affinity Ca-site in the immediate proximity to the Ca channel.

Material basis for inhibition of Dragon’s Blood on evoked discharges of wide dynamic range neurons in spinal dorsal horn of rats

In vivo experiments were designed to verify the analgesic effect of Dragon's Blood and the material basis for this effect. Extracellular microelectrode recordings were used to observe the effects of Dragon's Blood and various combinations of the three components (cochinchinenin A, cochinchinenin B, and loureirin B) extracted from Dragon's Blood on the discharge activities of wide dynamic range (WDR) neurons in spinal dorsal horn (SDH) of intact male Wistar rats evoked by electric stimulation at sciatic nerve. When the Hill's coefficients describing the dose-response relations of drugs were different, based on the concept of dose equivalence, the equations of additivity surfaces which can be applied to assess the interaction between three drugs were derived. Adopting the equations and Tallarida's isobole equations used to assess the interaction between two drugs with dissimilar dose-response relations, the effects produced by various combinations of the three components in modulating the evoked discharge activities of WDR neurons were evaluated. Results showed that Dragon's Blood and its three components could inhibit the evoked discharge frequencies of WDR neurons in a concentration-dependent way. The Hill's coefficients describing dose-response relations of three components were different. Only the combined effect of cochinchinenin A, cochinchinenin B and loureirin B was similar to that of Dragons Blood. Furthermore, the combined effect was synergistic. This investigation demonstrated that through the synergistic interaction of the three components Dragon's Blood could interfere with the transmission and processing of pain signals in spinal dorsal horn. All these further proved that the combination of cochinchinenin A, cochinchinenin B, and loureirin B was the material basis for the analgesic effect of Dragon's Blood.

Localizing Cortical Computations during Visual Selection

Local field potentials (LFPs) and spikes are two signals that can be recorded from the brain using extracellular microelectrodes. A study by Monosov et al. in this issue of Neuron using timing relations between these signals suggests that selection of a target from an array of distractors is a computational operation performed specifically and locally in the frontal eye field (FEF).

Fatty acids modulate transmitter release and functioning of potassium channels in motor nerve endings

An extracellular microelectrode applied to the neuromuscular junction of the frog sternocutaneus muscle was used to study effects of saturated (myristic and arachidic) and unsaturated (arachidonic and oleic) fatty acids on transmitter release and potassium currents in a nerve ending. All these fatty acids decreased evoked transmitter release. Unsaturated fatty acids decreased the amplitude of voltage-dependent and calciumactivated potassium currents in a nerve ending, whereas saturated acids were ineffective. Rhythmic stimulation applied in the presence of arachidonic and oleic acids induced more pronounced facilitation of the transmitter release, whereas the effects of myristic and arachidic acids did not differ from control values. Deoxycholate was not able to reproduce the effects of fatty acids on the transmitter release and potassium current in a nerve ending. It was concluded that fatty acids can modulate transmitter release and synaptic excitatory transmission; these effects can have a significant influence on molecular mechanisms of exocytosis of synaptic vesicles and electrogenesis in a motor nerve ending. The latter effect is characteristic of unsaturated (arachidonic and oleic) acids, whereas the former effect does not depend on the degree of unsaturation at all.

Modulation of Neurotransmitter Release by Carbon Monoxide at the Frog Neuro-Muscular Junction

Carbon monoxide (CO) is an endogenous gaseous messenger, which regulates numerous physiological functions in a wide variety of tissues. Using extracellular microelectrode recording from frog neuro-muscular preparation the mechanisms of exogenous and endogenous CO action on evoked quantal acetyl-choline (Ach) release were studied. It was shown that CO application increases Ach-release in dose-dependent manner without changes in pre-synaptic Na+ and K+ currents. The effect of exogenous CO on Ach-release was decreased by prior application of guanylate cyclase inhibitor ODQ and prevented by application of a cyclic guanylate monophosphate (cGMP) analog 8Br-cGMP. Pre-treatment of the preparation with adenylate cyclase inhibitor MDL-12330A has completely abolished the effect of CO, whereas elevation of intracellular level of cyclic adenosine monophosphate (cAMP) mimicked and eliminated CO action. Application of cGMP-activated phosphodiesterase-2 inhibitor EHNA did not prevent CO action, whereas inhibition of cGMP-inhibited phosphodiesterase-3 by quazinone has partially blocked the effect of CO. Utilizing immuno-histochemical methods CO-producing enzyme heme-oxygenase-2 (HO-2) was shown to be expressed in skeletal muscle fibers, mostly in sub-sarcolemmal region, karyolemma and sarcoplasmic reticulum. Zn-protoporphirin-IX, the selective HO-2 blocker, has depressed Ach-release, suggesting the tonic activating effect of endogenous CO on pre-synaptic function. These results suggest that facilitatory effect of CO on Ach-release is mediated by elevation of intracellular cAMP level due to activation of adenylate cyclase and decrease of cAMP breakdown. As such, endogenous skeletal muscle-derived CO mediates tonic retrograde up-regulation of neuro-transmitter release at the frog neuro-muscular junction.

Spontaneous Retinal Activity Mediates Development of Ocular Dominance Columns and Binocular Receptive Fields in V1

The mechanisms that give rise to ocular dominance columns (ODCs) during development are controversial. Early experiments indicated a key role for retinal activity in ODC formation. However, later studies showed that in those early experiments, the retinal activity perturbation was initiated after ODCs had already formed. Moreover, recent studies concluded that early eye removals do not impact ODC segregation. Here we blocked spontaneous retinal activity during the very early stages of ODC development. This permanently disrupted the anatomical organization of ODCs and led to a dramatic increase in receptive field size for binocular cells in primary visual cortex. Our data suggest that early spontaneous retinal activity conveys crucial information about whether thalamocortical axons represent one or the other eye and that this activity mediates binocular competition important for shaping receptive fields in primary visual cortex.

Effects of electrical stimulation of ventral septal area on firing rates of pyrogen-treated thermosensitive neurons in preoptic anterior hypothalamus from rabbits

Although there is considerable evidence supporting that fever evolved as a host defense response, it is important that the rise in body temperature would not be too high. Many endogenous cryogens or antipyretics that limit the rise in body temperature have been identified. Endogenous antipyretics attenuate fever by influencing the thermoregulatory neurons in the preoptic anterior hypothalamus (POAH) and in adjacent septal areas including ventral septal area (VSA). Our previous study showed that intracerebroventricular (I.C.V.) injection of interleukin-1 β (IL-1 β) affected electrophysiological activities of thermosensitive neurons in VSA regions, and electrical stimulation of POAH reversed the effect of IL-1 β. To further investigate the functional electrophysiological connection between POAH and VSA and its mechanisms in thermoregulation, the firing rates of thermosensitive neurons in POAH of forty-seven unit discharge were recorded by using extracellular microelectrode technique in New Zealand white rabbits. Our results show that the firing rates of the warm-sensitive neurons decreased significantly and those of the cold-sensitive neurons increased in POAH when the pyrogen (IL-1 β) was injected I.C.V. The effects of IL-1 β on firing rates in thermosensitive neurons of POAH were reversed by electrical stimulation of VSA. An arginine vasopressin (AVP) V1 antagonist abolished the regulatory effects of VSA on the firing rates in thermosensitive neurons of POAH evoked by IL-1 β. However, an AVP V2 antagonist had no effects. These data indicated that VSA regulates the activities of the thermosensitive neurons of POAH through AVP V1 but not AVP V2 receptor.

Activation of group II metabotropic glutamate receptors reduces directional selectivity in retinal ganglion cells

In the mammalian retina, high levels of the group II metabotropic glutamate receptor (mGluR) subtype are expressed in starburst amacrine cells. A prominent role of starburst amacrine cells is the generation of directional selectivity in ON–OFF directionally selective retinal ganglion cells (DS RGCs). Extracellular microelectrodes were used to study the effects of activation of group II mGluRs on the responses of rabbit ON–OFF DS RGCs to a moving light stimulus. Directionally selective responses in these RGCs were substantially reduced by the selective group II mGluR agonist DCG-IV. DCG-IV brought out a response to movement in the null direction that was similar in magnitude and time course to the response to movement in the preferred direction. This effect of DCG-IV was reversed by the group II mGluR antagonist EGLU. Application of EGLU alone failed to alter directional selectivity in the RGCs but did reduce the response to movement in the preferred direction. To determine whether group II mGluRs modulate the release of the neurotransmitter acetylcholine from starburst amacrine cells, the effect of DCG-IV on ON–OFF DS RGCs was examined in the presence of ambenonium, an acetylcholinesterase inhibitor. When applied alone, ambenonium greatly prolonged the responses of ON–OFF DS RGCs to a light stimulus. This effect of ambenonium was completely abolished upon application of DCG-IV. Overall, the results suggest that postsynaptic group II mGluRs have the potential to influence directional selectivity in RGCs by inhibiting transmitter release from starburst amacrine cells.

An Automatic Algorithm for Stationary Segmentation of Extracellular Microelectrode Recordings

Extracellular microelectrode recordings (MER) often contain artifact from a variety of sources that confound traditional signal-processing techniques that require stationary signal segments. We designed an algorithm to locate the longest stationary segment of MER signals. In this paper we provide a description of the segmentation algorithm and its performance assessment. Simulation results demonstrate that the automatic segmentation algorithm we proposed is capable of accurately identifying the boundaries of the longest stationary segments in MER signals. In our simulation study the segmentation algorithm correctly identified the boundaries of the longest MER stationary segments in 99.5% of the cases.

Activation of cat SII cortex by flutter stimulation of contralateral vs. ipsilateral forepaws

A distinguishing feature of SII cortex is that it receives substantial input from skin mechanoreceptors located on both sides of the body. It remains uncertain, however, if integration of bilateral inputs occurs mainly in those regions of SII that represent near-midline body regions or also occurs to a significant extent in those regions of SII that represent the distal extremities. This issue was addressed using extracellular microelectrode recordings in cat SII in combination with the method of optical intrinsic signal (OIS) imaging. Stimulation of the central pad of either the contra- or ipsilateral forepaw with a 25-Hz sinusoidal vertical skin displacement (“skin flutter”) stimulus evoked a prominent OIS response (“activation”) in an extensive anteroposterior sector of SII. In the anteriorly located SII region that yielded the maximal OIS response to stimulation of the contralateral central pad, neurons consistently possessed receptive fields that included the stimulated skin site. This “forepaw” SII region also exhibited significant although 75% weaker OIS activation in response to stimulation of the ipsilateral central pad. Stimulation of the central pads of either contra- or ipsilateral forepaws also evoked OIS activation in the posteriorly located ‘hindlimb’ region of SII—defined as the SII region comprised of neurons with receptive fields on the contralateral hindlimb. The OIS response to ipsilateral central pad stimulation was strongest in the posterior SII region that borders the suprasylvian fringe—a region in which neurons have very large, and frequently bilateral, cutaneous receptive fields. The results indicate that widespread regions within cat SII receive cutaneous inputs from the ipsilateral distal forelimb. It is suggested that the functional role of these ipsilateral inputs may be different in different SII regions.

Nocifensive reflex-related on- and off-cells in the pedunculopontine tegmental nucleus, cuneiform nucleus, and lateral dorsal tegmental nucleus

Cholinergic projections from the pedunculopontine tegmental nucleus (PPTg) to the rostral ventromedial medulla (RVM) have been implicated in nociceptive modulation. The goal of this study was to identify neurons with nocifensive reflex-related activity in the mesopontine tegmentum including the PPTg. This study used the same behavioral neurophysiological classification system to identify neurons as has been extensively described in the RVM. Extracellular microelectrode recording was conducted in lightly anesthetized rats. Changes in firing associated with the noxious heat-evoked tail flick reflex were used to classify neurons as “on-cells” (displayed a burst in neuronal activity associated with the reflex), “off-cells” (displayed a pause in activity), and neutral cells (showed no response). Of 188 neurons studied in 23 rats, 77 were classified as on-cells, 14 as off-cells, the remainder as neutral cells. Recordings during periods without noxious stimulation found that some of the on- and off-cells displayed spontaneous transitions between active and silent periods termed cell cycling. The distribution of on- and off-cells in the mesopontine tegmentum overlapped and included the cholinergic PPTg and lateral dorsal tegmental nucleus identified by NADPH diaphorase staining, as well as the cuneiform nucleus and periaqueductal gray. The mesopontine tegmentum thus contains nocifensive reflex-related neurons with neurophysiological characteristics similar to those reported in the RVM. Neurons showing reflex-related activity were frequently encountered in the cholinergic PPTg and LDTg. Further studies will be required to determine whether these neurons modulate nociception through a link to the RVM.

Spatiotemporal receptive field properties of epiretinally recorded spikes and local electroretinograms in cats

BackgroundReceptive fields of retinal neural signals of different origin can be determined from extracellular microelectrode recordings at the inner retinal surface. However, locations and types of neural processes generating the different signal components are difficult to separate and identify. We here report epiretinal receptive fields (RFs) from simultaneously recorded spikes and local electroretinograms (LERGs) using a semi-chronic multi-electrode in vivo recording technique in cats. Broadband recordings were filtered to yield LERG and multi unit as well as single unit spike signals. RFs were calculated from responses to multifocal pseudo-random spatiotemporal visual stimuli registered at the retinal surface by a 7-electrode array.ResultsLERGs exhibit spatially unimodal RFs always centered at the location of the electrode tip. Spike-RFs are either congruent with LERG-RFs (N = 26/61) or shifted distally (N = 35/61) but never proximally with respect to the optic disk. LERG-RFs appear at shorter latencies (11.9 ms ± 0.5 ms, N = 18) than those of spikes (18.6 ms ± 0.4 ms, N = 53). Furthermore, OFF-center spike-RFs precede and have shorter response rise times than ON-center spike-RFs. Our results indicate that displaced spike-RFs result from action potentials of ganglion cell axons passing the recording electrode en route to the optic disk while LERG-RFs are related to superimposed postsynaptic potentials of cells near the electrode tip.ConclusionBesides contributing to the understanding of retinal function we demonstrate the caveats that come with recordings from the retinal surface, i.e., the likelihood of recordings from mixed sets of retinal neurons. Implications for the design of an epiretinal visual implant are discussed.

Excitation of Medullary Respiratory Neurons in REM Sleep

To study tonic inputs to medullary respiratory neurons during rapid eye movement (REM) sleep. Single medullary-respiratory-neuron recordings during sleep with spontaneous breathing and during apnea caused by mechanical hyperventilation. Academic laboratory. Three tracheostomized adult cats implanted for polysomnography and extracellular microelectrode recordings. Single medullary-respiratory-neuron recordings during spontaneous breathing and mechanical hyperventilation to apnea during non-REM (NREM) and REM sleep. Most but not all respiratory cells of all types (pre-inspiratory, decrementing, augmenting and late inspiratory, phase-spanning, and expiratory) were more active in REM sleep than in NREM sleep during both spontaneous breathing and apnea induced by mechanical hyperventilation. The mean discharge rate of the cells during spontaneous breathing in NREM sleep was 16.7 impulses per second and in REM sleep was 26.5 impulses per second. During ventilator-induced apnea, the mean rates were 10 impulses per second in NREM sleep and 17.5 per second during REM sleep. The increase in activity in REM sleep occurred after a delay of several seconds from the onset of REM sleep. Respiratory cells were excited at times individually and at other times simultaneously in either a reciprocal or nonreciprocal pattern. The degree of excitation of a neuron in REM sleep during ventilator-induced apnea was proportional to the degree of excitation of the neuron in REM sleep during spontaneous breathing. Medullary respiratory neurons are excited individually and collectively in REM sleep. The excitation occurs with a delay after the onset of the state and can stimulate and/or disorganize breathing.

Thresholds for Activation of Rabbit Retinal Ganglion Cells with Relatively Large, Extracellular Microelectrodes

To investigate the responses of retinal ganglion cells (RGCs) to electrical stimulation, using electrodes comparable in size to those used in human studies investigating the feasibility of an electronic retinal prosthesis. Rabbit retinas were stimulated in vitro with current pulses applied to the inner surface with 125- and 500-mum diameter electrodes while the responses of RGCs were recorded extracellularly. Both short-latency (SL; 3-5 ms) and long-latency (LL; >/=9 ms) responses were observed after electrical stimulation within the receptive field of an RGC. With short, 0.1-ms current pulses, the threshold current for the SL cell response was significantly lower than that for the LL cell response. With long (10- to 20-ms) pulses, the threshold currents for the SL and LL cell responses were very similar. The threshold current for the SL cell response increased more steeply than did the LL cell response when the electrode was displaced from the point of lowest electrical threshold, either above or along the surface of the retina. Stimulation of an RGC axon outside of the cell's receptive field produced only an SL response. For 0.1-ms duration pulses, the threshold current for the axonal response was significantly higher than the threshold current for the SL cell response. At pulse durations > 1 ms, the thresholds were very similar. RGC responses to electrical stimulation depend on the current pulse duration and location of the stimulating electrode. For an epiretinal prosthesis, short-duration current pulses may be preferable since they result in a more localized activation of the retina.

Adenosine A1 receptors modulate the discharge activities of inspiratory and biphasic expiratory neurons in the medial region of Nucleus Retrofacialis of neonatal rat in vitro

This study investigated whether adenosine A1 receptors could modulate respiratory rhythm in mammals. Experiments were performed in in vitro brainstem slice preparations from neonatal rats. These preparations included the medial region of Nucleus Retrofacialis (mNRF) with the hypoglossal nerve (XII nerve) rootlets retained. The rhythmical discharges of the biphasic expiratory (biphasic E) neurons/inspiratory neurons (I neurons) and activities of the XII nerve rootlets were simultaneously recorded by using extracellular microelectrodes and suction electrodes, respectively. Roles of adenosine A1 receptors in modulation of respiratory rhythm were investigated by administration of the adenosine A1 receptor agonist R-phenylisopropyl-adenosine (R-PIA, 10 μM) and its specific antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 2 μM). DPCPX decreased the respiratory period (27.19%) and expiratory duration (28.27%) of biphasic E neurons and at the same time increased the peak discharge frequency (48.13%). By contrast, R-PIA produced opposite effects. On the other hand, the effects of DPCPX and R-PIA on the I neurons were similar to that on the biphasic E neurons except that R-PIA shortened the discharge duration of I neurons (34.12%) and decreased the peak discharge frequency (37.75%) in the middle phase of inspiration, but not in the initial and terminal phases. These results suggest that adenosine A1 receptors are involved in the phase-switching between expiration and inspiration by affecting biphasic E neurons. Activation of adenosine A1 receptors may modulate the inhibitory synaptic inputs from I neurons to biphasic E neurons.

Calcium dependence of uni-quantal release latencies and quantal content at mouse neuromuscular junction

Uni-quantal endplate currents (EPC) were recorded at mouse diaphragm neuromuscular synapse by extracellular microelectrode during motor nerve stimulation. The probability of release expressed as quantal content m(o), and variability of synaptic latencies expressed as P90 were estimated in the presence of extracellular calcium ([Ca2+]o) varying between 0.2 and 0.6 mM in the bathing solution. At 0.2 mM ([Ca2+]o), m(o) was low (0.10) and many of long-latency EPCs were present during the late phase of the release (P90 = 2.44 ms). No change in m(o) was found when ([Ca2+]o) was 0.3 mM, but P90 decreased by 39 %. For latency shortening, saturating concentration of ([Ca2+]o) was 0.4 mM, when P90 was 1.49 ms and latencies did not further change at 0.5 and 0.6 mM ([Ca2+]o). In the latter concentrations, however, an increase of m(o) was still observed. It can be concluded that the early phase of the secretion did not significantly change when ([Ca2+]o) was raised and that only the late phase of the release depends on extracellular calcium up to 0.4 mM.

Functional imaging of the embryonic pacemaking and cardiac conduction system over the past 150 years: technologies to overcome the challenges.

Early analyses of cardiac pacemaking and conduction system (CPCS) development relied on classic histology and visual inspection of the beating heart. Current techniques that facilitate delineation of the CPCS include the use of specific antibody markers and transgenic mouse lines specifically expressing reporter genes. Assaying the function of tiny embryonic hearts required an increase in the level of spatial and temporal resolution. Current methods for such analyses include the use of intracellular and extracellular microelectrodes, echocardiography, rapid optical imaging using fluorescent dyes, and most recently optical coherence tomography. This review will focus on methods developed to investigate the functional emergence of the embryonic cardiac conduction system. Where appropriate, the methods used to delineate the anatomic pathways will also be discussed. The combination of techniques to capture both morphological and functional data from the CPCS will further improve with continued interdisciplinary collaboration. The Supplementary Material referred to in this article can be found at the Anatomical Record website (http://www.interscience.wiley.com/jpages/0003-276X/suppmat).

Nociceptive excited and inhibited neurons within the pedunculopontine tegmental nucleus and cuneiform nucleus

The pedunculopontine tegmental nucleus (PPTg) is defined by its collection of cholinergic neurons surrounding the lateral portion of the superior cerebellar peduncle at the midbrain pontine junction. Antinociceptive functions have been attributed to the PPTg since electrical stimulation as well as injection of cholinergic agonists in this area produces analgesia. Nociceptive neurons have also been reported in the vicinity of the PPTg and cuneiform nucleus (CN). However, specific histochemical localization of nociceptive modulatory neurons has not been determined. Thus, the goal of this study was to classify neurons according to their response to a noxious stimulus and map their location based on staining of the cholinergic neurons in the PPTg. Extracellular microelectrode recordings were conducted in 19 male Sprague–Dawley rats under light halothane anesthesia. For each neuron identified, a series of noxious tail pinches were administered. The electrode tracts were marked with ionophoresis of pontamine blue. The location of 112 recorded neurons was determined on sections stained with NADPH diaphorase to identify the cholinergic boundaries of the PPTg. Neurons were classified into one of three cell types based on their consistent response to a noxious tail pinch (excited, inhibited, and non-responsive). Tail pinch excited neurons ( n=16), inhibited neurons ( n=10) and non-responsive neurons ( n=23) were mapped within the cholinergic boundaries of the PPTg. Excited ( n=9), inhibited ( n=10) and non-responsive neurons ( n=10) were also found more dorsally within the cuneiform nucleus. Thus, this study localizes nociception-responsive neurons to the region of the largely cholinergic PPTg, as well as the noncholinergic cuneiform nucleus.