Distribution of melanin-concentrating hormone neurons projecting to the medial mammillary nucleus

The melanin-concentrating hormone and neuropeptide glutamic acid-isoleucine are expressed in neurons located mainly in the hypothalamus that project widely throughout the CNS. One of the melanin-concentrating hormone main targets is the medial mammillary nucleus, but the exact origin of these fibers is unknown. We observed melanin-concentrating hormone and neuropeptide glutamic acid-isoleucine immunoreactive fibers coursing throughout the mammillary complex, showing higher density in the pars lateralis of the medial mammillary nucleus, while the lateral mammillary nucleus showed sparse melanin-concentrating hormone innervation. The origins of these afferents were determined by using implant of the retrograde tracer True Blue in the medial mammillary nucleus. Double-labeled neurons were observed in the lateral hypothalamic area, rostromedial zona incerta and dorsal tuberomammillary nucleus. A considerable population of retrogradely labeled melanin-concentrating hormone perikaryal profiles was also immunoreactive to neuropeptide glutamic acid-isoleucine (74+/-15% to 85+/-15%). The afferents from the lateral hypothalamic area, rostromedial zona incerta and dorsal tuberomammillary nucleus to the medial mammillary nucleus were confirmed using implant of the anterograde tracer Phaseolus vulgaris leucoagglutinin. In addition, using double-labeled immunohistochemistry, we found no co-localization between neurons expressing melanin-concentrating hormone and adenosine deaminase (histaminergic marker) in the dorsal tuberomammillary nucleus. We hypothesize that these melanin-concentrating hormone projections participate in spatial memory process mediated by the medial mammillary nucleus. These pathways would enable the animal to look for food during the initial moments of appetite stimulation.     

Attenuated glycine receptor function reduces excitability of mouse medial vestibular nucleus neurons

Spontaneous activity in medial vestibular nucleus (MVN) neurons is modulated by synaptic inputs. These inputs are crucial for maintaining gaze and posture and contribute to vestibular compensation after lesions of peripheral vestibular organs. We investigated how chronically attenuated glycinergic input affects excitability of MVN neurons. To this end we used three mouse strains (spastic, spasmodic, and oscillator), with well-characterized naturally occurring mutations in the inhibitory glycine receptor (GlyR). First, using whole-cell patch-clamp recordings, we demonstrated that the amplitude of the response to rapidly applied glycine was dramatically reduced by 25 to 90% in MVN neurons from mutant mice. We next determined how reduced GlyR function affected MVN neuron output. Neurons were classified using two schemas: (1) the shape of their action potential afterhyperpolarization (AHP); and (2) responses to hyperpolarizing current injection. In the first schema, neurons were classified as types A, B and C. The prevalence of type C neurons in the mutant strains was significantly increased. In the second schema, the proportion of neurons lacking post inhibitory rebound firing (PRF-deficient) was increased. In both schemas an increase in AHP amplitude was a common feature of the augmented neuron group (type C, PRF-deficient) in the mutant strains. We suggest increased AHP amplitude reduces overall excitability in the MVN and thus maintains network function in an environment of reduced glycinergic input.     

Characteristics of near response cells projecting to the oculomotor nucleus.

1. Previous work has shown neurons just dorsal and lateral to the oculomotor nucleus that increase their firing rate with increases in the angle of ocular convergence. It has been suggested that the output of these midbrain near response cells might provide the vergence command needed by the medial rectus motoneurons. However, lens accommodation ordinarily accompanies convergence, and a subsequent study showed that only about one-half of these midbrain near response cells carried a signal related exclusively to vergence. One hypothesis suggested by this finding is that this subgroup of neurons might have a unique role in providing a "pure" vergence signal to the medial rectus motoneurons. 2. In the present study extracellular recordings were made from midbrain near response cells in monkeys while eye position and lens accommodation were measured. The monkeys viewed targets through an optical system that allowed the accommodative and ocular vergence demands to be manipulated independently. This approach was used to produce a partial dissociation of accommodative and vergence responses, so that an accommodative and vergence coefficient could be determined for each cell, by the use of the following equation FR = R0 + kda x AR + kdv x CR where FR is the firing rate of the near response cell, R0 is the predicted firing rate for a distant target, kda is the (dissociated) accommodation coefficient, AR is the accommodative response, kdv is the (dissociated) vergence coefficient, and CR is the convergence response. 3. The vergence and accommodation coefficients were determined for a large number of midbrain near response cells, including a subset that could be antidromically activated from the medial rectus subdivisions of the oculomotor nucleus. Some near response neurons were found with signals related exclusively to convergence (i.e., kdv greater than 0 and kda = 0), whereas several others had signals related exclusively to lens accommodation (i.e., kda greater than 0 and kdv = 0). The majority of the near response cells had signals related to both responses (i.e., kda not equal to 0 and kdv not equal to 0). Furthermore, the vergence and accommodation coefficients of near response cells appeared to be continuously distributed. Some cells had negative accommodation or vergence coefficients. 4. The 17 near response cells that could be antidromically activated from the oculomotor nucleus presumably provide vergence signals to the medial rectus motoneurons. Although all had positive vergence coefficients, only four of these cells carried signals that were related exclusively to vergence.(ABSTRACT TRUNCATED AT 400 WORDS)     

Inhibitory synaptic transmission differs in mouse type A and B medial vestibular nucleus neurons in vitro

Fast inhibitory synaptic transmission in the medial vestibular nucleus (MVN) is mediated by GABA(A) receptors (GABA(A)Rs) and glycine receptors (GlyRs). To assess their relative contribution to inhibition in the MVN, we recorded miniature inhibitory postsynaptic currents (mIPSCs) in physiologically characterized type A and type B MVN neurons. Transverse brain stem slices were prepared from mice (3-8 wk old), and whole cell patch-clamp recordings were obtained from visualized MVN neurons (CsCl internal; Vm = -70 mV; 23 degrees C). In 81 MVN neurons, 69% received exclusively GABA(A)ergic inputs, 6% exclusively glycinergic inputs, and 25% received both types of mIPSCs. The mean amplitude of GABA(A)R-mediated mIPSCs was smaller than those mediated by GlyRs (22.6 +/- 1.8 vs. 35.3 +/- 5.3 pA). The rise time and decay time constants of GABA(A)R- versus GlyR-mediated mIPSCs were slower (1.3 +/- 0.1 vs. 0.9 +/- 0.1 ms and 10.5 +/- 0.3 vs. 4.7 +/- 0.3 ms, respectively). Comparison of type A (n = 20) and type B (n = 32) neurons showed that type A neurons received almost exclusively GABA(A)ergic inhibitory inputs, whereas type B neurons received GABA(A)ergic inputs, glycinergic inputs, or both. Intracellular labeling in a subset of MVN neurons showed that morphology was not related to a MVN neuron's inhibitory profile (n = 15), or whether it was classified as type A or B (n = 29). Together, these findings indicate that both GABA and glycine contribute to inhibitory synaptic processing in MVN neurons, although GABA dominates and there is a difference in the distribution of GABA(A) and Gly receptors between type A and type B MVN neurons.     

Properties of neurons from the rat medial vestibular nucleus in microexplant culture

This study is a first step in an attempt to identify the factors which determine and maintain the electrophysiological phenotype(s) of mature neurons of the medial vestibular nucleus (MVN). We cultured MVN microexplants obtained from slices of the brainstem of newborn rats, using a hollow punching needle. The electrophysiological maturation of the neurons was followed by analyzing their responses to 1 s steps of current of increasing amplitude. The maximal number of spikes that was generated in response to such stimuli increased dramatically over time in vitro. However, even after 28 days in vitro, it did not exceed about 60 spikes/s. At this stage of culture, the input-output properties of the spike generator of the MVN neurons were similar to those observed in brainstem slices of newborn rats, but clearly inferior to those of adult neurons which can generate sustained firing up to 150-200 spikes/s.     

Antidromic identification of midbrain near response cells projecting to the oculomotor nucleus

Summary Medial rectus motoneurons carry both conjugate and vergence eye position signals. Abducens internuclear neurons, whose axons travel in the medial longitudinal fasciculus, provide these motoneurons with the major signal for conjugate eye movements but not for vergence eye movements. A vergence signal appropriate for these motoneurons is seen on the near response cells that are found in the mesencephalic reticular formation within 2 mm of the oculomotor nucleus. The goal of the present study was to determine if midbrain near response cells project to the medial rectus subdivision of the oculomotor nucleus. Near response cells were recorded in two trained rhesus monkeys with ocular search coils. A stimulating electrode was positioned within the medial rectus subdivision of the oculomotor nucleus. Twenty-eight near response cells were found that could be driven by single pulse microstimulation of the ipsilateral medial rectus subdivision. In all cases, antidromic activation was confirmed by collision testing. Attempts to antidromically activate midbrain near response cells from the contralateral medial rectus subdivision were unsuccessful. Most antidromically activated cells had a steady state firing rate proportional to vergence angle. One cell also showed burst activity during the vergence eye movements. Divergence cells were not antidromically activated.     

Long-term plasticity of ipsilesional medial vestibular nucleus neurons after unilateral labyrinthectomy

Unilateral labyrinthectomy results in oculomotor and postural disturbances that regress in a few days during vestibular compensation. The long-term (after 1 mo) consequences of unilateral labyrinthectomy were investigated by characterizing the static and dynamic membrane properties of the ipsilesional vestibular neurons recorded intracellularly in guinea pig brain stem slices. We compared the responses of type A and type B medial vestibular nucleus neurons identified in vitro to current steps and ramps and to sinusoidal currents of various frequencies. All ipsilesional vestibular neurons were depolarized by 6-10 mV at rest compared with the cells recorded from control slices. Both their average membrane potential and firing threshold were more depolarized, which suggests that changes in active conductances compensated for the loss of excitatory afferents. The afterhyperpolarization and discharge regularity of type B but not type A neurons were increased. All ipsilesional vestibular cells became more sensitive to current injections over a large range of frequencies (0.2-30 Hz), but this increase in sensitivity was greater for type B than for type A neurons. This was associated with an increase of the peak frequency of linear response restricted to type B neurons, from 4-6 to 12-14 Hz. Altogether, we show that long-term vestibular compensation involves major changes in the membrane properties of vestibular neurons on the deafferented side. Many of the static and dynamic membrane properties of type B neurons became more similar to those of type A neurons than in control slices, leading to an increase in the overall homogeneity of medial vestibular nucleus neurons.     

Postnatal development of spike generation in rat medial vestibular nucleus neurons

Image stability during self motion depends on the combined actions of the vestibuloocular and optokinetic reflexes (VOR and OKR, respectively). Neurons in the medial vestibular nucleus (MVN) participate in the VOR and OKR by firing in response to both head and image motion. Their intrinsic spike-generating properties enable MVN neurons to modulate firing rates linearly over a broad range of input amplitudes and frequencies such as those that occur during natural head and image motion. This study examines the postnatal development of the intrinsic spike-generating properties of rat MVN neurons with respect to maturation of peripheral vestibular and visual function. Spike generation was studied in a brain stem slice preparation by recording firing responses to current injected intracellularly through whole cell patch electrodes. MVN neurons fired spontaneously and modulated their firing rate in response to injected current at all postnatal ages. However, the input-output properties of the spike generator changed dramatically during the first two postnatal weeks. Neurons younger than postnatal day 10 could not fire faster than 80 spikes/s, modulated their firing rates over a limited range of input amplitudes, and tended to exhibit a nonlinear relationship between input current and mean evoked firing rate. In response to sustained depolarization, firing rates declined significantly in young neurons. Response gains tended to be highest in the first few postnatal days but varied widely across neurons and were not correlated with age. By about the beginning of the third postnatal week, MVN neurons could fire faster than 100 spikes/s in response to a broad range of input amplitudes, exhibited predominantly linear current-firing rate relationships, and adapted little in response to sustained depolarization. Concomitant decreases in action potential width and the time course of the afterhyperpolarization suggest that changes in potassium currents contribute to the maturation of the MVN neuronal spike generator. The results demonstrate that developmental changes in intrinsic membrane properties enable MVN neurons to fire linearly in response to a broad range of stimuli in time for the onset of visual function at the beginning of the third postnatal week.     

Histamine depolarizes rat medial vestibular nucleus neurons recorded intracellularly in vitro

The effects of histamine (HA) on the resting membrane potential and input resistance of rat medial vestibular nucleus (MVN) neurons were investigated using intracellular recording techniques from a submerged brain slice preparation. The exogenous application of HA predominantly produced a concentration-dependent membrane depolarization and induction of action potential firing. The depolarization exhibited a rapid onset, a slow recovery, and usually occurred in the absence of any apparent change in conductance. These effects of HA could be mimicked by the H2-agonist impromidine and were reversibly blocked by the H2-antagonist cimetidine. Tetrodotoxin (TTX) or low calcium/high magnesium-containing media failed to block completely the HA-induced depolarization supporting a direct postsynaptic receptor mediated action of HA. The diminished HA-induced depolarization observed following pretreatment with TTX cannot exclude an additional presynaptic action by HA. The present findings reveal that HA exerts a novel direct excitation of rat MVN neurons through an H2-receptor.     

Physiological properties of sensory trigeminal neurons projecting to mesencephalic parabrachial area in the cat

1. One hundred forty-one trigeminomesencephalic neurons in the sensory trigeminal nucleus of cats anesthetized with alpha-chloralose were identified by antidromic stimulation of the mesencephalic parabrachial area (PBA) which includes the nucleus cuneiformis, lateral periaqueductal gray matter, and the region between the inferior colliculus and brachium conjunctivum. 2. Neurons were categorized based on their responses to non-noxious and noxious mechanical and heat stimuli delivered to their peripheral receptive fields (RFs) including skin, mucosa, guard hairs, vibrissae, cornea, and tooth pulps. They were classified into three types: 48 nociceptive-specific (NS) neurons which responded to heavy pressure and/or noxious mechanical stimuli, and/or noxious radiant heat; 19 wide dynamic range (WDR) neurons which had a graded response to light tactile stimuli, noxious pinch, and/or noxious radiant heat; and 36 low-threshold mechanoreceptive (LTM) neurons which responded maximally to innocuous tactile stimuli. In subnucleus caudalis (Vc), NS and WDR neurons were the majority (75%) among the three types, while in the rostral subnuclei they were about one-half (54%) of the population. 3. The RFs were distributed over the orofacial and head region but mainly in the facial region. Twelve neurons (33% of the LTM neurons) responded to deflection of vibrissae and only 1 NS neuron out of 52 NS and WDR neurons tested responded to electrical stimulation of a tooth pulp. 4. A contralateral projection was dominant (57%), 30% projected ipsilaterally and 13% projected bilaterally.(ABSTRACT TRUNCATED AT 250 WORDS)     

Functional role of cyclic nucleotide-gated channels in rat medial vestibular nucleus neurons

Although cyclic nucleotide-gated (CNG) channels are expressed in numerous brain areas, little information is available on their functions in CNS neurons. The aim of the present study was to define the distribution of CNG channels in the rat medial vestibular nucleus (MVN) and their possible involvement in regulating MVN neuron (MVNn) excitability. The majority of MVNn expressed both CNG1 and CNG2 A subunits. In whole-cell current-clamp experiments carried out on brainstem slices containing the MVNn, the membrane-permeant analogues of cyclic nucleotides, 8-Br-cGMP and 8-Br-cAMP (1 m m ), induced membrane depolarizations (8.9 ± 0.8 and 9.2 ± 1.0 mV, respectively) that were protein kinase independent. The cGMP-induced depolarization was associated with a significant decrease in the membrane input resistance. The effects of cGMP on membrane potential were almost completely abolished by the CNG channel blockers, Cd²⁺ and l - cis -diltiazem, but they were unaffected by blockade of hyperpolarization-activated cyclic nucleotide-gated channels. In voltage-clamp experiments, 8-Br-cGMP induced non-inactivating inward currents (−22.2 ± 3.9 pA) with an estimated reversal potential near 0 mV, which were markedly inhibited by reduction of extracellular Na⁺ and Ca²⁺ concentrations. Membrane depolarization induced by CNG channel activation increased the firing rate of MVNn without changing the action potential shape. Collectively, these findings provide novel evidence that CNG channels affect membrane potential and excitability of MVNn. Such action should have a significant impact on the function of these neurons in sensory–motor integration processes. More generally, it might represent a broad mechanism for regulating the excitability of different CNS neurons.     

Resonance of spike discharge modulation in neurons of the guinea pig medial vestibular nucleus

The modulation of action potential discharge rates is an important aspect of neuronal information processing. In these experiments, we have attempted to determine how effectively spike discharge modulation reflects changes in the membrane potential in central vestibular neurons. We have measured how their spike discharge rate was modulated by various current inputs to obtain neuronal transfer functions. Differences in the modulation of spiking rates were observed between neurons with a single, prominent after hyperpolarization (AHP, type A neurons) and cells with more complex AHPs (type B neurons). The spike discharge modulation amplitudes increased with the frequency of the current stimulus, which was quantitatively described by a neuronal model that showed a resonance peak >10 Hz. Modeling of the resonance peak required two putative potassium conductances whose properties had to be markedly dependent on the level of the membrane potential. At low frequencies (< or =0.4 Hz), the gain or magnitude functions of type A and B discharge rates were similar relative to the current input. However, resting input resistances obtained from the ratio of the membrane potential and current were lower in type B compared with type A cells, presumably due to a higher level of active potassium conductances at rest. The lower input resistance of type B neurons was compensated by a twofold greater sensitivity of their firing rate to changes in membrane potential, which suggests that synaptic inputs on their dendritic processes would be more efficacious. This increased sensitivity is also reflected in a greater ability of type B neurons to synchronize with low-amplitude sinusoidal current inputs, and in addition, their responses to steep slope ramp stimulation are enhanced over the more linear behavior of type A neurons. This behavior suggests that the type B MVNn are moderately tuned active filters that promote high-frequency responses and that type A neurons are like low-pass filters that are well suited for the resting tonic activity of the vestibular system. However, the more sensitive and phasic type B neurons contribute to both low- and high-frequency control as well as signal detection and would amplify the contribution of both irregular and regular primary afferents at high frequencies.     

Adaptation of chicken vestibular nucleus neurons to unilateral vestibular ganglionectomy

Vestibular compensation refers to the behavioral recovery after a unilateral peripheral vestibular lesion. In chickens, posture and balance deficits are present immediately following unilateral vestibular ganglionectomy (UVG). After three days, most operated chickens begin to recover, but severe deficits persist in others. The tangential nucleus is a major avian vestibular nucleus whose principal cells are vestibular reflex projection neurons. From patch-clamp recordings on brain slices, the percentage of spontaneous spike firing principal cells, spike discharge rate, ionic conductances, and spontaneous excitatory postsynaptic currents (sEPSCs) were investigated one and three days after UVG. Already by one day after UVG, sEPSC frequency increased significantly on the lesion side, although no differences were detected in the percentage of spontaneous spike firing cells or discharge rate. In compensated chickens three days after UVG, the percentage of spontaneous spike firing cells increased on the lesion side and the discharge rate increased bilaterally. In uncompensated chickens three days after UVG, principal cells on the lesion side showed increased discharge rate and increased sEPSC frequency, whereas principal cells on the intact side were silent. Typically, silent principal cells exhibited smaller persistent sodium conductances and higher activation thresholds for the fast sodium channel than spiking cells. In addition, silent principal cells on the intact side of uncompensated chickens had larger dendrotoxin-sensitive potassium conductance, with a higher ratio of Kv1.1 surface/cytoplasmic expression. Increased sEPSC frequency in principal cells on the lesion side of uncompensated chickens was accompanied by decreased Kv1.2 immunolabeling of presynaptic terminals on principal cell bodies. Thus, both intrinsic ionic conductances and excitatory synaptic inputs play crucial roles at early stages after lesions. Unlike the principal cells in compensated chickens which showed similar percentages of spontaneous spike firing cells, discharge rates, and sEPSC frequencies bilaterally, principal cells in uncompensated chickens displayed gross asymmetry in these properties bilaterally.