Protracted development of responses to whisker deflection in rat trigeminal ganglion neurons

The rodent whisker-to-barrel pathway constitutes a major model system for studying experience-dependent brain development. Yet little is known about responses of neurons to whisker stimulation in young animals. Response properties of trigeminal ganglion (NV) neurons in 2-, 3-, and 4-week-old and adult rats were examined using extracellular single-unit recordings and controlled whisker stimuli. We found that the receptive field size of NV neurons is mature in 2-week-old animals while response latencies, magnitudes, and angular tuning continue to develop between 2 weeks of age and adulthood. At the earliest time recorded, NV neurons respond to stimulation of only one whisker and can be characterized as slowly or rapidly adapting (SA, RA). The proportion of SA and RA neurons remains constant during development. Consistent with known on-going myelination of NV axons, response latencies decrease with age, becoming adult-like during the third and fourth postnatal weeks for RA and SA neurons, respectively. Unexpectedly, we found that evoked response magnitudes increase several-fold during development becoming adult-like only during the fourth postnatal week. In addition, RA neurons become less selective for whisker deflection angle with age. Maturation of response magnitude and angular tuning is consistent with developmental changes in the mechanical properties of the whisker, the whisker follicle, and the surrounding tissues. The findings indicate that whisker-derived tactile inputs mature during the first postnatal month when whisker-related cortical circuits are susceptible to long-term modification by sensory experience. Thus normal developmental changes in sensory input may influence functional development of cortical circuits.     

Precise temporal responses in whisker trigeminal neurons

The ability of rats using their whiskers to perform fine tactile discrimination rivals that of humans using their fingertips. Rats must perform these discriminations rapidly and accurately while palpating the environment with their whiskers. This suggests that whisker-derived inputs produce a robust and reliable code, capable of capturing complex, high-frequency information. The first neural representation of whisker-derived stimulus information is in primary afferent neurons of the trigeminal ganglion. Here we demonstrate that there is a continuum of direction-dependent response profiles in trigeminal neurons and provide the first quantitative analysis of the encoding of complex stimuli by these neurons. We show that all classes of trigeminal ganglion neurons respond with highly reproducible temporal spike patterns to transient stimuli. Such a robust coding mechanism may allow rapid perception of complex tactile features.     

Responses of trigeminal ganglion neurons to the radial distance of contact during active vibrissal touch

Rats explore their environment by actively moving their whiskers. Recently, we described how object location in the horizontal (front-back) axis is encoded by first-order neurons in the trigeminal ganglion (TG) by spike timing. Here we show how TG neurons encode object location along the radial coordinate, i.e., from the snout outward. Using extracellular recordings from urethane-anesthetized rats and electrically induced whisking, we found that TG neurons encode radial distance primarily by the number of spikes fired. When an object was positioned closer to the whisker root, all touch-selective neurons recorded fired more spikes. Some of these cells responded exclusively to objects located near the base of whiskers, signaling proximal touch by an identity (labeled-line) code. A number of tonic touch-selective neurons also decreased delays from touch to the first spike and decreased interspike intervals for closer object positions. Information theory analysis revealed that near-certainty discrimination between two objects separated by 30% of the length of whiskers was possible for some single cells. However, encoding reliability was usually lower as a result of large trial-by-trial response variability. Our current findings, together with the identity coding suggested by anatomy for the vertical dimension and the temporal coding of the horizontal dimension, suggest that object location is encoded by separate neuronal variables along the three spatial dimensions: temporal for the horizontal, spatial for the vertical, and spike rate for the radial dimension.     

Preliminary studies on GABA-immunoreactive neurons in the rat trigeminal ganglion

GABA-immunoreactive (ir) primary sensory neurons have been reported in many studies. However, the role of GABA in the primary sensory neurons and their targets is quite open to question. The present study aimed to examine the GABA-ir neurons in the rat trigeminal ganglion (TG), for the first step of functional study on them. Some neurons in the TG showed GABA-ir, which were ranged from large to small size. The total number of examined TG neurons from 6 ganglia was 2,531, of which 462 neurons (18.3%) showed GABA-ir the large subpopulation of GABA-ir TG neurons is likely to involve in nerve-muscle functions, whereas medium and small subpopulations might participate in cutaneous nociceptive sensory function. The present findings demonstrated a considerable number of sensory neurons containing GABA in the rat TG. Functional studies on GABA-ir neurons in the TG would be an interesting and important issues in future studies. The next aim of our study is to examine the size distribution of GABAergic neurons and the coexistence with other neurotransmitters in the rat TG.     

Collateral projections of trigeminal ganglion neurons to both the principal sensory trigeminal and the spinal trigeminal nuclei in the rat

Employing a combination of fluorescent retro grade double labelling and immunofluorescence histo chemistry for substance P (SP) and calcitonin gene-relat ed peptide (CGRP), we examined collateral projections from single neurons in the trigeminal ganglion (TG) of the rat to both the principal sensory trigeminal nucleus (Vp) and the oral, interpolar or caudal subnuclei of the spinal trigeminal nucleus (Vo, Vi or Vc). In the rats that were unilaterally injected with fast blue (FB) into the Vp and with diamidino yellow (DY) into the Vo, Vi or Vc, neurons labelled with FB and/or DY were observed in the TG ipsilateral to the injections. Of the labelled TG neurons, about 2% were double labelled with both trac ers in the rats that were injected with FB into the Vp and with DY into the Vo or Vi, and about 10% were double labelled in the rats that were injected with FB into the Vp and with DY into the Vc. The results indicate that TG neurons sending their axons to the Vp project, by way of axon collaterals, to the Vc more frequently than to the Vo or Vi.Some of the TG neurons double labelled with FB and DY exhibited SP-or CGRP-like immunoreactivity (LI): Of the TG neurons that were double labelled with FB injected into the Vp and with DY injected into the Vo, Vi or Vc, about 38%, 49% and 42%, respectively, displayed SP-LI, and about 54%, 58% and 59%, respectively, showed CGRP-LI. Some of the SP-or CGRP-LI TG neurons that were double labelled with FB and DY were assumed to mediate pain signals to both the Vp and the spinal trigeminal nucleus (Vo, Vi and/or Vc) by way of axon collaterals.Yun-Qing Li is on leave from the Department of Anatomy, The Fourth Military Medical University, Xian, People's Republic of China     

Modulation of IA currents by capsaicin in rat trigeminal ganglion neurons

When capsaicin, the pungent compound in hot pepper, is applied to epithelia it produces pain, allodynia, and hyperalgesia. We investigated, using whole cell path clamp, whether some of these responses induced by capsaicin could be a consequence of capsaicin blocking I(A) currents, a reduction in which, such as occurs in injury, increases neuronal excitability. In capsaicin-sensitive (CS) rat trigeminal ganglion (TG) neurons, capsaicin inhibited I(A) currents in a dose-dependent manner. I(A) currents were reduced 49% by 1 microM capsaicin. In capsaicin-insensitive (CIS) rat TG neurons, or small-diameter mouse VR1-/- neurons, 1 microM capsaicin inhibited I(A) currents 9 and 3%, respectively. These data suggest that in CS neurons the vast majority of the capsaicin-induced inhibition of I(A) currents occurs as a consequence of the activation of vanilloid receptors. Capsaicin (1 microM) did not alter the I(A) conductance-voltage relationship but shifted the inactivation-voltage curve about 15 mV to hyperpolarizing voltages, thereby increasing the number of inactivated I(A) channels at the resting potential. I(A) currents were relatively unaffected by 1 mM CTP-cAMP or 500 nM phorbol-12, 13-dibuterate (a protein kinase C agonist) but were inhibited by 20-30% with either 1 mM CTP-cGMP or 25 microM N-(6-aminohexyl)-5-chloro-1-napthalenesulfonamide HCl (a calcium-calmodulin kinase inhibitor). In the presence of 0.5 microM KT5823, an inhibitor of protein kinase G (PKG) pathways, 1 microM capsaicin inhibited I(A) by only 26%. In summary, in CS neurons, capsaicin decreases I(A) currents through the activation of vanilloid receptors. That activation, partially through the activation of cGMP-PKG and calmodulin-dependent pathways should result in increased excitability of capsaicin-sensitive nociceptors.     

Effects of trigeminal ganglion stimulation on unit activity of ventral cochlear nucleus neurons

The trigeminal ganglion sends a projection to the granule and magnocellular regions of the ventral cochlear nucleus (VCN; [J Comp Neurol 419 (2000) 271]), as well as to the cochlea ([Neuroscience 79 (1997) 605; Neuroscience 84 (1998a) 559]). We investigated the effects of electrically stimulating the trigeminal ganglion on unit responses in the guinea-pig VCN. Responses consisted of one, two or more phases of excitation, sometimes followed by a longer inhibitory phase. The latencies to the first excitation peak ranged between 5 and 17 ms from the onset of stimulation. These responses were preceded by a slow wave potential evoked by the stimulation. Applying kainic acid, which eliminates VIIIth nerve responses, diminished the firing rates of VCN units to trigeminal stimulation, and increased their first spike latencies. Cochlear destruction had a similar effect. The responses in VCN evoked by trigeminal ganglion stimulation therefore appear to result from direct stimulation of the trigeminal ganglion-cochlear nucleus pathway, as well as modulation by the trigeminal ganglion-cochlear pathway. Alternatively, a reduction in spontaneous rate of VCN neurons by removal of VIIIth nerve input could explain the decreased response to trigeminal stimulation after cochlear manipulations. The modulation of firing rate in second order auditory neurons by first order somatosensory neurons could influence central auditory targets and may be involved in generating or modulating perceptions of phantom sounds which can be modified by manipulations of somatic regions of the head and neck ("somatic tinnitus").     

Consequences of 4-aminopyridine applications to trigeminal root ganglion neurons

1. The effects of 4-aminopyridine (4-AP) on the electrical properties of 30 trigeminal root ganglion (TRG) neurons were determined from the membrane voltage responses to step and sinusoidal current injections using intracellular microelectrode techniques in in vitro slice preparations (guinea pigs). 2. Comparisons of results from 4-AP applications (0.05-5 mM) with those from tetraethylammonium (TEA) applications (0.1-10 mM) revealed very different actions of these agents. Both agents produced an increase in input resistance and a decrease in threshold for spike generation. Applications of 4-AP increased subthreshold oscillations of the membrane potential and enhanced the repetitive spike firing evoked by intracellular injections of current pulses. However, TEA applications blocked the potential oscillations and did not exaggerate repetitive spike discharges. Spontaneous spike activity or bursts were observed in four neurons that received 4-AP applications. 3. Membrane properties were determined in 20 of the 30 neurons by fitting impedance data in the frequency domain with a four-parameter membrane model by the use of computer-intensive techniques. In the majority of neurons, the time-invariant and time-dependent membrane conductances decreased during 4-AP application. The time constant for the time-dependent conductance also decreased, suggesting that the closing of K+-channels was facilitated in the membrane. 4. Applications of 4-AP in a dose range of 50 microM-5 mM produced rapid (approximately tens of seconds) responses of the neurons, resulting in a dose-dependent increase of the impedance magnitude functions and in a leftward shift of the resonant "humps" to lower frequencies. This shift indicates that the TRG neuronal membrane is capable of producing large voltage responses to current inputs at low frequencies. Recovery from the effects of 4-AP was slow (usually greater than 30 min). 5. Applications of 4-AP at high doses (greater than or equal to 1 mM) and at various imposed membrane potentials in four neurons resulted in poorly reversible unspecific changes in certain membrane parameters (increased input capacitance and conductance) and an insensitivity of the input conductance to the imposed membrane potential. These effects could be interpreted as membrane breakdown. 6. The tendencies of TRG neurons to fire repetitively and in bursts of spikes during 4-AP application result from the increased oscillatory behavior of their membrane potentials and changes in membrane resonance induced by presumed blockade of K+ channels.(ABSTRACT TRUNCATED AT 400 WORDS)     

Responses of neurons in the spinal nucleus of the trigeminal nerve to electrical stimulation of the dura mater of the rat brain

The pathogenesis of migraine is based on the aseptic inflammation of dura mater tissues surrounding the large cranial vessels, such as the superior sagittal sinus. This inflammation develops in conditions of antidromic activation of sensory terminals of the trigeminal nerve and is accompanied by changes in the responses of neurons in the spinal nucleus of the trigeminal nerve to electrical stimulation of the superior sagittal sinus. However, the characteristics of the responses of these neurons to this stimulation have received virtually no study. Experiments on anesthetized rats were performed with recording of the responses of 387 neurons in the spinal nucleus of the trigeminal nerve to electrical stimulation of the superior sagittal sinus. The results showed that the responses of neurons to this stimulation was biphasic, consisting of a short initial phase with a latent period of 7–19 (11.4 ± 0.17) msec, followed by a longer-lived discharge with a latent period of 20–50 (34.2 ± 0.8) msec. It is suggested that the first phase reflects orthodromic activation of perivascular A and C fibers of the trigeminal nerve, while the second phase is associated with activation of meningeal C fibers with low conduction velocities and/or secondary activation of the perivascular sensory endings of the trigeminal nerve by algogenic and vasoactive substances released from them during antidromic activation. These changes seen in animal experiments may serve as an indicator of the efficacy of antimigraine agents.Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 90, No. 1, pp. 3–10, January, 2004.     

Substance P potentiates 5-HT3 receptor-mediated current in rat trigeminal ganglion neurons

The present study aimed to investigate the interaction between the coexistent SP receptor and 5-HT3 receptor in trigeminal ganglion (TG) neurons using whole-cell patch clamp technique. The majority of the neurons examined responded to 5-HT with an inward current (I5-HT) (78.2%, 79/101) that could be blocked by 5-HT3 receptor antagonist, ICS-205,930. The I5-HT was potentiated by preapplication of SP (10(-10) to 10(-8) M) in most 5-HT-sensitive cells(78.5%, 62/79). Coapplication of SP and GR-82334, antagonist of NK1 receptor, had no enhancing effect on I5-HT. The concentration-response curves for 5-HT with and without SP preapplication show that: (1) the threshold 5-HT concentrations with and without SP preapplication are basically the same, while SP preapplication increased the maximal value of I5-HT by 38.0% of its control; (2) the EC50 values of the curves with and without SP pretreatment are very close, i.e. 1.89 x 10(-5) M and 2.08 x 10(-5) M (P > 0.1; n = 9), respectively. Intracellular dialysis of GDP-beta-S, a non-hydrolyzable GDP analog, and GF-109203X, a selective protein kinase C inhibitor, removed the SP potentiation of I5-HT. These results may offer a clue to understanding the mechanism underlying the generation and/or regulation of peripheral pain caused by tissue damage inflammation, etc.     

Responses of nucleus reticularis tegmenti pontis neurons to vestibular stimulation in the rat

Summary Forty-nine neurons were recorded in the nucleus reticularis tegmenti pontis (NRTP) during horizontal vestibular and/or optokinetic stimulation in immobilized pigmented rats. During optokinetic stimulation, the response of NRTP neurons was either unidirectional (51%) or bidirectional (49%). Histological reconstruction showed that unidirectional neurons were located in the dorsal-medial part of NRTP, and bidirectional neurons in the lateral part. All neurons exhibited a response during pure vestibular sinusoidal stimulation in the frequency range 0.025 Hz-0.2 Hz. NRTP neurons were divided into two groups according to their threshold to vestibular stimulation. Group A neurons had a low threshold, a low spontaneous activity and their firing frequency slowly increased with acceleration. Group B neurons showed opposite characteristics. Phase and gain analysis suggested that NRTP neurons carry a head velocity signal. After hemiflocculectomy, the gain of the vestibular response of contralateral NRTP neurons increased. From these data, the role of NRTP in the horizontal vestibulo-oculomotor is discussed.Supported by DGRST 79.7.1012     

Responses of feline trigeminal spinal tract nucleus neurons to stimulation of the middle meningeal artery and sagittal sinus

1. Extracellular single-unit activity was recorded from 250 trigeminal (V) spinal tract nucleus neurons that were excited by electrical stimulation of the middle meningeal artery (MMA) and/or sagittal sinus (SS) in anesthetized cats. One hundred and thirty of these neurons were located in the V subnucleus caudalis (Vc), and the remaining 120 neurons were located in the V subnucleus oralis (Vo) or rostral part of the V subnucleus interpolaris (Vi). In many cases these neurons were also examined for the existence of orofacial receptive fields (RFs) by applying mechanical stimuli to the orofacial region. 2. The mean minimum latencies to suprathreshold electrical stimulation of the MMA and the SS were similar for Vc and Vo/Vi neurons. Excitation of Vc neurons occurred at latencies of 14.3 +/- 1.0 (n = 102) and 17.4 +/- 1.6 ms (n = 36) to MMA and SS stimulation, respectively. Excitation of Vo/Vi neurons occurred at latencies of 12.4 +/- 0.9 (n = 86) and 16.4 +/- 1.1 ms (n = 58) to MMA and SS stimulation, respectively. These latencies correspond to mean conduction velocities of approximately 5.2 and 4.0 m/s to MMA and SS stimulation, respectively. 3. Mechanical stimulation of the vessels evoked neuronal responses in five of eight MMA-activated neurons tested and three of five SS-activated neurons tested. 4. Almost all of the neurons tested (127 of 131) had peripheral RFs, and all were in the orofacial region. Nearly all (95%) Vc neurons had RFs within or including the ophthalmic facial region. The RFs of most (67%) Vo/Vi neurons also had RFs within or including ophthalmic regions, but in some cases were only within maxillary and/or mandibular regions. 5. Most of the Vc neurons (87%) were classified on the basis of their cutaneous inputs as nociceptive. The incidence of nociceptive neurons in Vo/Vi was also high (61%), although less than in Vc. In both the Vc and Vo/Vi neuronal populations, MMA-activated neurons were more likely to have nociceptive peripheral inputs than SS-activated neurons. 6. Histological reconstruction of recording sites indicated that the MMA- and/or SS-activated Vc neurons were concentrated in the lateral half of laminae III-V. The responsive neurons at the level of Vo/rostral Vi were not confined to any particular portion of these subnuclei. 7. These findings indicate that sensory afferents that innervate the dural arteries and venous sinuses are capable of activating neurons throughout the V spinal tract nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Responses of visual cortical neurons to nociceptive stimulation

Conclusion Nociceptive excitation, the basis for the formation of the animal's defensive emotional state, and arising in response to the action of nociceptive stimulation, activates cortical neurons through neurochemical mechanisms that differ from the mechanisms of transmission of excitation in sensory projection systems.Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti, Vol. 29, No. 2, pp. 389–395, March–April, 1979.     

人胚胎三叉神经节神经元的躯体定位

目的:应用1,1′-双十八烷-3,3,3′,3′-四甲基吲哚羰花青-高氯酸盐(DiI)追踪技术研究人胚胎发出眼神经、上颌神经及下颌神经的神经元在三叉神经节内的解剖定位.方法:对胎龄20周以上的6个胎儿标本进行灌注固定后,分别于标本的眼、上颌、下颌神经处植入DiI染色晶体.37℃恒温箱放置3个月,等待DiI染色晶体扩散,再根据神经走向切片,荧光显微镜下观察发出3支神经的神经元在三叉神经节内的解剖定位.结果:人胚胎三叉神经节是沿内外方向分布的细胞团.发出眼神经的神经元位于神经节的前内侧,下颌神经的神经元位于神经节的后外侧,上颌神经的神经元位于两者之间.结论:人胚胎发出眼神经、上颌神经及下颌神经的神经元在三叉神经节内以内外局部定位方式排列.     

Electrophysiological responses of trigeminal root ganglion neurons in vitro

The membrane electrical properties of neurons and their responses to endogenous compounds or other neuroactive substances were investigated in vitro with intracellular recording techniques in slices of trigeminal root ganglia of guinea-pigs. The mean resting membrane potential of these neurons was -60 mV. Intracellular injections of hyperpolarizing current pulses evoked time-dependent rectification with varying degrees of dependence on membrane voltage in 107 of 110 neurons. Membrane potential oscillations were observed following the termination of the hyperpolarizing pulses and after similar injections of depolarizing current. This phenomenon appeared to be voltage-dependent at levels that were subthreshold for spike genesis; the more pronounced oscillations were evident at the more depolarized levels and were insensitive to tetrodotoxin applications. Two groups of neurons could be distinguished on the basis of certain characteristics in their action potentials. The majority exhibited short duration (0.6 ms) spikes with mean amplitude of 72 mV in response to intracellular depolarizing current. The brief (3 ms) afterhyperpolarizations that followed such spikes were blocked by intracellular injections of Cs+ or by bath applications of tetraethylammonium. Action potentials in the minority group exhibited a hump in their repolarization phase. The humped spikes had a mean peak amplitude of 78 mV and a longer duration (2 ms). Both the duration (6 ms) and the amplitude (16 mV) of the afterhyperpolarization were significantly greater in this latter group of neurons. Some fast spikes were easily blocked whereas others, including humped spikes, were resistant to tetrodotoxin (10(-6) M). Spikes which were resistant, were also not affected by perfusion with Co2+ (10(-3) M) and were reduced in amplitude during perfusion with Na+-deficient solution. Bath applications of S-glutamate (10(-4)-10(-2) M) depolarized only two of ten neurons by less than 3 mV. Similarly, 5-hydroxytryptamine produced a small depolarization in only two of thirteen neurons. Perfusion of gamma-aminobutyrate (10(-5)-10(-2) M) resulted in an increase in input conductance that waned despite continued application and was associated with a depolarization (2-14 mV) in 44/50 neurons. In some neurons, gamma-aminobutyrate application enhanced their repetitive firing ability, possibly as a result of the increased oscillatory behavior of the membrane at certain depolarized potentials. The effects of gamma-aminobutyrate were blocked by the GABAA-receptor antagonist, bicuculline (10(-4) M) but were unaffected by the GABAB-receptor agonist, baclofen (10(-4) M).(ABSTRACT TRUNCATED AT 400 WORDS)     

Osmolality-induced tuning of action potentials in trigeminal ganglion neurons

The present study explored the effect of anisotonicity on action potential (AP) in cultured trigeminal ganglion (TG) neurons. We demonstrate that the number of evoked APs was increased by both hypo- and hypertonic treatment. Transient Receptor Potential Vanilloid 4 receptor (TRPV4) activator increased the number of APs, but only hypotonic-response was markedly blocked in TRPV4^−/− mice. Additionally, inhibition of PKC attenuated hypotonicity-induced increase, whereas antagonism of PKA attenuated hypertonicity-response. We conclude that anisotonicity increases excitability of nociceptors, which might be involved in anisotonicity-induced nociception. The increase of APs by hypo- and hypertonicity is mediated through different receptor and intracellular signaling pathways.