Principal Sensory Nucleus Research Articles

C-Fos induction in the brainstem following electrical stimulation of the trigeminal ganglion of chronically mandibular nerve-transected rats

Neuronal excitability in the trigeminal sensory nuclei (TSN) changes after nerve transection. We examined the effects of chronic transection of the trigeminal nerve on the c-Fos-immunoreactivity in the TSN induced 2 h after 10 min of electrical stimulation of the trigeminal ganglion (TG) at C-fiber activating condition (1.0 mA, 5 ms, 5 Hz) in urethane-anesthetized rats. In the non-transected control rats, stimulation of the TG induced c-Fos-immunoreactive cells (c-Fos-IR cells) mostly in superficial layers (VcI/II) of the nucleus caudalis (Vc) in its full extent along the dorsomedial–ventrolateral axis, but modestly in the rostral TSN above the obex, the principal, oral, and interpolar nuclei. Three days, 1, 2, or 3 weeks after transection of the inferior alveolar (IAN), infraorbital, or masseteric nerves, the stimulation of the TG induced c-Fos-IR cells in the central terminal fields of the transected nerve in the rostral TSN and magnocellular zone of the Vc. However, the number of c-Fos-IR cells in the VcI/II decreased inside the central terminal fields of the transected nerve and increased outside the fields. These results indicate that transection of the trigeminal nerve increases the excitability of TSN neurons that receive inputs from injured mechanoreceptors and uninjured nociceptors, but decreases it from injured nociceptors. The altered c-Fos responses may imply mechanisms of neuropathic pain seen after nerve injury.

Assessment of the wind-up phenomenon in the equine nociceptive trigeminal system

Repeated sub-threshold nociceptive electrical stimulation resulting in temporal summation of the limb nociceptive withdrawal reflex is a well-established non-invasive model to investigate the wind-up phenomenon in horses. Due to structural similarities of the trigeminal sensory nucleus to the dorsal horn of the spinal cord, temporal summation should be evoked by repeated transcutaneous electrical stimulation of trigeminal afferents. To evaluate this hypothesis repeated transcutaneous electrical stimulation was applied to the supraorbital and infraorbital nerves of 10 horses. Stimulation intensities varied between 0.5 and 1.3 times the trigemino-cervical reflex threshold defined for single stimulation. Evoked electromyographic activity of the orbicularis oculi, splenius and cleidomastoideus muscles was recorded and the signals analysed in the previously established epochs typical to the early and late component of the blink reflex and to the trigemino-cervical reflex. Behavioural reactions were evaluated with the aid of numerical rating scale.The nociceptive late component and the trigemino-cervical reflex were not elicited by sub-threshold intensity repeated transcutaneous electrical stimulation. Furthermore, the median reflex amplitude for the 10 horses showed a tendency to decline over the stimulation train so temporal summation of afferent trigeminal inputs could not be observed. Therefore, the modulation of trigeminal nociceptive processing attributable to repeated Aδ fibre stimulations seems to differ from spinal processing of similar inputs as it seems to have an inhibitory rather than facilitatory effect. Further evaluation is necessary to highlight the underlying mechanism.

Differential expression of VGLUT1 or VGLUT2 in the trigeminothalamic or trigeminocerebellar projection neurons in the rat

The vesicular glutamate transporters, VGLUT1 and VGLUT2, reportedly display complementary distribution in the rat brain. However, co-expression of them in single neurons has been reported in some brain areas. We previously found co-expression of VGLUT1 and VGLUT2 mRNAs in a number of single neurons in the principal sensory trigeminal nucleus (Vp) of the adult rat; the majority of these neurons sent their axons to the thalamic regions around the posteromedial ventral nucleus (VPM) and the posterior nuclei (Po). It is well known that trigeminothalamic (T-T) projection fibers arise not only from the Vp but also from the spinal trigeminal nucleus (Vsp), and that trigeminocerebellar (T-C) projection fibers take their origins from both of the Vp and Vsp. Thus, in the present study, we examined the expression of VGLUT1 and VGLUT2 in Vp and Vsp neurons that sent their axons to the VPM/Po regions or the cortical regions of the cerebellum. For this purpose, we combined fluorescence in situ hybridization (FISH) histochemistry with retrograde tract-tracing; immunofluorescence histochemistry was also combined with anterograde tract-tracing. The results indicate that glutamatergic Vsp neurons sending their axons to the cerebellar cortical regions mainly express VGLUT1, whereas glutamatergic Vsp neurons sending their axons to the thalamic regions express VGLUT2. The present data, in combination with those of our previous study, indicate that glutamatergic Vp neurons projecting to the cerebellar cortical regions express mainly VGLUT1, whereas the majority of glutamatergic Vp neurons projecting to the thalamus co-express VGLUT1 and VGLUT2.

Morphology and connections of intratrigeminal cells and axons in the macaque monkey

Trigeminal primary afferent fibers have small receptive fields and discrete submodalities, but second order trigeminal neurons often display larger receptive fields with complex, multimodal responses. Moreover, while most large caliber afferents terminate exclusively in the principal trigeminal nucleus, and pars caudalis (sVc) of the spinal trigeminal nucleus receives almost exclusively small caliber afferents, the characteristics of second order neurons do not always reflect this dichotomy. These surprising characteristics may be due to a network of intratrigeminal connections modifying primary afferent contributions. This study characterizes the distribution and morphology of intratrigeminal cells and axons in a macaque monkeys. Tracer injections centered in the principal nucleus (pV) and adjacent pars oralis retrogradely labeled neurons bilaterally in pars interpolaris (sVi), but only ipsilaterally, in sVc. Labeled axons terminated contralaterally within sVi and caudalis. Features of the intratrigeminal cells in ipsilateral sVc suggest that both nociceptive and non-nociceptive neurons project to principalis. A commissural projection to contralateral principalis was also revealed. Injections into sVc labeled cells and terminals in pV and pars oralis on both sides, indicating the presence of bilateral reciprocal connections. Labeled terminals and cells were also present bilaterally in sVi and in contralateral sVc. Interpolaris injections produced labeling patterns similar to those of sVc. Thus, the rostral and caudal poles of the macaque trigeminal complex are richly interconnected by ipsilateral ascending and descending connections providing an anatomical substrate for complex analysis of oro-facial stimuli. Sparser reciprocal crossed intratrigeminal connections may be important for conjugate reflex movements, such as the corneal blink reflex.

Complementary processing of haptic information by slowly and rapidly adapting neurons in the trigeminothalamic pathway. Electrophysiology, mathematical modeling and simulations of vibrissae-related neurons

Tonic (slowly adapting) and phasic (rapidly adapting) primary afferents convey complementary aspects of haptic information to the central nervous system: object location and texture the former, shape the latter. Tonic and phasic neural responses are also recorded in all relay stations of the somatosensory pathway, yet it is unknown their role in both, information processing and information transmission to the cortex: we don't know if tonic and phasic neurons process complementary aspects of haptic information and/or if these two types constitute two separate channels that convey complementary aspects of tactile information to the cortex. Here we propose to elucidate these two questions in the fast trigeminal pathway of the rat (PrV-VPM: principal trigeminal nucleus-ventroposteromedial thalamic nucleus). We analyze early and global behavior, latencies and stability of the responses of individual cells in PrV and medial lemniscus under 1–40 Hz stimulation of the whiskers in control and decorticated animals and we use stochastic spiking models and extensive simulations. Our results strongly suggest that in the first relay station of the somatosensory system (PrV): (1) tonic and phasic neurons process complementary aspects of whisker-related tactile information (2) tonic and phasic responses are not originated from two different types of neurons (3) the two responses are generated by the differential action of the somatosensory cortex on a unique type of PrV cell (4) tonic and phasic neurons do not belong to two different channels for the transmission of tactile information to the thalamus (5) trigeminothalamic transmission is exclusively performed by tonically firing neurons and (6) all aspects of haptic information are coded into low-pass, band-pass, and high-pass filtering profiles of tonically firing neurons. Our results are important for both, basic research on neural circuits and information processing, and development of sensory neuroprostheses.

Pattern of tau hyperphosphorylation and neurotransmitter markers in the brainstem of senescent tau filament forming transgenic mice

The early occurrence of brainstem-related symptoms, e.g. gait and balance impairment, apathy and depression in Alzheimer’s disease patients suggests brainstem involvement in the initial pathogenesis. To address the question whether tau filament forming mice expressing mutated human tau mirror histopathological changes observed in Alzheimer brainstem, the degree and distribution of neurofibrillary lesions as well as the pattern of cholinergic and monoaminergic neurons were investigated. The expression of the human tau transgene was observed in multiple brainstem nuclei, particularly in the magnocellular reticular formation, vestibular nuclei, cranial nerve motor nuclei, sensory trigeminal nerve nuclei, inferior and superior colliculi, periaqueductal and pontine gray matter, and the red nucleus. Most of the human tau-immunoreactive cell groups also showed tau hyperphosphorylation at the epitopes Thr231/Ser235 and Ser202/Thr205, while abnormal tau phosphorylation at the epitope Ser422 or silver stained structures were almost totally lacking. We found no obvious differences in distribution and density of cholinergic and monoaminergic neurons between tau-transgenic and wild type mice. Although numerous brainstem nuclei in our model expressed human tau protein, the development of neurofibrillary tangles, neuropil threads and ghost tangles was rare and likewise its distribution differed largely from Alzheimer′s disease pattern. The number of monoaminergic neurons remained unchanged in the transgenic mice, while monoaminergic nuclei in Alzheimer brainstem showed a distinct neuronal loss. However, the distribution of pretangle-affected neurons in the tau-transgenic mice partly resembled those seen in progressive supranuclear palsy, presenting these animals as a model to examine brainstem pathogenesis of progressive supranuclear palsy.

Cooperative slit and netrin signaling in contralateralization of the mouse trigeminothalamic pathway

Ascending somatosensory pathways are crossed pathways representing each side of the body in the contralateral neocortex. The principal sensory nucleus of the trigeminal nerve (PrV) relays the facial sensations to the contralateral somatosensory cortex via the ventrobasal thalamus. In the companion article (Kivrak and Erzurumlu [2012] J. Comp. Neurol. 12-0013) we described the normal development of the trigeminal lemniscal pathway in the mouse. In this study we investigated the role of midline axon navigation signals, the netrin and slit proteins. In situ hybridization assays revealed that both netrin and slit mRNAs are expressed along the midline facing the PrV axons and their receptors are expressed in developing PrV neurons. In wild-type mouse embryos, PrV axons cross the midline and take a sharp rostral turn heading toward the contralateral thalamus. Examination of trigeminal lemniscal axons in dcc knockout mice revealed absence of midline crossing between E11 and E15. However, a few axons crossed the midline at E17 and reached the contralateral thalamus, resulting in a bilateral PrV lemniscal pathway at P0. We also found that slit1, -2 or -3 single or double knockout mice have impaired development of the trigeminal-lemniscal pathway. These include axon stalling along the midline, running within the midline, and recrossing of axons back to the site of origin. Collectively, our studies indicate a cooperative role for netrin and slit proteins in midline attraction and crossing behavior of the ascending facial somatosensory projections during development.

Of apples and oranges: GABA and glutamate transmission in neurones of the nucleus tractus solitarii could not be more different

The nucleus tractus solitarii (NTS) is the principal visceral sensory nucleus in the brain and comprises neurochemically and biophysically distinct neurons located in the dorsomedial medulla oblongata. The NTS conveys information from the gustatory, cardiorespiratory, oesophageal and subdiaphragmatic gastrointestinal visceral systems that is subsequently assimilated with homeostatic signals arriving from an impressive network of connections from other integrative centres of the pons, diencephalon and forebrain. Myelinated A-type and unmyelinated C-type sensory afferent fibres from the trigeminal, facial, glossopharyngeal and vagus nerve form the tractus solitarius (TS) and their terminals overlap along the rostro-caudal extent of the NTS. Regardless of their origin or type, though, afferent fibres entering the brainstem activate NTS neurones via the release of excitatory neurotransmitters, mainly glutamate (Andresen & Kunze, 1994; Baptista et al. 2005). Perhaps the most prominent neurochemical phenotypes encountered in the NTS are GABAergic and glutamatergic, which provide postsynaptic inhibitory and excitatory influences to adjacent motor nuclei such as the dorsal motor nucleus of the vagus, the nucleus ambiguus and the ventrolateral medulla. An impressive amount of work has been conducted in recent times by Andresen's group to characterize the glutamatergic transmission from the tractus solitarius onto NTS neurones. This group has used sophisticated electrophysiological analyses to demonstrate that glutamatergic excitatory transmission is highly effective, has an almost unfailing unitary transmission from primary sensory fibres, and can be associated with forms of synaptic plasticity that are unique to autonomic regulation (Peters et al. 2010). Earlier work by Mifflin (Mifflin, 1996; Mifflin & Felder, 1988) and Smith (Smith et al. 1998), however, showed that TS stimulation can evoke a sequence of excitatory responses that were followed by inhibitory currents, indicating the possible presence of a local GABAergic interneurone. This inhibitory interneurone, however, has not yet been demonstrated with anatomical tools. (Note that the presence of GABAergic neurons within the NTS is not in doubt. What remains to be confirmed, however, is the anatomical proof of their interneurone status, i.e. a neurone that does not project beyond the NTS itself.) The publication by McDougall and Andresen (2012) in this issue of The Journal of Physiology utilises an isolated horizontal brainstem slice in which TS fibres can be stimulated to evoke unitary currents in second order NTS neurones; these currents can then be identified as either mono- or polysynaptic. The authors showed that, in contrast to glutamatergic currents, monosynaptic GABAergic currents are evoked by focal stimulation disproportionately rarely and only following a stimulus that was located in close proximity to the recorded neurone. In addition to the high failure rate, large amplitude variability, low calcium sensitivity and modest frequency-dependent depression, the monosynaptic nature of these GABAergic currents was confirmed by a low jitter (albeit greater than that of glutamatergic synapses) and lack of sensitivity to glutamatergic receptor blockade. Overall, these data suggest rather weak GABAergic transmission whose efficacy must rely on converging activation of inhibitory fibres. These characteristics suggest strongly that the GABA release properties are fundamentally different from the powerful and synaptically reliable glutamatergic inputs impinging onto NTS neurones. When taken together with previous observations reporting that each TS-driven glutamatergic input targets individual NTS neurons selectively, and that the physiological and pharmacological responses of NTS neurones are target-coded (Bailey et al. 2006; Browning et al. 2011; Jin et al. 2004), the overall picture of the NTS strengthens the image of a hyperorganized nucleus with ‘task matching’ capabilities conferring an ability to cope with widely variable demands, both in terms of timing as well as duration and modulation of a response. Furthermore, this type of cellular organization within the NTS should force us, once again, to be extremely cautious in the often simplistic physiological interpretation of observations, often driven by high intensity stimuli, that analyse a single outcome such as cFos expression or calcium oscillations, which provide limited information regarding the excitability of neurones and provide no information at all regarding inhibitory transmission. Several questions remain to be answered before one can elucidate the role of these synaptic connections. For example, what are the inputs that activate these second order GABAergic neurones? What is their origin? Are they anatomically defined interneurones, or do they project beyond the NTS? Are they related to a particular pathway? Do they have a specific physiological role or are they ‘simply’ a low pass filter for excitatory TS-driven inputs?

Response properties of temporomandibular joint mechanosensitive neurons in the trigeminal sensory complex of the rabbit

The neurophysiological properties of neurons sensitive to TMJ movement (TMJ neurons) in the trigeminal sensory complex (Vcomp) during passive movement of the isolated condyle were examined in 46 rabbits. Discharges of TMJ neurons from the rostral part of the Vcomp were recorded with a microelectrode when the isolated condyle was moved manually and with a computer-regulated mechanostimulator. A total of 443 neurons responding to mechanical stimulation of the face and oral cavity were recorded from the brainstem. Twenty-one TMJ neurons were detected rostrocaudally from the dorsal part of the trigeminal principal sensory nucleus (NVsnpr), subnucleus oralis of the trigeminal spinal nucleus, and reticular formation surrounding the trigeminal motor nucleus. Most of the TMJ neurons were located in the dorso-rostral part of the NVsnpr. Of the TMJ units recorded, 90 % were slowly adapting and 26 % had an accompanying resting discharge. The majority (86 %) of the TMJ units responded to the movement of the isolated condyle in the anterior and/or ventral directions, and half were sensitive to the condyle movement in a single direction. The discharge frequencies of TMJ units increased as the condyle displacement and constant velocity (5 mm/s) increased within a 5-mm anterior displacement of the isolated condyle. Based on these results, we conclude that sensory information is processed by TMJ neurons encoding at least joint position and displacement in the physiological range of mandibular displacement.

Distinct regional and subcellular localization of the actin‐binding protein filamin a in the mature rat brain

Filamin A (FLNa) is an actin-binding protein that regulates cell motility, adhesion, and elasticity by cross-linking filamentous actin. Additional roles of FLNa include regulation of protein trafficking and surface expression. Although the functions of FLNa during brain development are well studied, little is known on its expression, distribution, and function in the adult brain. Here we characterize in detail the neuroanatomical distribution and subcellular localization of FLNa in the mature rat brain, by using two antisera directed against epitopes at either the N' or the C' terminus of the protein, further validated by mRNA expression. FLNa was widely and selectively expressed throughout the brain, and the intensity of immunoreactivity was region dependent. The most intensely FLNa-labeled neurons were found in discrete neuronal systems, including basal forebrain structures, anterior nuclear group of thalamus, and hypothalamic parvocellular neurons. Pyramidal neurons in neocortex and hippocampus and magnocellular cells in basolateral amygdaloid nucleus were also intensely FLNa immunoreactive, and strong FLNa labeling was evident in the pontine and medullary raphe nuclei and in sensory and spinal trigeminal nuclei. The subcellular localization of FLNa was evaluated in situ as well as in primary hippocampal neurons. Punctate expression was found in somata and along the dendritic shaft, but FLNa was not detected in dendritic spines. These subcellular distribution patterns were recapitulated in hippocampal and neocortical pyramidal neurons in vivo. The characterization of the expression and subcellular localization of FLNa may provide new clues to the functional roles of this cytoskeletal protein in the adult brain.

Convergent Pre-motoneuronal Inputs to Single Trigeminal Motoneurons

Because pre-motor neurons targeting trigeminal motoneurons are located in various regions, including the supratrigeminal (SupV) and intertrigeminal (IntV) regions, the principal sensory trigeminal nucleus (PrV), and the region dorsal to the PrV (dRt), a single trigeminal motoneuron may receive differential convergent inputs from these regions. We thus examined the properties of synaptic inputs from these regions to masseter motoneurons (MMNs) and digastric motoneurons (DMNs) in brainstem slice preparations obtained from P1-5 neonatal rats, using whole-cell recordings and laser photolysis of caged glutamate. Photostimulation of multiple regions within the SupV, IntV, PrV, and dRt induced post-synaptic currents (PSCs) in 14 of 19 MMNs and 18 of 26 DMNs. Furthermore, the stimulation of the lateral SupV significantly induced burst PSCs in MMNs more often than low-frequency PSCs in MMNs or burst PSCs in DMNs. Similar results were obtained in the presence of the GABAA receptor antagonist SR95531 and the glycine receptor antagonist strychnine. These results suggest that both neonatal MMNs and DMNs receive convergent glutamatergic inputs from the SupV, IntV, PrV, and dRt, and that the lateral SupV sends burst inputs predominantly to the MMNs. Such convergent pre-motoneuronal inputs to trigeminal motoneurons may contribute to the proper execution of neonatal oro-motor functions.

Somatotopic direct projections from orofacial areas of secondary somatosensory cortex to trigeminal sensory nuclear complex in rats

Little is known about the projections from the orofacial areas of the secondary somatosensory cortex (S2) to the pons and medulla including the second-order somatosensory neuron pools. To address this in rats, we first examined the distribution of S2 neurons projecting to the trigeminal principal nucleus (Vp) or oral subnucleus (Vo) of the trigeminal sensory nuclear complex (TSNC) after injections of a retrograde tracer, Fluorogold (FG), into five regions in the Vp/Vo which were responsive to stimulation of trigeminal nerves innervating the orofacial tissues. A large number of FG-labeled neurons were found with a somatotopic arrangement in the dorsal areas of S2 (orofacial S2 area). This somatotopic arrangement in the orofacial S2 area was shown to closely match that of the orofacial afferent inputs by recording cortical surface potentials evoked by stimulation of the trigeminal nerves. We then examined the morphology of descending projections from these electrophysiologically defined areas of the orofacial S2 to the pons and medulla after injections of an anterograde tracer, biotinylated dextranamine (BDA), into the areas. A large number of BDA-labeled axon fibers and terminals were seen only in some of the second-order somatosensory neuron pools, most notably in the contralateral TSNC, although the labeled terminals were not seen in certain rostrocaudal levels of the contralateral TSNC including the rostrocaudal middle level of the trigeminal interpolar subnucleus. The projections to the TSNC showed somatotopic arrangements in dorsoventral, superficial-deep and rostrocaudal directions. The somatotopic arrangements in the Vp/Vo closely matched those of the electrophysiologically defined central projection sites of the orofacial trigeminal afferents in the TSNC. The present results suggest that the orofacial S2 projects selectively to certain rostrocaudal levels of the contralateral TSNC, and the projections may allow the orofacial S2 to accurately modulate orofacial somatosensory transmission to higher brain centers including the orofacial S2 itself.

Distinct Development of the Trigeminal Sensory Nuclei in Platypus and Echidna

Both lineages of the modern monotremes have been reported to be capable of electroreception using the trigeminal pathways and it has been argued that electroreception arose in an aquatic platypus-like ancestor of both modern monotreme groups. On the other hand, the trigeminal sensory nuclear complex of the platypus is highly modified for processing tactile and electrosensory information from the bill, whereas the trigeminal sensory nuclear complex of the short-beaked echidna (Tachyglossus aculeatus) is not particularly specialized. If the common ancestor for both platypus and echidna were an electroreceptively and trigeminally specialized aquatic feeder, one would expect the early stages of development of the trigeminal sensory nuclei in both species to show evidence of structural specialization from the outset. To determine whether this is the case, we examined the development of the trigeminal sensory nuclei in the platypus and short-beaked echidna using the Hill and Hubrecht embryological collections. We found that the highly specialized features of the platypus trigeminal sensory nuclei (i.e. the large size of the principal nucleus and oral part of the spinal trigeminal nuclear complex, and the presence of a dorsolateral parvicellular segment in the principal nucleus) appear around the time of hatching in the platypus, but are never seen at any stage in the echidna. Our findings support the proposition that the modern echidna and platypus are derived from a common ancestor with only minimal trigeminal specialization and that the peculiar anatomy of the trigeminal sensory nuclei in the modern platypus emerged in the ornithorhynchids after divergence from the tachyglossids.

Unilateral Headache with Visual Aura from a Wallenberg Syndrome

Symptomatic migraine or migraine‐like headache due to a brainstem lesion is rarely reported, and limited reports of brainstem lesions with migraine‐like headache onset are all directed to the pons or midbrain [1, 2]. We describe a new onset of recurrent unilateral headache whose clinical features were similar to that of migraine with aura in a patient who had a lateral medullary infarction, i.e., Wallenberg syndrome.

Cluster-tic syndrome as the initial manifestation of multiple sclerosis

We report the case of a patient diagnosed as having cluster-tic syndrome as the initial manifestation of multiple sclerosis (MS). The patient’s headache bouts improved after treatment with antiepileptic drugs, steroids, and beta-interferon. Magnetic resonance imaging (MRI) scans showed a pontine demyelinating lesion involving the area of the trigeminal root inlet and main sensory nucleus. Neurophysiological studies correlated well with MRI lesions. The association between cluster-tic syndrome and MS is an exception, and the mechanism of the pain is still unknown; therefore, this case might suggest a pathophysiological relationship between the trigeminal main sensory nucleus and cluster-tic syndrome.

Physiological and anatomical evidence for an inhibitory trigemino-oculomotor pathway in the cat.

During blink down-phase, the levator palpebrae superioris (levator) muscle is inactivated, allowing the orbicularis oculi muscle to act. For trigeminal reflex blinks, the excitatory connections from trigeminal sensory nuclei to the facial nucleus have been described, but the pathway whereby the levator is turned off have not. We examined this question by use of both physiological and anatomical approaches in the cat. Intracellular records from antidromically activated levator motoneurons revealed that periorbital electrical stimulation produced bilateral, long latency inhibitory postsynaptic potentials (IPSPs). Central electrical stimulation of the principal trigeminal nucleus produced shorter latency IPSPs. Intracellular staining revealed that these motoneurons reside in the caudal central subdivision and have 10 or more poorly branched dendrites, which extend bilaterally into the surrounding supraoculomotor area. Axons penetrated in this region could be activated from periorbital and central electrodes. Neurons labeled from tracer injections into the caudal oculomotor complex were distributed in a crescent-shaped band that lined the ventral and rostral aspects of the pontine trigeminal sensory nucleus. Double-label immunohistochemical procedures demonstrated that these cells were not tyrosine hydroxylase-positive cells in the Kölliker-Fuse area. Instead, supraorbital nerve afferents displayed a similar crescent-shaped distribution, suggesting they drive these trigemino-oculomotor neurons. Anterograde labeling of the trigemino-oculomotor projection indicates that it terminates bilaterally, in and above the caudal central subdivision. These results characterize a trigemino-oculomotor pathway that inhibits levator palpebrae motoneurons in response to blink-producing periorbital stimuli. The bilateral distributions of trigemino-oculomotor afferents, levator motoneurons, and their dendrites supply a morphological basis for conjugate lid movements.

The role of a trigeminal sensory nucleus in the initiation of locomotion

While we understand how stimuli evoke sudden, ballistic escape responses, like fish fast-starts, a precise pathway from sensory stimulation to the initiation of rhythmic locomotion has not been defined for any vertebrate. We have now asked how head skin stimuli evoke swimming in hatchling frog tadpoles. Whole-cell recordings and dye filling revealed a nucleus of ∼20 trigeminal interneurons (tINs) in the hindbrain, at the level of the auditory nerve, with long, ipsilateral, descending axons. Stimulation of touch-sensitive trigeminal afferents with receptive fields anywhere on the head evoked large, monosynaptic EPSPs (∼5-20 mV) in tINs, at mixed AMPAR/NMDAR synapses. Following stimuli sufficient to elicit swimming, tINs fired up to six spikes, starting 4-8 ms after the stimulus. Paired whole-cell recordings showed that tINs produce small (∼2-6 mV), monosynaptic, glutamatergic EPSPs in the hindbrain reticulospinal neurons (descending interneurons, dINs) that drive swimming. Modelling suggested that summation of EPSPs from 18-24 tINs can make 20-50% of dINs fire. We conclude that: brief activity in a few sensory afferents is amplified by recruitment of many tINs; these relay summating excitation to hindbrain reticulospinal dINs; dIN firing then initiates activity for swimming on the stimulated side. During fictive swimming, tINs are depolarised and receive rhythmic inhibition but do not fire. Our recordings demonstrate a neuron-by-neuron pathway from head skin afferents to the reticulospinal neurons and motoneurons that drive locomotion in a vertebrate. This direct pathway, which has an important amplifier function, implies a simple origin for the complex routes to initiate locomotion in higher vertebrates.

Inducible Prrxl1‐CreERT2 recombination activity in the somatosensory afferent pathway

Prrxl1-CreER(T2) transgenic mice expressing tamoxifen-inducible Cre recombinase were generated by modifying a Prrxl1-containing BAC clone. Cre recombination activity was examined in Prrxl1-CreER(T2); Rosa26 reporter mice at various embryonic and postnatal stages. Pregnant mice were treated with a single dose of tamoxifen at embryonic day (E) 9.5 or E12.5, and X-gal staining was performed 2 days later. Strong X-gal staining was observed in the somatosensory ganglia (e.g., dorsal root and trigeminal ganglia) and the first central sites for processing somatosensory information (e.g., spinal dorsal horn and trigeminal nerve-associated nuclei). When tamoxifen was administered at postnatal day (P) 20 or in adulthood (P120), strong Cre recombination activity was present in the primary somatosensory ganglia, while weak Cre recombination activity was found in the spinal dorsal horn, mesencephalic trigeminal nucleus, principal sensory trigeminal nucleus, and spinal trigeminal nucleus. This mouse line provides a useful tool for exploring genes' functions in the somatosensory system in a time-controlled way.

The transcription factor, Lmx1b, promotes a neuronal glutamate phenotype and suppresses a GABA one in the embryonic trigeminal brainstem complex

Achieving an appropriate balance between inhibitory and excitatory neuronal fate is critical for development of effective synaptic transmission. However, the molecular mechanisms dictating such phenotypic outcomes are not well understood, especially in the whisker‐to‐barrel cortex neuraxis, an oft‐used model system for revealing developmental mechanisms. In trigeminal nucleus principalis (PrV), the brainstem link in the whisker–barrel pathway, the transcription factor Lmx1b marks glutamatergic cells. In PrV of Lmx1b knockout mice (−/−), initial specification of glutamatergic vs. GABAergic cell fate is normal until embryonic day 14.5. Subsequently, until the day of birth, glutamatergic markers (e.g., VGLUT2) stain significantly fewer PrV neurons, whereas, GABAergic markers (Pax2 and Gad1) stain significantly more PrV cells, notably in Lmx1b null PrV cells. These changes also occurred in Lmx1b/Bax double−/− mice, where PrV cells are rescued from Lmx1b−/− induced apoptosis; thus, effects upon excitatory/inhibitory cell ratios do not reflect a cell death confound. Electroporation‐induced ectopic expression of Lmx1b in an array of sites decreases numbers of neurons that express GABAergic markers, but increases VGLUT2+ cell numbers or stain intensity. Thus, Lmx1b is not involved in the initial specification of glutamatergic cell fate, but is essential for maintaining a glutamatergic phenotype. Other experiments suggest that Lmx1b acts to suppress Pax2, a promoter of GABAergic cell fate, in a cell‐autonomous manner, which may be a mechanism for maintaining a functional balance of glutamatergic and GABAergic cell types in development.

Spatial localization and projection densities of brainstem mossy fibre afferents to the forelimb C1 zone of the rat cerebellum.

The present study uses a double retrograde tracer technique in rats to examine the spatial localization and pattern of axonal branching in mossy fibres arising from three major sources in the medulla-the external cuneate nucleus, the sensory trigeminal nucleus and the reticular formation, to two electrophysiologically-identified parts of the cerebellar cortex that are linked by common climbing fibre input - the forelimb-receiving parts of the C1 zone in lobulus simplex and the paramedian lobule. In each experiment a small injection of rhodamine-tagged beads was injected into one cortical region and an injection of fluorescein-tagged beads was injected into the other region. The main findings were: (i) the proportion of double-labelled cells in each of the three precerebeller sources of mossy fibres was positively correlated with those in the inferior olive; and (ii) the C1 zone in lobulus simplex was found to receive a greater density of projections from all three sources of mossy fibres than the C1 zone in the paramedian lobule. These data suggest that two rostrocaudally separated but somatotopically corresponding parts of the C1 zone receive common mossy fibre and climbing fibre inputs. However, the differences in projection densities also suggest that the two parts of the zone differ in the extent to which they receive mossy fibre signals arising from the same precerebellar nuclei. This implies differences in function between somatotopically corresponding parts of the same cortical zone, and could enable a higher degree of parallel processing and integration of information within them.