The mesencephalic-hypoglossal nuclei loop as a possible central pattern generator for rhythmical whisking in rats.

It has been previously demonstrated that the Me5 nucleus is involved in the genesis of reflex activities at whisker pad level. Specific Me5 neurons, which provide sensory innervation of the macrovibrissae, are monosynaptically connected with small hypoglossal neurons innervating the extrinsic muscles that control macrovibrissal movements. Artificial whisking, induced by the electrical stimulation of the peripheral stump of the facial nerve and the electrical stimulation of the XII nucleus or the infraorbital nerve, induced evoked responses in the whisker pad extrinsic motor units, along with a significant increase in the electromyographic activity of the extrinsic pad muscles (Mameli et al. in Acta Oto-Laryngol 126:1334-1338, 2006; in Pfugers Arch Eur J Physiol 456:1189-1198, 2008; in Brain Res 1283:34-40, 2009; in Exp Brain Res 234:753-761, 2016). In anaesthetized rats, we evaluated the possible involvement of this Me5-XII loop in the genesis of rhythmical whisking. The anatomical findings showed that in addition to the ipsilateral, even the contralateral Me5 nucleus could be retrogradely labeled by the Dil tracer injected into the whisker pad of one side, they, furthermore, showed labeled axons extending across the midline between the two nuclei. The electrophysiological findings agreed with the neuroanatomical results, since the mechanical or artificially induced deflection of the whiskers of one side, evoked in the Me5 contralateral nucleus different patterns of responses. The hypothesis that the Me5-XII loops, along with their cross-linked relationship, could act as a "central generator" responsible for the stereotyped symmetrical pattern of macrovibrissal movements such as rhythmical whisking has been discussed.

Evidence for a trigeminal mesencephalic-hypoglossal nuclei loop involved in controlling vibrissae movements in the rat.

Previous studies performed in rats showed that the whisker-pad motor innervation involves not only the facial nerve, but also some hypoglossal neurons whose axons travel within the trigeminal infraorbital nerve (ION) and target the extrinsic muscles surrounding the whisker-pad macrovibrissae. Furthermore, the electrical stimulation of the ION induced an increase in the EMG activity of these muscles, while the hypoglossal nucleus stimulation elicited evoked potentials and single motor unit responses. However, the existence of a neural network able to involve the XIIth nucleus in macrovibrissae whisking control was totally unknown until now. Since other recent experiments demonstrated that: (1) the mesencephalic trigeminal nucleus (Me5) neurons respond to both spontaneous and artificial movements of macrovibrissae, and (2) the Me5 peripheral terminals provide a monosynaptic sensory innervation to the macrovibrissae, the present study was aimed at analyzing a possible role of the Me5 nucleus as a relay station in the sensory-motor loop that involves the XIIth nucleus neurons in rhythmic whisking control. Two tracers were used in the same animal: Fluoro Gold, which was injected into the whisker pad to retrogradely label the hypoglossal whisker-pad projection neurons, and Dil, which was instead injected into the Me5 to label its projections to these hypoglossal neurons. Results demonstrated that terminals of the Me5 neurons monosynaptically target the hypoglossal whisker-pad projection neurons. The functional role of this sensory-motor connection is discussed, with particular regard to a hypothesized proprioceptive reflex in whisker-pad extrinsic muscles that can be elicited by the activation of the Me5 macrovibrissae receptors.

Role of the trigeminal mesencephalic nucleus in rat whisker pad proprioception.

BACKGROUND: Trigeminal proprioception related to rodent macrovibrissae movements is believed to involve skin receptors on the whisker pad because pad muscles operate without muscle spindles. This study was aimed to investigate in rats whether the trigeminal mesencephalic nucleus (TMnu), which provides proprioceptive feedback for chewing muscles, may be also involved in whisker pad proprioception. METHODS: Two retrograde tracers, Dil and True Blue Chloride, were injected into the mystacial pad and the masseter muscle on the same side of deeply anesthetized rats to label the respective projecting sensory neurons. This double-labeling technique was used to assess the co-innervation of both structures by the trigeminal mesencephalic nucleus (TMnu).In a separate group of anesthetized animals, the spontaneous electrical activities of TMnu neurons were analyzed by extracellular recordings during spontaneous movements of the macrovibrissae. Mesencephalic neurons (TMne) were previously identified by their responses to masseter muscle stretching. Changes in TMne spontaneous electrical activities, analyzed under baseline conditions and during whisking movements, were statistically evaluated using Student's t-test for paired observations. RESULTS: Neuroanatomical experiments revealed different subpopulations of trigeminal mesencephalic neurons: i) those innervating the neuromuscular spindles of the masseter muscle, ii) those innervating the mystacial pad, and iii) those innervating both structures. Extracellular recordings made during spontaneous movements of the macrovibrisae showed that whisking neurons similar to those observed in the trigeminal ganglion were located in the TMnu. These neurons had different patterns of activation, which were dependent on the type of spontaneous macrovibrissae movement. In particular, their spiking activity tonically increased during fan-like movements of the vibrissae and showed phasic bursting during rhythmic whisking. Furthermore, the same neurons may also respond to masseter muscle stretch. CONCLUSIONS: results strongly support the hypothesis that the TMnu also contains first-order neurons specialized for relaying spatial information related to whisker movement and location to trigeminal-cortical pathways. In fact, the TMnu projects to second-order trigeminal neurons, thus allowing the rat brain to deduce higher-order information regarding executed movements of the vibrissae by combining touch information carried by trigeminal ganglion neurons with proprioceptive information carried by mesencephalic neurons.

Neurons within the trigeminal mesencephalic nucleus encode for the kinematic parameters of the whisker pad macrovibrissae.

It has been recently shown in rats that spontaneous movements of whisker pad macrovibrissae elicited evoked responses in the trigeminal mesencephalic nucleus (Me5). In the present study, electrophysiological and neuroanatomical experiments were performed in anesthetized rats to evaluate whether, besides the whisker displacement per se, the Me5 neurons are also involved in encoding the kinematic properties of macrovibrissae movements, and also whether, as reported for the trigeminal ganglion, even within the Me5 nucleus exists a neuroanatomical representation of the whisker pad macrovibrissae. Extracellular electrical activity of single Me5 neurons was recorded before, during, and after mechanical deflection of the ipsilateral whisker pad macrovibrissae in different directions, and with different velocities and amplitudes. In several groups of animals, single or multiple injections of the tracer Dil were performed into the whisker pad of one side, in close proximity to the vibrissae follicles, in order to label the peripheral terminals of the Me5 neurons innervating the macrovibrissae (whisking-neurons), and therefore, the respective perikaria within the nucleus. Results showed that: (1) the whisker pad macrovibrissae were represented in the medial-caudal part of the Me5 nucleus by a single cluster of cells whose number seemed to match that of the macrovibrissae; (2) macrovibrissae mechanical deflection elicited significant responses in the Me5 whisking-neurons, which were related to the direction, amplitude, and frequency of the applied deflection. The specific functional role of Me5 neurons involved in encoding proprioceptive information arising from the macrovibrissae movements is discussed within the framework of the whole trigeminal nuclei activities.

Role of the trigeminal nerve in regrowth of hypoglossal motoneurons after hypoglossal-facial anastomosis.

Conclusion. Functional recovery of facial muscles following hypoglossal-facial anastomosis (HFA) may be dependent not only on sensory information, relayed via the trigeminal nuclei to the hypoglossal nucleus, but also on extratrigeminal fibers, originating from the hypoglossal nucleus that travel in the infraorbital nerve (ION). This fact helps to explain the ability of hypoglossal neurons, after HFA, to induce contractions of muscles originally innervated from other nervous structures. Objective. The aim of the study was to better understand the role of the trigeminal nerve in reinnervation of facial muscles by hypoglossal motoneurons following HFA. Materials and methods. Central afferences of the ION were analyzed in rats by labeling the exposed nerve with horseradish peroxidase (HRP), whereas central organization of the efferent projections to the vibrissal area was analyzed by labeling the whisker pad muscles of the rat with a 5% solution of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil) in N,N-dimethylformamide. Results. The results show that extratrigeminal fibers, originating in the hypoglossal nucleus, travel along the ION. Retrograde tracing applied to ION or injected into the whisker pad showed labeled neurons in the Pr5 nucleus and all Sp5 trigeminal subnuclei. Small labeled neurons (10-15 microm diameter; 10-12 neurons per section), were also found in the hypoglossal nucleus.

Focal and secondarily generalised convulsive status epilepticus induced by thiocolchicoside in the rat.

The objective of this study was to document the convulsant properties of thiocolchicoside in rats, and to characterise the electroclinical pattern of epileptic seizures. Experiments were carried out in three groups of male Wistar rats: in group A, thiocolchicoside was applied topically to the pia, or given by microinjection to the cerebral cortex (2 microg/microl); in group B, the drug was administered parenterally (6 mg/kg) to rats with minimal lesions of the dura and arachnoid membranes; in group C, thiocolchicoside was administered parenterally (up to 12 mg/kg) to intact rats. In all animals, electroclinical activity was continuously monitored for at least 3 hours after thiocolchicoside injection or application. In group A, electrographic and behavioural activity of focal motor seizures occurred in 100% of animals, developing into a focal status epilepticus; in group B, a multifocal epileptic pattern with secondary generalisation, clinically characterised by clonic or tonic-clonic seizures occurred in 100% of animals, until a secondarily generalised convulsive status epilepticus; in group C, none of animals showed either electrographic or behavioural seizure activity. Our study documents that thiocolchicoside has a powerful convulsant activity in the rat, perhaps due to an antagonistic interaction of the compound with a cortical subtype of the GABA(A) receptor.

The vestibulospinal reflex in humans: effects on paraspinal muscle activity.

OBJECTIVE: To elucidate the electromyographic (EMG) patterns elicited in the superficial paraspinal muscles (sacrospinalis, ileocostalis lomborum, longissimus dorsi and splenium cervicis). MATERIAL AND METHODS: In normal subjects adapted to the dark and standing on a force platform, EMG patterns were elicited in the superficial paraspinal muscles by means of unilateral cold caloric stimulation of the labyrinth. RESULTS: Almost concomitantly with the occurrence of postural derangement of the subject's trunk and head, the EMG patterns of the paraspinal muscles on both sides showed significant and consistent changes. In particular, the paraspinal muscles ipsilateral to the stimulated side showed a significant reduction in activity, while the homologous muscles of the contralateral side markedly increased their activity, with a consistent delay in comparison to the contralateral muscles. Moreover, the EMG changes occurred in a hierarchic fashion in the caudal-rostral direction. In fact, the more powerful and longer-lasting effects were consistently observed in the more caudal muscles, i.e. the sacrospinalis and ileocostalis, while the effects on the longissimus dorsi and splenium cervicis muscles were weak and inconsistent. CONCLUSION: These results suggest a possible use for the EMG patterns resulting from unilateral cold caloric stimulation for clinical evaluation of the vestibulospinal reflex.

Involvement of trigeminal mesencephalic nucleus in kinetic encoding of whisker movements.

In previous experiments performed on anaesthetised rats, we demonstrated that whisking neurons responsive to spontaneous movement of the macrovibrissae are located within the trigeminal mesencephalic nucleus (Me5) and that retrograde tracers injected into the mystacial pad of the rat muzzle extensively labelled a number of Me5 neurons. In order to evaluate the electrophysiological characteristics of the Me5-whisker pad neural connection, the present study analysed the Me5 neurons responses to artificial whisking induced by electrical stimulation of the peripheral stump of the facial nerve. Furthermore, an anterograde tracer was injected into the Me5 to identify and localise the peripheral terminals of these neurons in the mystacial structures. The electrophysiological data demonstrated that artificial whisking induced Me5 evoked potentials as well as single and multiunit Me5 neurons responses consistent with a direct connection. Furthermore, the neuroanatomical findings showed that the peripheral terminals of the Me5 stained neurons established direct connections with the upper part of the macrovibrissae, at the conical body level, with fibres spiralling around the circumference of the vibrissae shaft. As for the functional role of this sensory innervation, we speculated that the Me5 neurons are possibly involved in encoding and relaying proprioceptive information related to vibrissae movements to other CNS structures.

Cytochrome P450 17α-hydroxylase/C(17,20)-lyase immunoreactivity and molecular expression in the cerebellar nuclei of adult male rats.

Several probes have been developed to identify steroidogenic activity in the brain of vertebrates. However, the presence of the cytochrome P450 17α-hydroxylase/C(17,20)-lyase (P450C(17)), an enzyme that converts pregnenolone and progesterone into dehydroepiandrosterone and androstenedione, in specific areas of the cerebellum such as the deep cerebellar nuclei, remains virtually unexplored. Using Western blot analysis and immunohistochemistry, we found molecular expression of P450C(17) in the lateral, interposed and medial deep cerebellar nuclei. Moreover, double immunofluorescence procedures enabled localization of P450C(17) mainly in neurons, axons and glutamatergic synapses. Taken together, these data demonstrate the occurrence of P450C(17) in the deep cerebellar nuclei, and enable the chemical characterization of the cells that express the cytochrome.

Ischemic preconditioning of the muscle improves maximal exercise performance but not maximal oxygen uptake in humans.

Brief episodes of nonlethal ischemia, commonly known as "ischemic preconditioning" (IP), are protective against cell injury induced by infarction. Moreover, muscle IP has been found capable of improving exercise performance. The aim of the study was the comparison of standard exercise performances carried out in normal conditions with those carried out following IP, achieved by brief muscle ischemia at rest (RIP) and after exercise (EIP). Seventeen physically active, healthy male subjects performed three incremental, randomly assigned maximal exercise tests on a cycle ergometer up to exhaustion. One was the reference (REF) test, whereas the others were performed after the RIP and EIP sessions. Total exercise time (TET), total work (TW), and maximal power output (W(max)), oxygen uptake (VO(2max)), and pulmonary ventilation (VE(max)) were assessed. Furthermore, impedance cardiography was used to measure maximal heart rate (HR(max)), stroke volume (SV(max)), and cardiac output (CO(max)). A subgroup of volunteers (n = 10) performed all-out tests to assess their anaerobic capacity. We found that both RIP and EIP protocols increased in a similar fashion TET, TW, W(max), VE(max), and HR(max) with respect to the REF test. In particular, W(max) increased by ∼ 4% in both preconditioning procedures. However, preconditioning sessions failed to increase traditionally measured variables such as VO(2max), SV(max,) CO(max), and anaerobic capacity(.) It was concluded that muscle IP improves performance without any difference between RIP and EIP procedures. The mechanism of this effect could be related to changes in fatigue perception.

Primary afferent plasticity following deafferentation of the trigeminal brainstem nuclei in the adult rat.

Alpha-tyrosinated tubulin is a cytoskeletal protein that is involved in axonal growth and is considered a marker of neuronal plasticity in adult mammals. In adult rats, unilateral ablation of the left facial sensorimotor cortical areas induces degeneration of corticotrigeminal projections and marked denervation of the contralateral sensory trigeminal nuclei. Western blotting and real-time-PCR of homogenates of the contralateral trigeminal ganglion (TG) revealed consistent overexpression of growth proteins 15 days after left decortication in comparison with the ipsilateral side. Immunohistochemical analyses indicated marked overexpression of alpha-tyrosinated tubulin in the cells of the ganglion on the right side. Cytoskeletal changes were primarily observed in the small ganglionic neurons. Application of HRP-CT, WGA-HRP, and HRP to infraorbital nerves on both sides 15 days after left decortication showed a significant degree of terminal sprouting and neosynaptogenesis from right primary afferents at the level of the right caudalis and interpolaris trigeminal subnuclei. These observations suggest that the adaptive response of TG neurons to central deafferentation, leading to overcrowding and rearrangement of the trigeminal primary afferent terminals on V spinal subnuclei neurons, could represent the anatomical basis for distortion of facial modalities, perceived as allodynia and hyperalgesia, despite nerve integrity.

Olfactory-hypoglossal connections.

Natural olfactory stimulation with amyl acetate significantly modulates the electrical activity of hypoglossal neurons and the electromyographic responses of the tongue musculature. The aim of the present study was to identify and characterize, using neuroanatomical and neurophysiological approaches, the pathways involved in the transmission of the olfactory information to the hypoglossal nucleus (XIIn). The neuroanatomical findings provided the initial demonstration that olfactory information is conveyed from the olfactory bulb to the hypoglossal nucleus via the interpeduncular nucleus (IPn) by both fast disynaptic and different polysynaptic pathways. The latter, in particular, involve many of the brain structures that process olfactory information. The electrophysiological studies demonstrated that the IPn neurons respond with a variety of patterns to natural stimulation of the olfactory receptors, thus supporting the hypothesis that the IPn is a crucial relay station for the elaboration and transmission of olfactory stimuli to XIIn.