Contraction times of the cat's tongue muscles measured by light reflection. Innervation of individual tongue muscles

The contraction times of the cat's tongue muscles were measured by recording the fine movements of their surfaces and movements of the tongue's surfaces. The recordings were made by means of a commercial, high-sensitive light reflection transducer (Fairchild FPLA 850) which could be operated without any mechanical loading of the tongue. The contraction times of the intrinsic muscles (including pars longitudinalis superior m. hypoglossi and pars longitudinalis inferior m. styloglossi) measured about 22 ms, while the extrinsic muscles were somewhat slower, around 33 ms. The data are considered in the light of the recently reported histochemical composition of these muscles. The study of the hyoglossal nerve supply to individual tongue muscles revealed that, contrary to earlier reports, the medial nerve sends branches to the intrinsic muscles not only at its distal end but during its entire course through the tongue. The transversal and vertical muscles were found to receive numerous fibers innervating small units of their muscle fibers.     

Taste placodes are primary targets of geniculate but not trigeminal sensory axons in mouse developing tongue

Tongue embryonic taste buds begin to differentiate before the onset of gustatory papilla formation in murine. In light of this previous finding, we sought to reexamine the developing sensory innervation as it extends toward the lingual epithelium between E 11.5 and 14.5. Nerve tracings with fluorescent lipophilic dyes followed by confocal microscope examination were used to study the terminal branching of chorda tympani and lingual nerves. At E11.5, we confirmed that the chorda tympani nerve provided for most of the nerve branching in the tongue swellings. At E12.5, we show that the lingual nerve contribution to the overall innervation of the lingual swellings increased to the extent that its ramifications matched those of the chorda tympani nerve. At E13.0, the chorda tympani nerve terminal arborizations appeared more complex than those of the lingual nerve. While the chorda tympani nerve terminal branching appeared close to the lingual epithelium that of the trigeminal nerve remained rather confined to the subepithelial mesenchymal tissue. At E13.5, chorda tympani nerve terminals projected specifically to an ordered set of loci on the tongue dorsum corresponding to the epithelial placodes. In contrast, the lingual nerve terminals remained subepithelial with no branches directed towards the placodes. At E14.5, chorda tympani nerve filopodia first entered the apical epithelium of the developing fungiform papilla. The results suggest that there may be no significant delay between the differentiation of embryonic taste buds and their initial innervation.     

Motor nuclear representation of masticatory muscles in the rat

Representation of the masticatory muscles within the motor trigeminal nucleus was studied in rats by the horseradish peroxidase (HRP) method and the antidromic field potential method. The motor trigeminal nucleus of the rat could be divided cytoarchitecturally into a dorsolateral and a ventromedial division. Within the dorsolateral division, the temporal muscle was represented dorsomedially, the masseter muscle dorsolaterally and laterally, and the lateral and medial pterygoid muscles ventrolaterally. Within the ventromedial division, the anterior digastric muscle was represented dorsomedially and the mylohyoid muscle ventrolaterally. Distribution of antidromic field potentials evoked by stimulation of the mylohyoid and masseteric nerves coincided with the results from the HRP investigation.     

Central projections of trigeminal primary afferents innervating the nasal mucosa: a horseradish peroxidase study in the rat.

The respiratory region of the nasal mucosa is innervated by the ethmoidal nerve. Chemical nociceptive stimulation of this area leads to upper airway reflexes that prevent access of noxious substances to the respiratory tract and the lungs. In the present study we examined the localization of the cell bodies of the respective primary afferent fibres within the trigeminal ganglion, as well as their central projections. In 25 rats a horseradish peroxidase-wheat germ agglutinin gel was applied to the right nasal cavity. The animals were killed after 48-72 h. For visualization of the tracer the tissue was processed according to the tetramethylbenzidine method. In the trigeminal ganglion almost all labelled cell bodies were localized in a medial band immediately caudal to the entrance of the ophthalmomaxillary branch. Transganglionic projections to the trigeminal brainstem nuclear complex were only localized in the superficial laminae of the subnucleus caudalis and in the subnucleus interpolaris, areas known to be involved in processing of nociceptive information. An additional labelled terminal field was observed in the interstitial subnucleus of the nucleus tractus solitarius, which is involved in respiratory control. These results are in favour of the hypothesis that the ethmoidal nerve in rat constitutes the afferent limb of protective upper airway reflexes since it transmits mainly nociceptive information.     

Startle responses measured in muscles innervated by facial and trigeminal nerves show common modulation

Electromyographic (EMG) potentials of several head muscles were recorded simultaneously in freely moving rats with chronically implanted electrodes. The startle responses of m. temporalis, m. levator auris, and m. levator labii superior were compared. All muscles showed a parallel decrease in latency and an increase in response elicitability and amplitude with an increase in stimulus intensity. A significant latency difference of about 1 ms existed between m. levator auris and m. temporalis. The shortest latency of the EMG response in m. levator auris was 5.5 ms (110 dB SPL). A common fluctuation in response amplitude and latency was found in simultaneous recordings of muscles innervated by the facial and trigeminal nerve, respectively. This shows a common modulatory input to the startle pathway to the cranial motor nuclei.     

The somatostatin sst2A receptor in the rat trigeminal ganglion

Immunohistochemistry for the somatostatin sst2A receptor was performed on the rat trigeminal ganglion to know its function in the trigeminal nervous system. The immunoreactivity was detected in 9.4% of primary sensory neurons in the ganglion. These neurons were small to medium-sized (range=106.5-1123.2 microm(2); mean+/-S.D.=506.3+/-213.2 microm(2)) and predominantly located in the rostromedial part of the ophthalmo-maxillary division. They were also immunoreactive for calcitonin gene-related peptide and the vanilloid receptor subtype 1. In addition, 13.7% of trigeminal neurons which were retrogradely traced with fluorogold from the nasal mucosa exhibited sst2A receptor-immmunoreactivity. Trigeminal neurons which innervated the facial skin and tooth pulp were devoid of the immunoreactivity. In the brainstem trigeminal sensory nuclear complex, both the neuronal cell body and the neuropil exhibited sst2A receptor-immunoreactivity in the superficial medullary dorsal horn.The present study indicates that sst2A receptor-immunoreactive trigeminal nociceptors innervate the nasal mucosa. They may project to the superficial laminae of the medullary dorsal horn.     

Innervation of different peptide-containing neurons in the hippocampus by GABAergic septal afferents

The termination pattern of septohippocampal axons visualized by anterograde transport of Phaseolus vulgaris leucoagglutinin was studied in the hippocampal formation in the rat, with special reference to the innervation of neurons immunoreactive for the neuroactive peptides cholecystokinin, somatostatin or vasoactive intestinal polypeptide. The type I, GABAergic, septohippocampal afferents were shown to terminate on neurons immunoreactive for each of the three peptides. The cholecystokinin-like immunoreactive neurons in all regions, and the somatostatin-immunoreactive cells in stratum oriens of CA1 region were the most preferred targets. Cholecystokinin-immunoreactive cells, especially those in the granule cell layer of the dentate gyrus, were often seen to be contacted by type II (presumed cholinergic) axons as well. The somatostatin-immunoreactive cells in the hilus were also innervated by type I septohippocampal axons, although less frequently than those in stratum oriens of the CA1 subfield. Each type of peptidergic neuron received multiple symmetrical synaptic input from the Phaseolus vulgaris leucoagglutinin-labelled septal afferents, as confirmed by correlated electron microscopy. The majority of these neuropeptide-containing cells are known to be GABAergic, and to have distinct input and output relationships. Thus, the present results demonstrate that the GABAergic septohippocampal pathway can control a wide range of putative inhibitory circuits, and thereby influence the pattern of electrical activity in the hippocampal formation.     

Innervation of the levator ani and coccygeus muscles of the female rat

In humans, the pelvic floor skeletal muscles support the viscera. Damage to innervation of these muscles during parturition may contribute to pelvic organ prolapse and urinary incontinence. Unfortunately, animal models that are suitable for studying parturition-induced pelvic floor neuropathy and its treatment are rare. The present study describes the intrapelvic skeletal muscles (i.e., the iliocaudalis, pubocaudalis, and coccygeus) and their innervation in the rat to assess its usefulness as a model for studies of pelvic floor nerve damage and repair. Dissection of rat intrapelvic skeletal muscles demonstrated a general similarity with human pelvic floor muscles. Innervation of the iliocaudalis and pubocaudalis muscles (which together constitute the levator ani muscles) was provided by a nerve (the "levator ani nerve") that entered the pelvic cavity alongside the pelvic nerve, and then branched and penetrated the ventromedial (i.e., intrapelvic) surface of these muscles. Innervation of the rat coccygeus muscle (the "coccygeal nerve") was derived from two adjacent branches of the L6-S1 trunk that penetrated the muscle on its rostral edge. Acetylcholinesterase staining revealed a single motor endplate zone in each muscle, closely adjacent to the point of nerve penetration. Transection of the levator ani or coccygeal nerves (with a 2-week survival time) reduced muscle mass and myocyte diameter in the iliocaudalis and pubocaudalis or coccygeus muscles, respectively. The pudendal nerve did not innervate the intrapelvic skeletal muscles. We conclude that the intrapelvic skeletal muscles in the rat are similar to those described in our previous studies of humans and that they have a distinct innervation with no contribution from the pudendal nerve.     

Organization of neuronal clusters in the mesencephalic trigeminal nucleus of the rat: fluorescent tracing of temporalis and masseteric primary afferents

The organization of neuronal clusters in the rat mesencephalic trigeminal nucleus (Mes V) was analysed using fluorescent tracing techniques. Simultaneous injections of fluorescent compounds (True blue and Diamidino yellow) were made unilaterally in the masseter and temporalis muscle, or in the masseteric and temporalis nerve. Examination of labeled neurons in frozen brainstem sections showed that (1) the arrangement of temporalis and masseteric primary afferent neurons in the mesencephalic trigeminal nucleus is not somatotopic; (2) primary afferent neurons of both muscles are located at all rostrocaudal levels; and (3) clustering of temporalis, of temporalis and masseteric, and of masseretic afferent neurons occurs at all levels throughout the nucleus.     

Cutaneous nerve regeneration in the rat: reinnervation of the denervated skin by regenerative but not by collateral sprouting

The functional regeneration of cutaneous chemosensitive nerves responsible for the initiation of the neurogenic inflammatory response has been studied in rat hind paw skin. The qualitative and quantitative estimation of plasma extravasation induced by mustard oil, a skin irritant which induces an inflammatory response by the neurogenic route, proved to be a reliable and objective method for studying the regeneration of these cutaneous nerves. The results indicate a complete functional reinnervation by chemosensitive sensory nerves of the skin area previously denervated by crushing the corresponding peripheral nerve. It is concluded that these particular unmyelinated sensory nerves regenerate by means of regenerative sprouting but apparently lack the ability for collateral sprouting. The significance of the present findings in relation to the mechanisms of cutaneous nerve regeneration is discussed.     

Innervation of substantia nigra neurons by cholinergic afferents from pedunculopontine nucleus in the rat: neuroanatomical and electrophysiological evidence

Dopaminergic neurons of the substantia nigra pars compacta are excited by nicotine and acetylcholine, and possess both high-affinity nicotine binding sites and intense acetylcholinesterase activity, consistent with a cholinoceptive role. A probable source of cholinergic afferents is the pedunculopontine nucleus, which forms part of a prominent group of cholinergic perikarya located caudal to the substantia nigra in the tegmentum. Although pedunculopontine efferents, many of them cholinergic, project to the substantia nigra pars compacta, it has not been established whether they terminate in this structure. In the first experiment, which combined retrograde tracing with immunohistochemical visualization of cholinergic neurons, cholinergic cells in and around the pedunculopontine nucleus were found to send projections to the substantia nigra. This projection was almost completely ipsilateral. Subsequent experiments employed anaesthetized rats; kainate was microinfused into tegmental sites in order to stimulate local cholinergic perikarya, and concurrently, extracellular recordings were made of single dopaminergic neurons in the substantia nigra. Consistent with our anatomical findings, unilateral microinfusion of kainic acid in or near the pedunculopontine nucleus increased the firing rate of dopaminergic neurons situated remotely in the ipsilateral substantia nigra. The kainate-induced excitation of nigral dopaminergic neurons was dose-related and was prevented by intravenous administration of the centrally-acting nicotinic cholinergic antagonist mecamylamine. These results suggest that cholinergic perikarya in the vicinity of the pedunculopontine tegmental nucleus innervate dopaminergic neurons in the substantia nigra pars compacta via nicotinic receptors.     

A characterisation of spindle afferents from the intertransverse tail muscles in the rat

Summary The conduction velocities of myelinated afferent axons from the intersegmental muscles at the tail-base in rat range from 5.0–43.0 m · sec^–1, and have a bimodal distribution with a boundary between the groups at 25.0 m · sec^–1. Calculations of the dynamic indices of spindle endings, observed when stretching the muscle, show that axons from primary endings conduct at velocities above 28.0 m · sec^–1 and those from secondary endings at below 24.0 m · sec^–1. These characteristics resemble those of sipindles in the rat hind-limb muscles, and do not compare with thsoe in the more distally situated segmental tail muscles.     

Chemosensitivity of fine afferents from rat skin in vitro

1. Properties of sensory receptors with slowly conducting nerve fibers (less than 10 m/s) were studied using a rat skin-saphenous nerve in vitro preparation where receptive fields of identified single units can be isolated and superfused at the corium side with defined chemical solutions. 2. With mechanical search stimuli, 150 slowly adapting units were identified, 88% C-fibers, and the remainder, A delta-fibers. The majority of these units (65%) were categorized as mechano-heat sensitive ("polymodal") with controlled radiant heat stimulation. The remaining units were classified as low- or high-threshold mechanoreceptors according to their von Frey thresholds. 3. Bradykinin (BK), in concentrations of 10(-8) to 10(-4) M, was repeatedly applied for 1 min at 10-min intervals. Fifty-six percent of the polymodal C-fibers responded to BK (up to 10(-5) M), in contrast to 17% of the heat-insensitive units (P less than 0.01). No correlation between BK sensitivity and conduction velocity or von Frey threshold was found. 4. The BK "threshold concentrations" to excite C- and A delta-fibers were about equally distributed over a range from 10(-8) to 10(-5) M. 5. There was a large interindividual variability in pattern and magnitude of the response to BK. Intraindividually, a marked tachyphylaxis upon repeated BK stimulation was observed. 6. In fibers with a slow development of tachyphylaxis, the effects of conditioning application of different chemicals on BK responsiveness were studied. Norepinephrine in 10(-7) M concentration did not produce a significant effect, whereas 10(-5) M and 10(-4) M seemed to increase the BK responses. 7. Prostaglandin E2 (10(-6) M) caused a weak sensitization to BK on average (n.s.), but serotonin (10(-6) M) was clearly effective (P less than 0.05). 8. The strongest sensitization to BK (P = 0.01) resulted from conditioning heat stimulation, which also uncovered a responsiveness in some units initially insensitive to BK. 9. In some experiments the calcium concentration in the superfusate of receptive fields was lowered to 0.3 mM, which induced ongoing activity in C-fibers and markedly increased the BK responses in two polymodal units tested. Increasing the calcium concentration to 3.0 mM reversed these effects. 10. After completing the BK test protocol, polymodal C-fibers were exposed to other chemicals.(ABSTRACT TRUNCATED AT 400 WORDS)     

Distribution of facial motoneurons innervating the common facial muscles of the rabbit and rat

The distribution of the facial neurons that innervate several facial muscles was determined in the rabbit and the rat by examining the retrograde transport of horseradish peroxidase (HRP). The target muscles were musculus levator nasolabialis, m. levator labii superioris, m. zygomaticus, and m. buccinator pars buccalis, as well as m. parietoauricularis and m. depressor anguli oris in the rabbit and m. levator auricularis posterioris in the rat. Localization of the retrogradely labeled neurons within the ipsilateral facial nucleus was confirmed for all facial muscles examined. Our results showed that m. levator nasolabialis was innervated by neurons located in the dorsal subnucleus, while the motoneurons innervating m. buccinator pars buccalis were distributed within the dorsal part of the intermediate subnucleus of the facial nucleus in the both species. Localization of the labeled motoneurons innervating m. zygomaticus and m. levator labii superioris showed the difference in the distribution within the facial nucleus among the species. Neurons innervating m. parietoauricularis and m. levator auricularis posterioris were localized in somewhat different subregions of the medial subnucleus in these species. M. depressor anguli oris was innervated by the neurons distributed within the intermediate subnucleus of the facial nucleus in the rabbit. Thus, our findings revealed that there is species-specific motor innervation pattern in rabbits and rats, despite several movement of the face is supplied by the homologous facial muscles.     

Kainate receptors in primary afferents to the rat gracile nucleus

The expression of metabotropic glutamate receptor 8 (mGluR8) was studied in the rat hippocampus after pilocarpine-induced status epilepticus (APISE) by light immunohistochemistry and immunoelectron microscopy. At 1 day APISE, mGluR8 immunoreactivity was up-regulated in the entire molecular layer of the dentate gyrus. At 7 days APISE, mGluR8 immunoreactive cells began to appear in the stratum lacunosum moleculare of CA1, and by day 31, they were seen in all layers of CA1. By electron microscopy and double labelling study, the mGluR8 immunoreactive cells were identified as astrocytes. The present novel finding of induced expression of mGluR8 in astrocytes APISE suggests that it may be linked to gliosis.     

Delayed but not loss of gliogenesis in Rbpj-deficient trigeminal ganglion

Somatosensory ganglia including dorsal root ganglion (DRG) and trigeminal ganglion (TG) are derived from a common pool of neural crest stem cells (NCCs), and are good systems to study the mechanisms of neurogenesis and gliogenesis. Previous studies have reported that deletion of Rbpj, a critical integrator of activation signals from all Notch receptors, in NCCs and their derived cells resulted in the delayed gliogenesis at early stage and a loss of glial cells at later stage in the DRG. But the phenotypes in the TG have not been described. Here we reported although the gliogenesis was also delayed initially in Rbpj-deficient TG, it was recovered as the development progressed, as shown by the presence of large number of glial cells in the TG at later stages. However, neuronal reduction was observed in Rbpj-deficient TG, which is similar to what observed in Rbpj-deficient DRG. Taken together, our data indicate the function of Rbpj is diversified and context dependent in the gliogenesis of somatosensory ganglia.     

Myosin isoform transitions during development of extra-ocular and masticatory muscles in the fetal rat

Summary The late fetal development of rat extra-ocular and masticatory muscles was examined by myosin immunohistochemistry. The pattern of slow and neonatal myosin isoform expression in primary and secondary myotubes in these muscles was generally similar to that seen by others in limb muscles. We observed a consistent difference between the Sprague-Dawley and Wistar rats in the degree of maturity reached by all muscles studied at a particular age. In both strains, extra-ocular muscles were also about one day in advance of the masticatory muscles. Thus, secondary myotubes were first seen at E17 in Wistar extraocular muscles, at E18 in Sprague-Dawley extra-ocular muscles and Wistar masticatory muscles, and at E19 in Sprague-Dawley masticatory muscles. There was a strikingly early and complete type differentiation of primary myotubes in extraocular muscles, and tonic myosin first appeared before birth in presumptive extrafusal tonic fibres in the orbital layer of the oculorotatory muscles. Throughout the late fetal period, retractor bulbi was composed of fast myotubes only, but these myotubes were not arranged in classical clusters. In the masticatory muscles at E17/E18 some slow primary myotubes started to express tonic myosin, and these presumptive spindle bag2 fibres were located only in regions of the muscles known to contain spindles in the adult. Presumptive bag1 fibres appeared about a day later (initially without tonic myosin), and in the region of the spindle cluster in anterior deep masseter extrafusal secondary myotube production appeared to be suppressed.