Distribution and postnatal development of chondroitin sulfate proteoglycans in the perineuronal nets of cholinergic motoneurons innervating extraocular muscles.

Fine control of extraocular muscle fibers derives from two subpopulations of cholinergic motoneurons in the oculomotor-, trochlear- and abducens nuclei. Singly- (SIF) and multiply innervated muscle fibers (MIF) are supplied by the SIF- and MIF motoneurons, respectively, representing different physiological properties and afferentation. SIF motoneurons, as seen in earlier studies, are coated with chondroitin sulfate proteoglycan rich perineuronal nets (PNN), whereas MIF motoneurons lack those. Fine distribution of individual lecticans in the composition of PNNs and adjacent neuropil, as well as the pace of their postnatal accumulation is, however, still unknown. Therefore, the present study aims, by using double immunofluorescent identification and subsequent morphometry, to describe local deposition of lecticans in the perineuronal nets and neuropil of the three eye movement nuclei. In each nucleus PNNs were consequently positive only with WFA and aggrecan reactions, suggesting the dominating role of aggrecan is PNN establishment. Brevican, neurocan and versican however, did not accumulate at all in PNNs but were evenly and moderately present throughout the neuropils. The proportion of PNN bearing motoneurons appeared 76% in oculomotor-, 72.2% in trochlear- and 78.3% in the abducens nucleus. We also identified two morphological subsets of PNNs, the focal and diffuse nets of SIF motoneurons. The process of CSPG accumulation begins just after birth, although considerable PNNs occur at week 1 age around less than half of the motoneurons, which ratio doubles until 2-month age. These findings may be related to the postnatal establishment of the oculokinetic network, performing different repertoires of voluntary eye movements in functionally afoveolate and foveolate animals.

Spinal Excitatory Dynorphinergic Interneurons Contribute to Burn Injury-Induced Nociception Mediated by Phosphorylated Histone 3 at Serine 10 in Rodents.

The phosphorylation of serine 10 in histone 3 (p-S10H3) has recently been demonstrated to participate in spinal nociceptive processing. However, superficial dorsal horn (SDH) neurons involved in p-S10H3-mediated nociception have not been fully characterized. In the present work, we combined immunohistochemistry, in situ hybridization with the retrograde labeling of projection neurons to reveal the subset of dorsal horn neurons presenting an elevated level of p-S10H3 in response to noxious heat (60 °C), causing burn injury. Projection neurons only represented a small percentage (5%) of p-S10H3-positive cells, while the greater part of them belonged to excitatory SDH interneurons. The combined immunolabeling of p-S10H3 with markers of already established interneuronal classes of the SDH revealed that the largest subset of neurons with burn injury-induced p-S10H3 expression was dynorphin immunopositive in mice. Furthermore, the majority of p-S10H3-expressing dynorphinergic neurons proved to be excitatory, as they lacked Pax-2 and showed Lmx1b-immunopositivity. Thus, we showed that neurochemically heterogeneous SDH neurons exhibit the upregulation of p-S10H3 shortly after noxious heat-induced burn injury and consequential tissue damage, and that a dedicated subset of excitatory dynorphinergic neurons is likely a key player in the development of central sensitization via the p-S10H3 mediated pathway.

Pefloxacin induced changes in serotonergic innervation and mast cell number in rat salivary glands.

Pefloxacin is a second-generation fluoroquinolone antibiotic. Besides its advantageous characteristics, side effects including the hypofunction of salivary glands, decreased saliva production, and peripheral neuropathy were observed during the administration of pefloxacin. The aim of this study was to investigate the changes in the number of serotonergic immunoreactive fibers and mast cells after pefloxacin treatment in the parotid and sublingual glands of rats to detect the possible neurotoxic effect of pefloxacin. The adult female rats were treated with intraperitoneal (i.p.) injection of pefloxacin for three or seven days (at a concentration of 20 mg/100g body weight) and the serotonergic innervation pattern along with the change in mast cell number were evaluated by using histochemistry and immunohistochemistry in the parotid and sublingual glands. We found that a three-day treatment significantly increased the number of immunoreactive serotonergic nerve fibers, but after a seven-day treatment the number of serotonin positive nerve fibers decreased almost to values of the control group. The alteration of mast cell number was parallel with the changes of the serotonin positive fibers during the treatment. These results suggest that pefloxacin treatment can modify the finely controlled communication between the immune- and the peripheral nervous systems, resulting neurogenic inflammatory process. The background of this process is the altered serotonergic innervation and the increased number of activated mast cells releasing different mediators for example histamine, which can finally lead to reduced number of serotonin positive nerve fibers after a seven-day treatment of pefloxacin leading to atrophy and hypofunction of the salivary glands.

Brainstem circuits underlying the prey-catching behavior of the frog.

Prey-catching behavior (PCB) of the frog consists of a sequence of movements as a stimulus-response chain of the behavioral pattern in which each action presents a signal for the subsequent event. The transformation of visual information into appropriate spatiotemporal patterns of motor activity is carried out by the motor pattern generators located in the brainstem reticular formation. The motor pattern generators provide input to the motoneurons either directly or via the last-order premotor interneurons (LOPI). Although the feeding program is predetermined in this way, various sensory mechanisms control the motor activity. By using neuronal labeling methods, we have studied the morphological details of sensorimotor integration related to the hypoglossal motoneurons to provide further insight into the neuronal circuits underlying the PCB in ranid frogs. Our major findings are as follows. (1) Dendrodendritic and dendrosomatic contacts established by the crossing dendrites of hypoglossal (XII) motoneurons may serve as a morphological option for co-activation, synchronization and proper timing of the bilateral activity of tongue muscles. The crossing dendrites may also provide a feedforward amplification of various signals to the XII motoneurons. The overlapping dendritic territories of the motoneurons innervating protractor and retractor muscles may facilitate the coordinated activities of the agonistic and antagonistic muscles. (2) The musculotopic organization of the XII motoneurons is reflected in the distribution of LOPI for the protractor and retractor muscles of the tongue. (3) Direct sensory inputs from the trigeminal, vestibular, glossopharyngeal-vagal, hypoglossal and spinal afferent fibers to the XII motoneurons may modulate the basic motor pattern and contribute to the plasticity of neuronal circuits. (4) The electrical couplings observed in the vestibulocerebellar neuronal circuits may synchronize and amplify the afferent signals. The combination of chemical and electrical impulse transmission provides a mechanism by which motoneurons can be activated sequentially.

Musculotopic organization of the motor neurons supplying the mouse hindlimb muscles: a quantitative study using Fluoro-Gold retrograde tracing.

We have mapped the motor neurons (MNs) supplying the major hindlimb muscles of transgenic (C57/BL6J-ChAT-EGFP) and wild-type (C57/BL6J) mice. The fluorescent retrograde tracer Fluoro-Gold was injected into 19 hindlimb muscles. Consecutive transverse spinal cord sections were harvested, the MNs counted, and the MN columns reconstructed in 3D. Three longitudinal MN columns were identified. The dorsolateral column extends from L4 to L6 and consists of MNs innervating the crural muscles and the foot. The ventrolateral column extends from L1 to L6 and accommodates MNs supplying the iliopsoas, gluteal, and quadriceps femoris muscles. The middle part of the ventral horn hosts the central MN column, which extends between L2 and L6 and consists of MNs for the thigh adductor, hamstring, and quadratus femoris muscles. Within these longitudinal columns, the arrangement of the different MN groups reflects their somatotopic organization. MNs innervating muscles developing from the dorsal (e.g., quadriceps) and ventral muscle mass (e.g., hamstring) are situated in the lateral and medial part of the ventral gray, respectively. MN pools belonging to proximal muscles (e.g., quadratus femoris and iliopsoas) are situated ventral to those supplying more distal ones (e.g., plantar muscles). Finally, MNs innervating flexors (e.g., posterior crural muscles) are more medial than those belonging to extensors of the same joint (e.g., anterior crural muscles). These data extend and modify the MN maps in the recently published atlas of the mouse spinal cord and may help when assessing neuronal loss associated with MN diseases.

Projections from the lateral vestibular nucleus to the spinal cord in the mouse.

The present study investigated the projections from the lateral vestibular nucleus (LVe) to the spinal cord using retrograde and anterograde tracers. Retrogradely labeled neurons were found after fluoro-gold injections into both the cervical and lumbar cord, with a smaller number of labeled neurons seen after lumbar cord injections. Labeled neurons in the LVe were found in clusters at caudal levels of the nucleus, and a small gap separated these clusters from labeled neurons in the spinal vestibular nucleus (SpVe). In the anterograde study, BDA-labeled fiber tracts were found in both the ventral and ventrolateral funiculi on the ipsilateral side. These fibers terminated in laminae 6-9. Some fibers were continuous with boutons in contact with motor neurons in both the medial and lateral motor neuron columns. In the lumbar and sacral segments, some collaterals from the ipsilateral vestibulospinal tracts were found on the contralateral side, and these fibers mainly terminated in laminae 6-8. The present study reveals for the first time the fiber terminations of the lateral vestibular nucleus in the mouse spinal cord and therefore enhances future functional studies of the vestibulospinal system.

Musculotopic organization of the motor neurons supplying forelimb and shoulder girdle muscles in the mouse.

We identified the motor neurons (MNs) supplying the shoulder girdle and forelimb muscles in the C57BL/6J mouse spinal cord using Fluoro-Gold retrograde tracer injections. In spinal cord transverse sections from C2 to T2, we observed two MN columns (medial and lateral) both with ventral and dorsal subdivisions. The dorsolateral column consisted of the biceps brachii, forearm extensors, forearm flexors, and hand MNs, and the ventrolateral column consisted of the latissimus dorsi, trapezius, teres major, deltoid, and triceps MNs. The supraspinatus muscle MNs were located in the dorsomedial column, and pectoralis major and serratus anterior MNs were located in the ventromedial columns. MNs of the dorsolateral column innervated the biceps brachii in mid-C4 to mid-C7, forearm extensors in caudal C4 to mid-T1, forearm flexors in rostral C5 to mid-T1, and hand muscles in mid-C8 to mid-T2 segments. The MNs innervating the trapezius were located in mid-C2 to mid-C4, triceps brachii in mid-C6 to rostral T1, deltoid in rostral C4 to mid-C6, teres major in rostral C5 to mid-C8, and latissimus dorsi in mid-C5 to caudal C8. In addition, MNs innervating the supraspinatus were located from rostral C4 to caudal C8, pectoralis major in mid-C6 to mid-T2, and serratus anterior in rostral C5 to caudal C7/rostral C8 segments. While the musculotopic pattern of MN groups was very similar to that documented for other species, we found differences in the position and cranio-caudal extent of some MN pools compared with previous reports. The identification of mouse forelimb MNs can serve as an anatomical reference for studying degenerative MN diseases, spinal cord injury, and developmental gene expression.

Effect of unilateral labyrinthectomy on the molecular composition of perineuronal nets in the lateral vestibular nucleus of the rat.

Disturbances in vestibular functions caused by unilateral labyrinthectomy (UL) are spontaneously restored during the process of vestibular compensation due to the plasticity of CNS. The underlying molecular background of vestibular compensation is not yet fully understood. Recent studies have shown that the extracellular matrix (ECM) molecules have either permissive or non-permissive effect on the neural plasticity. In our previous study we have demonstrated changes in the expression of hyaluronan (HA) in the vestibular nuclei (VN) of the frog following peripheral vestibular lesion. The present work was undertaken to examine the expression of the HA and chondroitin sulfate proteoglycans (CSPGs) in the lateral vestibular nucleus (LVN) of the rat following UL by using histochemical methods. On the first postoperative day, the condensation of the ECM around the neurons, the perineuronal net (PNN) was not distinguished from the surrounding neuropil on the side of UL indicating the desorganization of its molecular structure. At survival day 3, the PNN was recognizable with the HA probe, whereas its staining for the CSPGs was restored by the time of the seventh postoperative day. In the neuropil, the intensity of the HA increased on the operated side, while the CSPGs reaction almost completely disappeared. The present study have demonstrated for the first time that the UL is accompanied by the modification of the HA, and CSPG staining pattern in the PNN of the LVN in the rat. As the reorganization of the PNN corresponds to the restoration of spontaneous activity of vestibular neurons, our study implies the role of HA and CSPGs in the vestibular compensation.

Modification of innervation pattern by fluoroquinolone treatment in the rat salivary glands.

Fluoroquinolone antibiotics (FQAs) are widely used in dental and medical therapy. Despite their known severe adverse actions on the central and peripheral nervous system, little attention has been directed toward the potential toxic side effects of these compounds on the oral tissues. As the saliva secretion is controlled by the nervous system and neuropeptides, the neurotoxic effect of pefloxacin (PEF), a representative member of FQAs, was studied in rats in the present work. Previously, we demonstrated a significant weight loss of parotid gland tissue, a marked decrease in 3H-thymidine incorporation, a decreased volume of saliva and amylase activity of the glandular tissue in response to PEF. Animals received intraperitoneal injection of PEF (20 mg/100 g body weight daily) for 3 and 7 days. Normal histology, and neurofilament 200, substance P (SP) and calcitonin gene-related polypeptide (CGRP) containing nerve fibers were detected with immunohistochemical methods. A marked decrease of the weights in salivary glands and the acinar diameters were measured. Similarly, a strong and significant decrease of the number of SP and CGRP containing nerve fibers were detected. These findings suggest that the impaired morphology and innervation pattern of salivary glands is related to the neurotoxic adverse effect of FQA treatment.

Crossing dendrites of the hypoglossal motoneurons: possible morphological substrate of coordinated and synchronized tongue movements of the frog, Rana esculenta.

Application of different fluorescent tracers to the right and left hypoglossal nerve of the frog revealed the extent of dendrites crossing the midline into the territory of contralateral hypoglossal motoneurons. By using confocal microscopy, a large number of close appositions were detected between hypoglossal motoneurons bilaterally, which formed dendrodendritic and dendrosomatic contacts. The distance between the neighboring profiles suggested close membrane appositions without interposing glial elements. Application of neurobiotin to one hypoglossal nerve resulted in labeling of perikarya exclusively on the ipsilateral side of tracer application, suggesting the absence of dye-coupled connections with contralateral hypoglossal motoneurons. At the ultrastructural level, the dendrodendritic and dendrosomatic contacts did not show any morphological specialization; the long membrane appositions may provide electrotonic interactions between the neighboring profiles. We propose that dendrites of hypoglossal motoneurons that cross the midline subserve one of the morphological substrates of co-activation, synchronization and timing of bilateral activity of tongue muscles during prey-catching behavior of the frog.

Vestibular afferents to the motoneurons of glossopharyngeal and vagus nerves in the frog, Rana esculenta.

The aim of this work was to study whether the vestibular afferent fibers establish direct connections with the motoneurons of glossopharyngeal and vagus nerves of the frog, Rana esculenta. In anaesthetized animals the vestibulocochlear nerve and the common root of glossopharyngeal-vagus and accessory (IX-X-XI) nerves were simultaneously labeled with fluorescein dextran amine (vestibulocochlear nerve) and tetramethylrhodamine dextran amine (IX-X-XI). With a confocal laser scanning microscope we could detect close appositions between the vestibular afferent fibers and somatodendritic components of the general and special visceral motoneurons of the ambiguus nucleus of IX-X nerves. The direct impulse transmission may provide a quick and immediate response of cardiovascular and gastrointestinal system upon body displacement.

Vestibulotrigeminal pathways in the frog, Rana esculenta.

The aim of this study was to investigate whether primary vestibular afferent fibers establish direct connections with the motor and sensory trigeminal system in the brainstem of the frog. The experiments were carried out on Rana esculenta. In anaesthetized animals the trigeminal and vestibular nerves were prepared, and their proximal stumps were labeled either with fluorescein binding dextran amine (trigeminal nerve) or tetramethylrhodamine dextran amine (vestibulocochlear nerve). With a confocal laser scanning microscope we could detect close connections between the vestibular fibers and branches of the dorsal dendritic array of the jaw-closing motoneurons, suggestive of monosynaptic contacts. In the other parts of the brainstem, vestibular terminals were detected in the termination areas of the mesencephalic trigeminal nucleus and of the Gasserian (Vth) ganglion and they were probably involved in polysynaptic connections. In agreement with the results obtained in mammalian species, the present findings suggest that the vestibulotrigeminal relationship is quite complex and uses multiple pathways to connect the vestibular apparatus with the motor and sensory nuclei of the trigeminal nerve in the anurans as well.

Dendrodendritic and dendrosomatic contacts between oculomotor and trochlear motoneurons of the frog, Rana esculenta.

Gaze fixation requires very fast movements of the eye during body displacement. The morphological and physiological background of the very fine and continuous tuning of gaze fixation is not yet fully understood. In a previous study we have shown that the dendrites of oculomotor neurons form bundles which invade the trochlear nucleus, and vice versa, trochlear dendritic bundles invade the oculomotor nucleus. Earlier physiological observations demonstrating electrotonic coupling between dendrites of spinal motoneurons in the frog suggest a similar mechanism between the oculomotor and trochlear motoneurons. We studied a possible morphological basis of gaze fixation. The experiments were carried out on common water frogs, Rana esculenta. The trochlear and oculomotor nerves were cut, and their proximal stumps were labeled simultaneously with different retrograde fluorescent tracers. Using confocal laser scanning microscope we detected a large number of close contacts in both nuclei, the majority of them were dendrodendritic apposition. The distance between the adjacent profiles suggested close membrane appositions without intercalating glial or neuronal elements. At the ultrastructural level, the dendrodendritic and dendrosomatic contacts did not show any morphological specialization; the long membrane appositions may provide ephaptic interactions between the neighboring profiles. This electrotonic coupling between the oculomotor and trochlear nerve motoneurons may promote the co-activation of the muscles responsible for vertical eye movements.

Organization of last-order premotor interneurons related to the protraction of tongue in the frog, Rana esculenta.

Moving visual stimuli elicit a sequence of coordinated activity of muscles including tongue protraction. Morphological and physiological studies fail to reveal any direct tectal projections to hypoglossal motoneurons suggesting that the last-order premotor interneurons (LOPI) are the direct recipients of neural activities generated in the optic tectum. The aim of this study is to analyze the topographical organization of the last-order premotor interneurons related to protractor muscles of the tongue. In Rana esculenta, biotinylated dextran amine (BDA) was injected by iontophoresis into the subnucleus of the hypoglossal nerve containing the motoneurons of protractor muscles of the tongue. For visualizing BDA, sections were treated with avidin-biotin complex and a nickel-enhanced DAB chromogen reaction. The position of labeled neurons was reconstructed with a Neurolucida equipment. Morphologically heterogeneous populations of neurons were detected bilaterally, the majority of them were distributed ipsilateral to the site of injection and extended 1200 microm in rostral and 500 microm in caudal directions. Labeled neurons were found in the rhombencephalic reticular formation, the vestibular nuclei, the nucleus prepositus hypoglossi, the nucleus of solitary tract, the spinal nucleus of trigeminal nerve and the dorsal column nuclei. Our results indicate that the majority of last-order premotor interneurons related to protractor muscles of the tongue are located in the reticular formation of the brainstem. Since this area also receives a significant input from the vestibular system and from proprioceptive fibers, the last-order premotor interneurons presented here may be the target of convergence of sensory modalities involved in prey-catching behavior.

The effect of vestibular nerve section on the expression of the hyaluronan in the frog, Rana esculenta.

Following postganglionic lesion of the eighth cranial nerve, the changes in the expression of hyaluronan (HA), one of the extracellular matrix macromolecules, were examined in the medial (MVN) and lateral (LVN) vestibular nuclei and in the entry or transitional zone (TZ) of the nerve in the frog. HA was detected in different survival times by using a specific biotinylated hyaluronan-binding probe. HA expression was defined by the area-integrated optical density (AIOD), calculated from pixel intensities of digitally captured images. During the first postoperative days the perineuronal net (PN), a HA-rich area around the neurons, was not distinguishable from the surrounding neuropil in the MVN and LVN, characterized by a bilateral drop of AIOD specifically on the operated side. From postoperative day 14 onwards AIOD increased whilst the PN reorganized. In contrast, the AIOD wobbled up and down bilaterally without any trend in the TZ. Statistical analysis indicated that AIOD changes in the structures studied ran parallel bilaterally presumably because of the operation. Our results demonstrated for the first time that (1) the lesion of the eighth cranial nerve is accompanied by the modification of AIOD reflected HA expression in the MVN, LVN and TZ, (2) different tendencies exist in the time course of AIOD in the structures studied and (3) these tendencies are similar on the intact and operated sides. Our findings may suggest an area dependent molecular mechanism of HA in the restoration of vestibular function.

Neurochemical characterization of insulin receptor-expressing primary sensory neurons in wild-type and vanilloid type 1 transient receptor potential receptor knockout mice.

The insulin receptor (IR) is expressed by a subpopulation of primary sensory neurons (PSN), including a proportion of cells expressing the nociceptive transducer vanilloid type 1 transient receptor potential receptor (TRPV1). Recent data suggest functional links between the IR and other receptors, including TRPV1, which could be involved in the development of PSN malfunctions in pathological insulin secretion. Here we used combined immunohistochemical labelling on sections from L4-5 dorsal root ganglia of wild-type (WT) and TRPV1 knockout (KO) mice to examine the neurochemical properties of IR-expressing PSN and the possible effect of deletion of TRPV1 on those characteristics. We found that antibodies raised against the high-molecular-weight neurofilament (NF-200) and the neurofilament protein peripherin distinguished between small and large neurons. We also found that the IR was expressed predominantly by the small peripherin-immunopositive cells both in the WT and in the KO animals. IR expression, however, did not show any preference between the major subpopulations of the small cells, the calcitonin gene-related peptide (CGRP)-expressing and Bandeiraea simplicifolia isolectin B4 (IB4)-binding neurons, either in the WT or in the KO mice. Nevertheless, a significant proportion of the IR-expressing cells also expressed TRPV1. Comparison of the staining pattern of these markers showed no difference between WT and KO animals. These findings indicate that the majority of the IR-expressing PSN are small neurons, which are considered as nociceptive cells. Furthermore, these data show that deletion of the TRPV1 gene does not induce any additional changes in neurochemical phenotype of nociceptive PSN.

Distribution of hyaluronan in the central nervous system of the frog.

The qualitative and quantitative distribution pattern of hyaluronan (HA), a component of the extracellular matrix (ECM), was studied in the frog central nervous system by using a highly specific HA probe and digital image analysis. HA reaction was observed in both the white and the gray matter, showing a very intense staining around the perikarya and dendrites in the perineuronal net (PN). In the telencephalon, strong reaction was found in different parts of the olfactory system, in the pallium, and in the amygdala. In the diencephalon, intensive staining was found in the nucleus of Bellonci, the dorsal habenula, the lateral and central thalamic nuclei, and the subependymal zone of the third ventricle. In the mesencephalon, layers of optic tectum displayed different intensities, with the strongest reaction in layers B, D, F, 3, and 5. Other structures of the mesencephalon showed regional differences. The PN was especially intensively stained around the perikarya of the toral nuclei, the oculomotor and trochlear nuclei, and the basal optic nucleus. In the rhombencephalon, the granular layer of cerebellum, the vestibulocochlear nuclei, the superior olive, the spinal tract of the trigeminal nerve, and parts of the reticular formation showed the most intense reaction in the PN. In the spinal cord, considerable HA staining was found in the white matter and around the perikarya of motoneurons. The present study is the first description of the HA-positive areas of frog brain and spinal cord demonstrating the heterogeneity of HA distribution in the frog central nervous system.

Organization of dye-coupled cerebellar granule cells labeled from afferent vestibular and dorsal root fibers in the frog Rana esculenta.

Application of neurobiotin to the nerves of individual labyrinthine organs and dorsal root fibers of limb-innervating segments of the frog resulted in labeling of granule cells in the cerebellum showing a significant overlap with a partial segregation in the related areas of termination. In different parts of the cerebellum, various combinations of different canal and otolith organ-related granule cells have been discerned. The difference in the extension of territories of vertical canals vs. horizontal canals may reflect their different involvement in the vestibuloocular and vestibulospinal reflex. Dye-coupled cells related to the lagenar and saccular neurons were localized in more rostral parts of the cerebellum, whereas cells of the utricle were represented only in its caudal half. This separation is supportive of the dual function of the lagena and the saccule. The territories of granule cells related to the cervical and lumbar segments of the spinal cord were almost completely separated along the rostrocaudal axis of cerebellum, whereas their territories were almost entirely overlapping in the mediolateral and ventrodorsal directions. The partial overlap of labyrinthine organ-related and dorsal root fiber-related granule cells are suggestive of a convergence of sensory modalities involved in the sense of balance. We propose that the afferent input of vestibular and proprioceptive fibers mediated by gap junctions to the cerebellar granule cells subserve one of the possible morphological correlates of a very rapid modification of the motor activity in the vestibulocerebellospinal neuronal circuit.

Extracellular matrix molecules and their possible roles in the regeneration of frog nervous system.

Recent biochemical and histochemical analyses explored different components of the extracellular matrix (ECM) in the nervous system, and either permissive or non-permissive roles in neuronal development and regeneration were suggested. The aim of this study was to detect the distribution pattern of a few of these molecules in the nervous system of intact frogs and during nerve regeneration. The hyaluronan (HA) and tenascin C reactions were negative in the peripheral nerves, but appeared in their entry zones. In the CNS, different populations of neurons were surrounded with HA and tenascin C-positive material, forming a perineuronal net (PN). The phosphacan reaction was weakly positive in the PNS, and a moderate intensity was detected in the entry zone and in the PN. Laminin and fibronectin immunoreactivity was strong in the PNS, but laminin could not be detected in the CNS. In animals with cut and regenerating vestibulocochlear nerve, the distribution of the ECM molecules in the CNS and PNS characteristically changed from that of the normal pattern. Our results showed a non-homogenous distribution of ECM components in the frog nervous system that could be associated with their different roles in physiological and pathological processes.

Connections of the superior vestibular nucleus with the oculomotor and red nuclei in the rat: an electron microscopic study.

Phaseolus vulgaris leucoagglutinin (PHA-L) was injected into the individual vestibular nuclei of the rat to study their efferent connections. One of the major differences between the connections of these nuclei was found at the level of the mesencephalon: the eye-moving cranial nerve nuclei received the densest projection from the superior vestibular nucleus (SVN). In the present electron microscopic study, we have found that terminals of SVN origin established symmetric synaptic contacts in the oculomotor nucleus. More than two-thirds of PHA-L-labeled boutons terminated on dendrites, the rest of them established axosomatic contacts. Most of the labeled terminals were GABA-positive, supporting the results of previous physiological experiments, which showed inhibitory effects. In the mesencephalon, the other termination area was found in the red nucleus. The PHA-L-labeled boutons of SVN origin were in close contact with the perikarya and proximal dendrites of the magnocellular part of the red nucleus. The types of synaptic contacts and distribution of terminals of SVN origin were similar to those found in the oculomotor nucleus. Our results indicate that the SVN can modify the activity of the cerebellorubral and corticorubral pathways, exerting inhibitory action on the neurons of the red nucleus.