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Biomedical subjects

M R Pantò

Publications and source records attributed to M R Pantò.

16 recordsLinked to original sources

Expression of pannexin1 in the CNS of adult mouse: cellular localization and effect of 4-aminopyridine-induced seizures.

The expression pattern of pannexin1, a gene coding for a protein that forms gap junction channels, was studied as both mRNA and protein in the CNS of adult mouse. Pannexin1 was widely expressed in the CNS by neuronal cell types but not glial cells, except for Bergmann glial cells of the cerebellar cortex. Cells positive to Ca-binding proteins, principally parvalbumin, but also calbindin and calretinin, as well as glutamate decarboxylase 67 kDa isoform, were pannexin1-positive. Pannexin1 labeling was found in cells which are known to exhibit spontaneous and synchronous discharge, such as neurons of the inferior olivary complex and the reticular thalamic nucleus, and also in neurons whose electrical activity is not coupled with neighboring cells, such as motoneurons of the spinal cord. The analysis of cellular localization showed puncta that surrounded cell bodies (e.g. the pyramidal cells of hippocampus) or restricted areas inside the cell bodies (e.g. the spinal motoneurons). In Bergmann glial cells the staining was present as fine grains that covered a large part of the cellular surface. Pannexin1 stained cells that previous studies have reported as expressing connexin36, another protein forming gap junction channels. Thus, it was possible that these two proteins could be integrated in the same functions. Since connexin36 expression levels change after seizures, we examined the expression of both pannexin1 and connexin36 in cerebral cortex, hippocampus, cerebellum and brain stem at different time intervals (2, 4 and 8 h) after i.p. injection of 4-aminopyridine, which resulted in systemic seizures. The only modification of the expression levels observed in this study concerned the progressive decrement of the connexin36 in the hippocampus, while pannexin1 expression was unchanged. This finding suggested that pannexin1 and connexin36 are involved in different functional roles or that they are expressed in different cell types and that only those expressing the Cx36 are induced to apoptosis by epileptic seizures.

4-Aminopyridine↗

Laterality of the pontocerebellar projections in the rat.

This study aimed to investigate the trajectory of fibres from the pontine nuclei that reach the two sides of the cerebellum. Injections of biotinylated dextran amine (BDA) were made within the basilar pontine nuclei (BPN) and the nucleus reticularis tegmenti pontis (NRTP) in one side of rats with electrolytic injury of the middle cerebellar peduncle (MCP), ipsilateral or contralateral to the side of injection. Fibres were traced from the pontine nuclei (BPN and NRTP) to both sides of the cerebellum passing through the respective MCPs. The study carried out in rats with injury to one peduncle showed projections segregated to the half-side of the cerebellum innervated by the intact peduncle. The laterality observed was confirmed by a retrograde tracer study. In fact, injections of different fluorescent tracers in rats with injury of single MCP showed that in the pontine nuclei only cell bodies stained by the tracer injected in the half-cerebellum ipsilateral to the intact peduncle. Finally, similar injections (i.e. different fluorescent tracers in symmetric areas of the cerebellar cortex) in the cerebellum of intact brain rats showed that BPN and NRTP differ for the laterality of their projections. In fact, 82% of BPN cells project contralaterally and 18% ipsilaterally, whereas 60% of NRTP cells project contralaterally and 40% ipsilaterally. In conclusion, this study showed that the MCPs receive fibres from the pontine nuclei of both sides and project to the ipsilateral half of the cerebellum and that different contingents of projections to the two sides of the cerebellum arise from BPN and NRTP.

Animals↗

Multiple zonal projections of the nucleus reticularis tegmenti pontis to the cerebellar cortex of the rat.

Compartmentalization (alternating labelled and unlabelled stripes) of mossy fibre terminals was found in the cerebellar cortex after iontophoretic injections of biotinylated dextran amine into discrete regions of the nucleus reticularis tegmenti pontis (NRTP). The zonal pattern was only observed when volumes of nuclear tissue ranging from 4.5 x 106 to 17.66 x 106 microm3 were impregnated. Up to nine compartments (i.e. up to five stripes separated by four interstripes) were found in crus I and in vermal lobule VI. Up to seven compartments (four stripes and three interstripes) were found in crus II; up to five compartments (three stripes and two interstripes) were identified in the lobulus simplex, the paraflocculus and vermal lobules IV, V and VII; up to three compartments (two stripes and one interstripe) were identified in the paramedian lobule and, finally, up to two compartments (one stripe and one interstripe) were identified in the copula pyramidis, in the flocculus and in vermal lobules II, III, VIII and IX. The projections of the NRTP are arranged according to a divergent/convergent projection pattern. From single injections in the NRTP, projections were traced to a set of cortical stripes widely distributed over the cerebellar cortex. The set of stripes labelled from different regions of the NRTP partially overlapped but complete overlap was never found. This finding revealed that the topographic combination of the projections of the NRTP to the cerebellar cortex is specific for each region of the NRTP. Finally, the projections to single cortical areas were arranged according to a pattern of compartmentalization that is specific for each cortical area, independent of the site of injection in the NRTP and of the number of stripes evident in the cortex.

Animals↗

Corticonuclear projections of the cerebellum preserve both anteroposterior and mediolateral pairing patterns.

The aim of the present study was to establish whether a diverging arrangement of the corticonuclear cerebellar projections exists and, if so, what relation it has with the inferior olivary complex. Iontophoretic injections of a 1 : 1 mixture of tetramethylrhodamine dextran amine and biotinylated dextran amine into the cerebellar cortex orthogradely labelled fibre terminals in the cerebellar nuclei and retrogradely labelled cell bodies in the inferior olivary complex. The injections were into A, B, C2, C3, D1 and D2 bands. These injections showed diverging projections to the cerebellar nuclei, with 'primary projections' directed to the nuclear region previously reported to be specifically connected with the injected band and 'secondary projections' directed to other nuclear regions. Secondary projections from the A, C2 and C3 bands diverged to nuclear regions primarily controlled by cortical bands lateral to those injected. Secondary projections from the D1, and D2 bands diverged to nuclear regions primarily controlled by cortical bands medial to those injected. Moreover, injections distributed along the D1 and D2 bands showed similar sets of nuclear targets, while those distributed along the A, C2 and C3 bands showed two sets of nuclear targets in relation to the anteroposterior location of the injected area within these bands. The cortical areas that projected to the same set of nuclear targets were innervated from single olivary regions, while those that projected to different sets of nuclear targets were innervated from different subsets of single regions of the inferior olive. The results suggest that the olivary bands of the cerebellar cortex project to the cerebellar nuclei with a diverging pattern that is organized in both the mediolateral and the anteroposterior axes.

Animals↗

Diverging projections of the C2 and D2 olivocorticonuclear cerebellar pathways of the rat.

A divergent mediolateral projection to the cerebellar nuclei of the C2 and the D2 olivocorticonuclear cerebellar pathways was found after segregate injections of a tracer (either WGA-HRP or FR or BDA) in the rostral (D2 area) or caudal side (C2 area) of the rat paraflocculus. The C2 olivary area of the cerebellar cortex sends most of its nuclear projection to the nucleus interpositus posterior (classically perceived as the nuclear target of the C2 olivocorticocerebellar pathway) and a smaller contingent of fibres to the parvocellular region of the nucleus lateralis (classically perceived as the nuclear target of the D2 olivocorticocerebellar pathway). The D2 olivary area of the cerebellar cortex sends most of its nuclear projection to the parvocellular region of the nucleus lateralis (classically perceived as the nuclear target of the D2 olivocorticocerebellar pathway) and a smaller contingent of fibres to the magnocellular region of the nucleus lateralis (classically perceived as the nuclear target of the D1 olivocorticocerebellar pathway). The lateral interaction of the D2 and the C2 olivocerebellar pathways could represent the anatomical substrate for the functional integration of different olivocerebellar compartments.

Animals↗

The projections of the lateral reticular nucleus to the deep cerebellar nuclei. An experimental analysis in the rat.

The projections of the lateral reticular nucleus (LRN) to the cerebellar nuclei were studied using the retrograde axonal transport of tetramethyl rhodamine dextran amine (10% solution in 0.01 M neutral phosphate buffer) in 19 adult Wistar strain rats. The cerebellar nuclei receive topographically organized projections from the LRN. The projections are bilateral with an ipsilateral predominance and they are symmetrical. The contralateral component is progressively larger for projections to the nuclei interpositalis, to the nucleus lateralis and to the nucleus medialis. The projections to the various cerebellar nuclei arise from rostrocaudally oriented columns of neurons located in different (partly overlapping) areas of the magnocellular division of the LRN. The nucleus lateralis receives terminals from the dorsomedial area (mainly from the rostral level of the LRN), the nuclei interpositalis from the dorsolateral area (mainly from the central level) and the nucleus medialis from the intermedioventral area (mainly from the caudal level). Afferent fibres from the small subtrigeminal division were traced to the three cerebellar nuclei and from the parvocellular division to the nuclei interpositalis and medialis. The density of the projections from the LRN to the nuclei interpositalis increases progressively with the shift of the terminal field from the rostrolateral to the caudomedial part of the nucleus. The projections to the nucleus lateralis reach principally the dorsolateral hump, whereas only a few neurons project to the other divisions (parvo- and magnocellular). The projections to the various regions of the nucleus medialis show different densities. The highest density was found for projections to the caudal part, in particular to the dorsolateral protuberance and to the ventrolateral area of the middle division. Conversely, a low density of projections was found for the other areas of the middle division. The regions of the magnocellular division of the LRN which project to the nuclei lateralis (and are thus related to the cerebral cortex), interpositalis (related to the red nucleus) and medialis (related to the spinal cord) also receive afferent terminals from the cerebral cortex, the red nucleus and the spinal cord respectively, in addition to various afferent inputs. Thus, each of these areas is apparently concerned with integrating some spinal and supraspinal information in reverberating circuits.

Afferent Pathways↗

An autoradiographic study of the cerebellopontine projections from the interposed and lateral cerebellar nuclei in the rat.

The projections from the cerebellar lateral and interposed nuclei onto the basilar pontine gray and nucleus reticularis tegmenti pontis (NRTP) have been studied in the rat by the use of the autoradiographic technique. Projections from both nuclei are mainly contralateral. Fibers from the lateral nucleus cover most of the NRTP, except its medial, parvocellular portion. In the pontine gray proper, fibers from the lateral nucleus are distributed to three rostrocaudally oriented columns: (i) in the medial nucleus, (ii) in the ventral nucleus, and (iii) in the dorsolateral and lateral nuclei. The projection is topographically arranged, so that caudal parts of the lateral cerebellar nucleus tend to project to more rostral regions than rostral parts of the lateral nucleus. The interposed nucleus gives rise to a sparser projection, apparently limited to the ventral NRTP and immediate peripeduncular zones. The functional implications of these results are discussed, with particular emphasis on the convergence of corticopontine afferents to the pontine regions involved, and on the reciprocal pontocerebellar pathways from these same regions.

Animals↗

[Ponto-medullary projections to the spinal cord: a quantitative study in the cat].

The purpose of this research has been to study quantitatively the ponto-medullary projections to the lateral and ventral funiculi in both cervical (C3) and thoracic (Th8) levels. The method consisted in the partial interruption of the spinal cord, sparing one funiculus in which 5-30 microliter of HRP 30% solution was injected at about 1 cm caudally. 19 cats were utilized, 12 for cervical (6 for lateral and 6 for ventral funiculus) and 7 for lumbar injection (3 for lateral and 4 for ventral funiculus). The structures which project to both funiculi of spinal cord (RM, RL, Poo, PR at Th8 level; RM, RL, Poo at C3 level) could exert integrative effects on the proximal and distal segments of the limbs. The structures which project only to FV (VL and Poc at Th8 level; VL, VIN and Poc at C3 level) or to FL (LSC at Th8; PR at C3) could be implied in motor control of only the proximal or distal regions.

Animals↗

An autoradiographic study of the cerebellopontine projections in the rat. I. Projections from the medial cerebellar nucleus.

The medial cerebellar nucleus of the rat is shown by the autoradiographic technique to project to both the contralateral nucleus reticularis tegmenti pontis and the pontine nuclei proper. The former projection is more concentrated in the medial--parvocellular --region. In the pontine gray, the bulk of the projection concerns the dorsal aspect of the medial nucleus. Rostral parts of the medial cerebellar nucleus project to caudal pontine levels whereas caudal parts seem to project throughout the rostrocaudal extent of the basilar pons.

Animals↗

[Convergence effects evoked in neurons of the thalamic ventrolateral (VL) nucleus by the stimulation of paleo- and neocerebellar nuclear efferents].

The functional characteristics of the cerebellar nuclear outputs (fastigial, F.; interpositus, I.; dentate, D. nuclei) convergence on single neurones of th ventrolateral (VL) thalamic nucleus were studied in chloralosed, curarized and artificially ventilated cats (pCO2 continuously monitored), by means of the usual electrophysiological stimulation and intracellular recording techniques. From the experimental results on 90 VL n. neurones, with the analysis of the evoked cerebellar nuclear outputs unitary responses, it is possible to conclude: a) 69% of the reactive VL n. units are convergence neurones (CN); b) 34% of the CN are reactive to stimulation of all three cerebellar nuclei, showing similar patterns, with predominantly excitatory effects when located in the dorso-medial VL n. region and opposite patterns (excitation and inhibition) when located in the ventro-lateral VL n. region; c) 30% of the CN are reactive to the F.n and I.n. stimulation, often with opposite patterns; d) 36% of the CN are reactive to the D.n.-F.n. and D.n.--I.n. stimulation, usually with similar patterns. From this evidence, the effects evoked by the D.n. stimulation on to VL n. neurones seem to potentiate the F.n. and I.n. actions, whereas teh convergent F.n.-I.n. inputs exert often opposite effects on the same neurone. The dorso-medial and the ventro--lateral VL n. regions also seem to receive different actions from the converging cerebellar nuclear outputs. All the results are functionally discussed in terms of a possible integration between posture and voluntary movements at the level of the thalamic relay nucleus of the cerebello-cortical pathway.

Animals↗

Single muscle organization of interposito-rubral projections.

In unanesthetized neuraxis intact cats microstimulation of the interpositus nucleus (IN) which activated a single flexor or extensor muscle in limbs, was used to investigate changes of unitary discharged of rubrospinal (RST) cells. Recordings were made from sites the stimulation of which excited the same muscle activated by the IN (agonist cells), its antagonist (antagonist cells) or heteronymous muscles (heteronymous cells). Cats submitted to chronic cerebellar decortication, acute brachium conjunctivum (BC) section, acute prerubral hemidecerebration or chronic prerubral hemidecerebration and contralateral BC section, were used as controls. It was shown that agonist RST cells were monosynaptically fired from IN, while antagonist cells were inhibited and the heteronymous ones were not influenced. Cerebellar efferents within the BC mediate both excitatory and inhibitory effects, but cerebellar cortex and prerubral structures were not involved in their production.

Animals↗