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R Spreafico

Publications and source records attributed to R Spreafico.

At least 91 records · Page 5Linked to original sources
PubMed1993

GABAA receptor impairment in the genetic absence epilepsy rats from Strasbourg (GAERS): an immunocytochemical and receptor binding autoradiographic study.

Some aspects of the GABA and cholinergic systems have been investigated in the cortex and thalamus of GAERS Wistar rats, a model of petit-mal epilepsy, and in a non-epileptic control strain. GABA and its synthetic enzyme, glutamic acid decarboxylase (GAD), were located by immunocytochemistry; the GABAA receptors were evaluated by autoradiography of GABA-enhanced 3H-flunitrazepam binding and by immunocytochemistry using specific antibodies against the beta 2-beta 3 subunits of GABAA receptor protein. GABA and GAD immunocytochemistry did not show up any difference in density or distribution of immunoreactive elements (fibers, terminals and neurons) between epileptic and control animals, but autoradiographic and immunocytochemical studies showed a decreased enhancement of 3H-flunitrazepam binding and of beta 2-beta 3 subunits of GABAA receptor in the sensorimotor cortex and anterior thalamic areas of the epileptic strain. No differences were found in benzodiazepine receptors in the two strains. GABAB receptors were measured as 3H-baclofen binding in a crude synaptic membrane preparation and there was no difference between epileptic and control animals. Choline acetyltransferase, the synthetic enzyme for acetylcholine, and muscarinic receptor subtypes (M1 and M2), visualized respectively by an immunocytochemical procedure and binding autoradiography, did not differ in epileptic and normal rats. The data suggest an impairment of the 'GABAA system' in restricted brain regions of epileptic rats, due to a reduction of receptor beta 2-beta 3 subunits and coupling to benzodiazepine receptors despite the normal synthesis and location of the neurotransmitter.

Acetylcholine↗
PubMed1992

Substance P innervation of the rat and cat thalamus. I. Distribution and relation to ascending spinal pathways.

An antiserum for substance P (SP) with minimal cross-reactivity for other tachykinins was employed to map the distribution of SP-positive nerve fibers and terminals in the thalamus of cats and rats with special emphasis on the innervation by these fibers of nuclei related to the somatosensory system. In both species SP innervation is predominantly along the midline, in medial and posterior thalamic regions, and sparser in sensory relays for specific modalities. Among the most densely innervated nuclei are the parafascicular, paraventricular, rhomboid, central medial and parts of mediodorsal, lateral posterior, and ventral lateral geniculate. SP innervation of somatosensory-related nuclei is also evident in central lateral nucleus, posterior complex (PO), and in ventroposterolateral (VPL) nucleus of both cats and rats. In VPL of cats SP fibers and terminals are present along its ventral and lateral border, a paralaminar area in which spinothalamic fibers have been shown to terminate and where neurons responsive to noxious stimuli have been reported. Also in rats the SP innervation of VPL is similar to that of spinothalamic tract fibers. The SP innervation of somatosensory thalamic nuclei may be supplied, at least in part, by spinothalamic afferent as suggested by the depletion of SP after anterolateral chordotomy but not after ablation of the dorsal column nuclei. The presence of SP-positive spinothalamic neurons in the spinal cord is reported in the following paper.

Amino Acid Sequence↗
PubMed1992

Role of the thalamic reticular nucleus in the generation of rhythmic thalamo-cortical activities subserving spike and waves.

The role of the reticular thalamic nucleus (RTN) in pacing rhythmic cortical activities subserving spike-waves (SW) discharges has been investigated in rats. Intracellular recordings from thalamic slices in vitro demonstrated that RTN neurons from control animals possess a set of Ca2+/K+ membrane conductances which enable them to produce rhythmic oscillatory activities. In vivo, studies of Ca(2+)-conductance blockade by intrathalamic injections of Cd2+ were performed on 24 callosotomized Wistar rats displaying spontaneous SW discharges, bred at the Centre de Neurochimie, Strasbourg. A significant decrement in ipsilateral SW activity was consistently observed in all RTN-injected animals 40 min after Cd2+ injection. By contrast, animals which received Cd2+ injection into the ventroposterior complex (VP) showed only small changes in ipsilateral SW. It is concluded that Ca(2+)-dependent oscillatory properties of the RTN are critical for the expression of genetically determined SW discharges in the Wistar model.

Action Potentials↗
PubMed1992

Distribution of calbindin and parvalbumin in the developing somatosensory cortex and its primordium in the rat: an immunocytochemical study.

Immunocytochemical techniques were used to analyze the distribution of the calcium-binding proteins calbindin and parvalbumin during the pre- and postnatal development of the rat somatosensory cortex. Calbindin occurs in most early differentiated neurons that form the primordial plexiform layer at embryonic day 14. This expression in transient; during the perinatal period, calbindin becomes immunologically undetectable within the structures derived from the primordial plexiform layer, i.e., the prospective layers I and VIb. Immunoreactive neurons are also absent from adult layers I and VIb. Calbindin is also detected in a second population of neurons which, from embryonic day 18 onwards, distributes diffusely within the cortical plate. Some neurons of this population show morphological traits of immaturity, while others show complete dendritic arborization. The definitive pattern of distribution of calbindin-immunoreactive neurons is achieved by postnatal day 22. Infragranular layers contain intensely-immunoreactive cells whose numerical density decreases during postnatal development, whereas in supragranular layers similar neurons are interspersed among numerous faintly-stained neurons. Parvalbumin is detected for the first time at postnatal day 6, within a small group of neurons located in cortical layer V, and extends afterwards through the whole thickness of the cerebral cortex. At this same postnatal stage, groups of immunoreactive puncta are also found in layer IV of the somatosensory cortex; these puncta increase in density progressively and, at embryonic day 13, immunoreactive cells appear also grouped at this level. At this postnatal age, parvalbumin immunostaining delineates the somatosensory map in cortical layer IV. From this stage to adulthood, the number of immunoreactive neurons increases in the whole thickness of the somatosensory cortex. Barrels in layer IV become less distinct as immunoreactive cells and processes invade the septa. Layer IV in the adult somatosensory cortex appears more densely populated by parvalbumin immunoreactive neurons and puncta than in the surrounding areas.

Aging↗
PubMed1992

Prenatal development of calbindin immunoreactivity in the dorsal thalamus of the rat.

The distribution of calbindin immunoreactivity was studied in the developing rat dorsal thalamus at embryonic days 14, 16, 18 and 20. At early stages (days 14-16), calbindin is expressed throughout the dorsal thalamic cell mass. Most intense labeling occurs in cells adjacent to the ventricular surface, in a spatial gradient reflecting the well-known outside-in generation pattern. Between days 16 and 20, calbindin-positive periventricular cells are redistributed in the dorsal thalamus according to two different patterns. They first become oriented tangentially within the periventricular layer, and diminish in number at the central locus where midline thalamic fusion occurs at 18 days. Periventricular calbindin immunoreactivity becomes restricted to a ring of late-born cells surrounding the gray commissure. Recognizable portions of this ring-shaped primordium will mature forming n.paratenialis, n.reuniens, n.paraventricularis, and n.subparafascicularis magnocellularis. Simultaneously, a massive contingent of radially-oriented, fusiform, calbindin-positive young neurons extends from the periventricular ring-shaped aggregate to the lateral brain surface at the caudoventral pole of the dorsal thalamus at embryonic days 17/18. These cells surround the primordium of the medial geniculate body, participating in the constitution of its marginal zone, and invade the lateral posterior nucleus, accumulating within its caudomedial part. Other portions of this stream form the parvocellular subparafascicular nucleus and the peripeduncular nucleus. The observed patterns of calbindin expression suggest that dorsal thalamic postmitotic neurons transiently express the marker during initial phases of axogenesis, whereas a specific, late-born population expresses calbindin continuously into adulthood. This late subpopulation displays migratory behavior, and finally subdivides into several nuclei of the mature midline, superficial and posterior thalamus.

Animals↗
PubMed1991

Postnatal development of calbindin and parvalbumin immunoreactivity in the thalamus of the rat.

The maturation of the calcium binding proteins calbindin-D28k (CB) and parvalbumin (PV) during the first 3 postnatal weeks was studied in the rat thalamus using immunohistochemistry. These two proteins display a non-homogeneous distribution in the adult thalamus. In the rat, CB is mainly localized in the neurons and neuropil of the thalamic midline, intralaminar, and ventromedial nuclei, as well as in the posterior complex. At birth, CB-immunoreactive cell bodies were evident in thalamic midline structures, and especially in the nucleus reuniens. The number of thalamic CB-positive cell bodies, as well as the intensity of the neuropil immunostaining, increased progressively in the first postnatal weeks. This quantitative increase was first apparent in the midline structures and then in the other thalamic territories which are CB-positive in adulthood, and followed a mediolateral gradient. The mature pattern was achieved by the end of the third postnatal week. In the adult rat thalamus the neurons of the reticular nucleus display PV-immunostaining and PV-positive fibers densely innervate most of the dorsal thalamic domains. PV-immunoreactivity was clearly evident at birth in the cell bodies of the reticular nucleus. The density of PV-containing fibers increased progressively after birth in the dorsal thalamus, with a lateromedial gradient. At the end of the third postnatal week the ventroposterior (VP) complex appeared heavily innervated by PV-positive fibers, whose density in more medial structures was still lower than in the adult thalamus. A transient hyperinnervation of PV-immunoreactive fibers, displaying a dishomogenous organization in distinct segments, was observed in VP, and especially in the ventroposteromedial nucleus, during the second postnatal week. Altogether these findings indicate that the maturation of CB and PV requires postnatally a relatively prolonged period of time. The possible involvement of these proteins in different functional aspects of thalamic neuronal maturation is discussed.

Animals↗
PubMed1991

The reticular thalamic nucleus (RTN) of the rat: cytoarchitectural, Golgi, immunocytochemical, and horseradish peroxidase study.

Experiments have been performed on adult albino rats in order to study the cellular organization of the thalamic reticular nucleus. For this purpose four approaches have been used: Nissl stain, Golgi impregnation, retrograde transport of horseradish peroxidase after injection in different thalamic nuclei, and immunocytochemistry with antibodies against GABA and glutamic acid decarboxylase. In sections through the horizontal plane, three morphologically different neurons have been observed. Cells with round perikarya and with multipolar dendrites were found predominantly in the rostral pole of the nucleus. Neurons with large fusiform cell body and with dendrites arborizing mainly on the horizontal plane were detected through the whole extent of the nucleus. Small fusiform neurons were observed almost exclusively in the medial third of the dorso-ventral extent of the nucleus. The Golgi impregnation method demonstrated that dendrites of small fusiform neurons develop in the vertical plane perpendicular to the dendritic arborization of large fusiform neurons. In coronal sections neurons with round perikarya and with large fusiform cell bodies are detectable while small fusiform neurons are only rarely visible. These data have been confirmed by statistical form factor analysis. Moreover, by means of the horseradish peroxidase and the immunocytochemical study, it has been confirmed that all three groups of neurons project within the thalamus and that they are GABAergic. The data concerning the distribution within the nucleus of the three morphologically different neurons are discussed in relation to the topographic distribution of cortical sensory afferents and to the topographic maps within different sectors of the reticular nucleus.

Animals↗
PubMed1991

GABAergic interneurons and neuropil of the intralaminar thalamus: an immunohistochemical study in the rat and the cat, with notes in the monkey.

Immunohistochemistry using antibodies to glutamic acid decarboxylase (GAD) was used to investigate the intralaminar nuclei of the thalamus in rat, cat and monkey. Antibodies to gamma aminobutyric acid (GABA) were also used in the cat. Intralaminar immunoreactive cell bodies were not detected in the rat, but were clearly present in cat and monkey. In the latter species, GABA- or GAD-immunopositive perikarya were distributed throughout the anterior intralaminar nuclei, whereas in the posterior intralaminar complex they prevailed in the lateral part of the centre median nucleus and around the fasciculus retroflexus. Measurements of the area of immunostained intralaminar cell bodies in cat and monkey indicated that they are represented by small neurons. Experiments in the cat, based on retrograde tracers injections involving large sectors of the frontal and parietal cortices and the head of the caudate nucleus, revealed that the GABA- or GAD-immunoreactive cells and the retrogradely labeled projection neurons represented two separate intralaminar cell populations, although the latter also included small cells. Considerable differences were observed in the immunoreactive GABAergic neuropil of the anterior and posterior intralaminar nuclei. Clusters of densely packed bouton-like immunoreactive elements were detected in the former structures in the rat, cat and monkey, and were especially evident in the central lateral nucleus; immunopositive varicose fibers and puncta were diffusely distributed in the posterior intralaminar structures. Taken together with data from the literature, the present findings indicate that in cat and monkey local circuit inhibitory cells regulate not only the activity of principal thalamic nuclei which project densely upon restricted cortical fields, but also of the intralaminar structures which are widely connected with the cerebral cortex and the striatum. Regional variations in the distribution of GABAergic fibers and terminals suggest major differences in the organization of inhibitory circuits and synaptic arrangements of the anterior and posterior intralaminar thalamus.

Animals↗
PubMed1989

Excitatory amino acids mediate responses elicited in vitro by stimulation of cortical afferents to reticularis thalami neurons of the rat.

The effects of the excitatory amino acids on the nucleus reticularis thalami were examined by intracellular recordings from rat thalamic slices. Non-N-methyl-D-aspartate receptor agonists and glutamate induced a membrane depolarization and a reduction in input resistance, while N-methyl-D-aspartate and aspartate induced a prolonged discharge, which in some neurons took the form of a burst firing associated with an apparent increase in membrane input resistance. Both the N-methyl-D-aspartate and the aspartate effects were blocked by D-2-amino-5-phosphonovalerate, while the effects of glutamate, kainate and quisqualate were not. The excitatory postsynaptic potential evoked by corticothalamic fiber stimulation shows two components: an early, short-lasting, 2-amino-5-phosphonovalerate-insensitive portion, and a late, 2-amino-5-phosphonovalerate-sensitive decay phase. It is suggested that glutamate acts in nucleus reticularis thalami cells preferentially on the non-N-methyl-D-aspartate receptors, while aspartate shows an N-methyl-D-aspartate-like effect. The two excitatory amino acids glutamate and aspartate play a determinant role in the modulation of thalamic activity driven by corticothalamic projection.

2-Amino-5-phosphonovalerate↗
PubMed1989

Intrinsic properties of nucleus reticularis thalami neurones of the rat studied in vitro.

1. Neurones of the nucleus reticularis thalami of the rat were studied by intracellular recordings from in vitro slices. The resting membrane potential was -56.28 +/- 5.86 mV (mean value +/- S.D.); input resistance was 43.09 +/- 9.74 M omega; the time constant tau was 16.51 +/- 3.99 ms. At the resting membrane potential tonic firing is present, while at membrane potentials more negative than -60 mV a burst firing mode gradually prevails. 2. Prolonged depolarizing current pulses superimposed on a steady hyperpolarization consistently activated sequences of burst-after-hyperpolarization complexes. The all-or-none burst response consisted of Na+-mediated, TTX-sensitive fast action potentials superimposed on a low threshold spike (LTS). The burst was followed by a stereotyped after-hyperpolarization lasting 100-120 ms (BAHP), with a maxima -85 mV. The BAHP was blocked by Cd2+ and apamine but not by 8-Br cyclic AMP. The early component of BAHP was significantly attenuated by TEA. The oscillatory rhythmic discharges were abolished by agents which blocked the BAHP. 3. The presence of strong after-hyperpolarizing potentials (SAHP and BAHP) in RTN neurones plays a significant role in determining two different functional states, defined as tonic and oscillatory burst firing modes, respectively.

Action Potentials↗
PubMed1988

A comparison of GAD- and GABA-immunoreactive neurons in the first somatosensory area (SI) of the rat cortex.

Neurons immunoreactive for anti-glutamic acid decarboxylase (anti-GAD) and anti-gamma-aminobutyric acid (anti-GABA) were compared in adjacent sections from the rat somatosensory cortex (SI). GAD- and GABA-positive neurons in animals either treated or not treated with colchicine were found to occur at a ratio of 1:2. Measurement of areas of GAD- and GABA-immunoreactive neurons confirmed the presence of an 'exuberant' GABA-positive neuronal population not visualized by the GAD antiserum.

Animals↗
PubMed1988

Electrophysiological characteristics of morphologically identified reticular thalamic neurons from rat slices.

This study is aimed at the investigation of the morphological and electrophysiological characteristics of neurons from the nucleus reticularis thalami in rat thalamic slices incubated in vitro. Ten neurons were recorded in the ventrobasal complex, four of which were successfully injected following horseradish peroxidase injection. Two main types of reticular thalamic neurons were morphologically identified: (1) the small fusiform 'f' cells characterized by a very elongated perikaryon, dendritic arborization prevalent in the rostrocaudal and dorsoventral planes, and an axon without any collaterals branching within the nucleus reticularis thalami; and (2) the large fusiform 'F' neurons with dendrites arborizing mainly in the horizontal plane and with axonal branches within the nucleus reticularis thalami. The electrophysiological properties of the neurons were similar in F and f cells. The reticular neurons showed, in resting conditions, a single spike response followed by a postexcitatory hyperpolarizing potential. The hyperpolarization of these neurons transformed the single spike response into a burst discharge similar to that observed in thalamic relay neurons at resting membrane potential. The same phenomenon was observed when bicuculline was administered by perfusion to the slices and, in this case, a recovery to a single spike response was obtained by a depolarizing d.c. current injection. By contrast, the local administration of GABA induced a depolarization with a pronounced decrease in input resistance. The present data demonstrate the presence of at least two neuronal subtypes within the nucleus reticularis thalami, suggesting that only one is responsible for the phenomenon of auto-inhibition by means of intrinsic axon collaterals. Moreover, it is hypothesized that intranuclear GABAergic collaterals could control neuronal excitability of reticular thalamic cells by both shunting the membrane and shifting the burst firing to a single spike firing mode.

Action Potentials↗
PubMed1988

The intrinsic organization of the ventroposterolateral nucleus and related reticular thalamic nucleus of the rat: a double-labeling ultrastructural investigation with gamma-aminobutyric acid immunogold staining and lectin-conjugated horseradish peroxidase.

An electron-microscopic investigation of the synaptic organization of the rat's ventroposterolateral nucleus (VPL) and of a reticular thalamic nucleus (RTN) area related to somatosensory thalamic nucleus was performed. In a group of 11 rats, wheatgerm agglutinin conjugated to horseradish peroxidase (WGA:HRP) was injected either in the first somatosensory area of cortex (SI) or in the dorsal column nuclei (DCN). The retrogradely and/or anterogradely transported enzyme was visualized using paraphenylenediamine-pyrocatechol (PPD-PC) as substrate. In a second series of six experiments, an immunocytochemical procedure using a specific anti-gamma-aminobutyric acid (anti-GABA) was employed. Postembedding localization of GABA was performed for ultrastructural observation by means of the colloidal gold immunostaining procedure. Thin sections of recognized VPL and RTN areas from WGA:HRP-injected animals were further processed for immunocytochemistry in order to localize simultaneously, at the electron-microscopic level, the transported enzyme and GABA. The results obtained with this procedure demonstrated that HRP-labeled terminals from DCN contacted the soma and proximal dendrites of VPL neurons, while the terminals labeled after SI cortical injections were predominantly localized to the distal portion of the dendrites. The same cortical injection also determined the presence of labeled synaptic boutons contacting the soma, and both proximal and distal dendrites of RTN neurons. GABA-immunolabeled terminals were observed in VPL in a number larger than those observed with other methods, since not only typical F terminals were labeled but also terminals containing round and/or pleomorphic vesicles. GABA-ergic terminals contacted the soma and the proximal and distal dendrites of VPL neurons, while in RTN cells they made synaptic contact mainly with the soma and proximal dendrites. In the double-labeling experiments, terminals containing both HRP and specific immunogold GABA staining were never observed. The present data provide a direct demonstration of the presence of a strong inhibitory input from RTN upon VPL neurons and of the existence of autoinhibition within RTN neurons.

Animals↗
PubMed1987

SII-projecting neurons in the rat thalamus: a single- and double-retrograde-tracing study.

Experiments were performed on adult albino rats, using single-labeling (free horseradish peroxidase [HRP] or wheatgerm agglutinin conjugated to HRP [WGA:HRP]) and double-labeling (fluorescent dyes) techniques to investigate the thalamic projections to the secondary somatosensory cortex (SII) and to demonstrate the presence and location of thalamic neurons projecting to both the primary somatosensory cortex (SI) and SII by way of branching axons. In single-labeling experiments, the tracer was injected in SI or SII with or without electrophysiological control; in double-labeling experiments, fast blue and diamidino yellow were injected into the electrophysiologically identified forelimb areas of SI and SII. Single-tracer experiments showed that after injections in SI, focused in the forelimb representation area, retrogradely labeled neurons were present mainly in the ventral third of the nucleus ventralis posterolateralis (VPL) and in the anterior part of the posterior nuclear complex (PO); labeled neurons were also present consistently in the caudal portion of PO. Injection of tracers in the forelimb or forelimb and hindlimb representation areas of SII resulted in labeling of neurons in the posterior part of PO and in the caudal part of VPL. Double-labeling experiments confirmed the distribution of neurons projecting to SI or to SII, as observed in single-labeling experiments. Some neurons labeled with both tracers were also present. These neurons are interpreted as projecting to both SI and SII by means of axon collaterals and were observed in areas of overlap of the two single-labeled population of neurons--that is, at the border between PO and the ventroposterior complex, and in the medial part of caudal PO. Comparison of these data with those obtained after injections of tracers in SI and SII of cats (Spreafico et al., 1981b) suggests that in both species thalamic neurons projecting to these two areas are largely segregated, though partially overlapping; and that thalamic neurons projecting simultaneously to SI and SII, modest in number in cats, are even sparser in rats.

Afferent Pathways↗
PubMed1986

GABA immunoreactivity in the thalamic reticular nucleus of the rat. A light and electron microscopical study.

The thalamic reticular nucleus (TRN) of the rat has been studied immunocytochemically using an antiserum against the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). Combined light and electron microscopic investigations by means of peroxidase-antiperoxidase and immunogold labeling show that this nucleus contains a homogeneous population of GABA-immunoreactive neurons receiving extensive GABAergic connections suggestive of self-inhibitory inputs.

Animals↗
PubMed1986

GABAergic neurons are present in the dorsal column nuclei but not in the ventroposterior complex of rats.

Neurons containing glutamatic acid decarboxylase (GAD) are known to exist in the spinal dorsal horn, dorsal column nuclei (DCN), n. ventralis posterior (VP), and somatosensory cortex of cats. Recent work suggested that species differences exist concerning the presence and/or density of GAD-positive neurons in VP. The present experiments demonstrate that, in contrast with carnivores and primates, the rat's VP contains virtually no GAD-positive neurons and that virtually all neurons in it project to the cortex. This conclusion is supported by the failure to find, in Golgi-impregnated material, neurons with characteristics commonly attributed to Golgi type II neurons in VP of cats. The lack of GAD-positive neurons in VP of rats contrasts also with the presence of such neurons in the DCN in the same species. As in cats, about one third of the neurons in the cuneate n. are GAD-positive; these have mostly small perikarya and they are present throughout the nucleus. It is likely that these are intrinsic neurons, i.e. non-projecting beyond the limits of the DCN since a comparable percentage of neurons are unlabeled by simultaneous injections of horseradish peroxidase in multiple targets of the DCN. Like GAD-positive neurons, neurons unlabeled by the retrograde transport of HRP have, for the most part, small perikarya. It is possible that inhibitory mechanisms necessary for basic transfer functions in VP of rats are sustained through projections to this nucleus from the n. reticularis thalami. Extrinsic source of GABAergic input to the DCN seem to be absent or very weak. From this and previous evidence it may be proposed that intrinsic inhibitory interneurons have gradually developed in VP of rabbits, carnivores, and primates in parallel with more elaborate levels of thalamic integration of somatosensation.

Animals↗