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O Isacson

Publications and source records attributed to O Isacson.

At least 109 records · Page 6Linked to original sources
PubMed1989

Presence of Schwann cells in neurodegenerative lesions of the central nervous system.

Ultrastructural analysis of neurodegenerative CNS lesions produced by an excitotoxic substance revealed that the majority of cells ensheathing axons were not oligodendrocytes. By their morphology and the presence of both a basal lamina and collagen fibers they were identified as Schwann cells. The presence of Schwann cells, whose growth-promoting role in the peripheral nervous system has been largely documented, may account for the development of regenerating growth cones which have been observed in the excitotoxically lesioned central nervous system. Further support for this hypothesis came from the analysis of fetal neural transplants implanted into the lesioned area. Schwann cells ensheathing axons were indeed numerous in the neuron-depleted area surrounding the transplants, where neurite outgrowth of graft origin occurred.

Animals↗
PubMed1989

Nerve growth factor receptor immunoreactivity in the rat suprachiasmatic nucleus.

Using a monoclonal antibody, nerve growth factor receptor has been immunohistochemically identified within the suprachiasmatic nucleus of adult rats. Labelling was most intense in the ventral, lateral and caudal portions of the nucleus and appeared primarily associated with fibers and terminals. These were among the most intensely labelled fibers and terminals in the forebrain.

Animals↗
PubMed1989

A primate model of Huntington's disease: cross-species implantation of striatal precursor cells to the excitotoxically lesioned baboon caudate-putamen.

Ibotenic acid was injected unilaterally into the baboon caudate-putamen (CP) to achieve a neural degeneration model in the primate, with a neuropathology similar to Huntington's disease. Four to six weeks later injections of cell suspensions of striatal precursor cells, obtained by dissection of the fetal rat striatal region (13-15 days gestational age), were made into the excitotoxically lesioned CP of 3 baboons immunosuppressed by Cyclosporin A. Morphological analysis indicated that in one of the baboons, which had the largest lesion of the CP and the shortest survival time (6 weeks after implantation), there was a surviving striatal implant. The implanted neurons grew in high densities in cellular aggregates within the host gliotic CP. These neurons had a neuronal size phenotypical for rat striatum, i.e. on average about a 25% smaller neuronal cell diameter than a similar population in the baboon caudate-putamen. Glial-fibrillary-acid-protein immunoreactivity was present on large astrocytes within the striatal implant, with a distinct border towards the lesion-induced astrogliosis of the host. Neuronal markers for acetylcholinesterase and Leu-enkephalin were distributed in a typical patchy manner in the striatal implants along with fiber staining for tyrosine-hydroxylase-like immunoreactivity (TH) possibly derived from afferent host dopaminergic axons. Some of these fibers in the implants came from intrinsic TH-positive neuronal somata, probably of neocortical fetal origin and transiently expressing the enzyme. In conclusion, the results indicate that neuronal replacement can be achieved by cross-species implantation of fetal striatal precursor cells to the previously neuron depleted primate CP under immunosuppression but that the survival and growth of such implants may be variable and subject to unfavourable trophic conditions.

Animals↗
PubMed1989

Connectivity of striatal grafts implanted into the ibotenic acid-lesioned striatum--III. Efferent projecting graft neurons and their relation to host afferents within the grafts.

Efferent projections of intrastriatally implanted striatal neurons have been studied using a combination of anterograde and retrograde axonal tracers. Adult rats subjected to a unilateral ibotenic acid lesion of the head of the caudate putamen received cell suspension grafts obtained from 14 15-day-old striatal primordia. Three and a half to 20 months after transplantation the rats received either intratransplant injections of the anterograde axonal tracer Phaseolus vulgaris leucoagglutinin or injections of fluorescent retrograde tracers. Fluoro-Gold and rhodamine-labelled latex beads, into the host globus pallidus and substantia nigra. Injections of Phaseolus vulgaris leucoagglutinin located entirely within the grafts labelled axons that ramified extensively within the tissue itself, as well as axons that extended caudally, across the graft host border, along the myelinated fascicles of the internal capsule to arborize in the medial parts of the host globus pallidus. A few axons also reached the entopeduncular nucleus. Injections of Fluoro-Gold into the host globus pallidus labelled large numbers of graft neurons, which had a prominent patchy distribution and were most abundant in the caudal portions of the grafts. Clear retrograde labelling was also seen after injection of Fluoro-Gold or rhodamine beads into the host substantia nigra, although the number of labelled graft neurons was 30-50 times lower than that seen after pallidal injections. Combined injections of Fluoro-Gold into the pallidus and rhodamine beads into the nigra showed that the vast majority of cells labelled from the nigra were also labelled by Fluoro-Gold from the pallidus. In some of the grafted and Fluoro-Gold-injected animals, the fetal donor tissue had been labelled with [3H]thymidine prior to transplantation. Many examples of neurons labelled with both [3H]thymidine and Fluoro-Gold were found after tracer injections into the host globus pallidus, and double-labelled neurons were identified also after Fluoro-Gold injections into the host substantia nigra. In several animals retrograde tracing was combined with labelling of host dopaminergic afferents (by tyrosine hydroxylase immunohistochemistry) and cortical afferents (by injections of Phaseolus vulgaris leucoagglutinin into the host frontal cortex). Comparison of adjacent sections revealed a striking overlap between the patches of Fluoro-Gold-labelled graft neurons (labelled from the host pallidum) and the dense patches of tyrosine hydroxylase-positive terminals. In addition, many of the Fluoro-Gold-labelled cell patches received a high density of cortical afferents labelled by Phaseolus vulgaris leucoagglutinin.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Loss of transmitter-associated enzyme staining following axotomy does not indicate death of brainstem cholinergic neurons.

To assess the validity of using loss of transmitter-associated enzyme staining to document the death of injured cholinergic neurons, vagal and hypoglossal cholinergic neurons were quantitatively identified at 7 and 28 days following unilateral axotomy using staining procedures for choline acetyltransferase (ChAT) or acetylcholinesterase (AChE), using Nissl stains, and using the retrogradely transported dye, True blue, as an intracellular marker. At 7 days post-axotomy, the transmitter-associated enzymes, ChAT and AChE, had disappeared from over 95% of vagal neurons (P greater than 0.001) and from over 50% of hypoglossal neurons (P greater than 0.001) as compared with the unlesioned control side. At 28 days post-axotomy, ChAT and AChE were still absent from over 70% of vagal neurons (P greater than 0.001) but only from about 15% of hypoglossal neurons as compared with unlesioned control sides. In contrast, no statistically significant losses of either vagal or hypoglossal neurons were found at 7 or 28 days post-axotomy using Nissl stains or detection of retrogradely transported True blue. These findings indicate that transmitter-associated enzyme expression can be regulated independently of neuronal survival following injury. We conclude that absence of transmitter-associated enzyme staining is not an absolute indicator of death of cholinergic neurons.

Acetylcholinesterase↗
PubMed1988

Morphological alteration of thalamic afferents in the excitotoxically lesioned striatum.

Excitotoxic lesions of the neostriatum cause anatomical and biochemical changes resembling those occurring in Huntington's disease. One major characteristic of these lesions is that they acutely spare axons of passage and afferent fibers. However, evidence is accumulating that afferent axonal systems decrease their fiber density in the long-term excitotoxic lesion. Ultrastructural changes of neuron-deprived terminals may also occur. A parallel study considering changes in afferent fibers to the excitotoxically lesioned thalamus showed that, a few weeks after neuron-depletion, specific 'point-to-point' systems formed regenerating axonal growth cone-like structures. The present study used the anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) to determine whether specific thalamostriatal afferents form the same kind of regenerating structures following excitotoxic lesion of their target neurons. Thalamostriatal afferents decreased in density over months after lesion, but some were still labeled as long as 4 months after ibotenic acid injection. Remaining afferents formed axonal growth cone-like structures, identified at both light and electron microscopic levels, similar to those observed in the lesioned thalamus. These results demonstrate that in the striatum as in the thalamus, neuron depletion is followed by a long-term alteration of the morphology of some afferent fibers which form regenerating growth cone-like structures. These results are discussed with regard to the possible functional integration of fetal neurons transplanted into previously excitotoxically lesioned areas.

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PubMed1988

Fetal homotypic transplant in the excitotoxically neuron-depleted thalamus: light microscopy.

One month after an in situ injection of kainic acid into the ventrobasal thalamic complex (VB), the lesioned area is totally depleted of neurons. The present study has been undertaken to determine the cytoarchitecture and connectivity of the nucleus constructed by fetal thalamic neurons implanted into the excitotoxically lesioned area. Adult rats received an injection of kainic acid inducing a total neuronal depletion of the right lateral thalamus (including both the nucleus reticularis thalami and the lateral portion of the ventrobasal complex). One month later, homotypic neurons were taken from the dorsal thalamic primordium of rat embryos (gestational age 15-16 days), dissociated, and injected into the lesioned area as a cell suspension. After 2-4-month survival, the cytoarchitecture of the neonucleus formed by the grafted neurons within the previously neuron-depleted area was analyzed. Additionally, connectivity was analyzed in seven rats in which dorsal column nuclei and/or cortical projections to the area were labeled anterogradely with either 3H-leucine or wheat-germ agglutinin conjugated to HRP, and the animals were perfused and processed following various histological procedures (Nissl staining, autoradiographic processing, and histochemistry for visualization of peroxidase). Fetal neurons grew, differentiated, and progressively occupied the previously neuron-depleted area of the adult host CNS. They organized themselves into a neonucleus with particular cytoarchitectural features including 1) the existence of two concentric zones--a central zone containing neurons and glial cells and a marginal zone only filled with a band of glial cells, 2) an increase in cellular density compared to the intact thalamus, 3) the grouping of neurons in spherical clusters, and 4) apparent polymorphism of neuronal somata. Lemniscal and corticothalamic afferents originating from the host were observed in the neonucleus when the fetal neurons had been implanted correctly into the lesioned area but not when they had been misplaced into either normal thalamic tissue or the internal capsule. The afferents labeled from either the dorsal column nuclei or the somatosensory cortex were, however, less dense in the neonucleus than in the normal thalamus. These results are discussed with regard to the normal cytoarchitecture and connectivity of the ventrobasal complex of the rat thalamus.

Animals↗
PubMed1988

Survival and function of dissociated rat dopamine neurones grafted at different developmental stages or after being cultured in vitro.

The in vitro culture approach was combined with the cell suspension grafting technique to examine whether the maturation of dopamine (DA) neurones in vitro imposed similar limitations on their ability to survive grafting as when they are allowed to develop in situ in the fetus. The functional capacity, survival and growth of DA neurones from 2.5- and 7-day-old cultures, grafted to rats with unilateral 6-hydroxydopamine lesions of the nigrostriatal pathway, was compared with similar grafts freshly prepared from fetal donors of embryonic days 14, 16 and 20. Grafts of freshly dissociated mesencephalic DA neurones, taken from embryonic day 14-16 donors and 2.5-day-old cultures, generally survived well and markedly reduced amphetamine-induced rotational asymmetry in the recipient rats. However, when cultured for 7 days prior to grafting, or when taken from 20-day-old fetuses, the mesencephalic DA neurones survived very poorly and the grafts did not have any functional effects. Plating of aliquots of cell suspension used for grafting indicated that the survival rate of dissociated DA neurones is in the same order of magnitude when grown in vitro (about 2 DA neurones per 1000 cells) as when grafted in vivo to the rat striatum (about 1-5 DA neurones per 1000 cells). When the number of surviving grafted DA neurones was plotted against the behavioural effects of the grafts, a threshold number of around 100-200 DA neurones was found necessary to obtain a marked reduction (greater than 50%) in amphetamine-induced rotational asymmetry. Moreover, the survival of 300-500 DA neurones seemed to produce a 'ceiling effect' beyond which additional surviving DA neurones gave rise to little or no further effect on the amphetamine-induced rotational behaviour.

Animals↗
PubMed1988

Striatal grafts in rats with unilateral neostriatal lesions--I. Ultrastructural evidence of afferent synaptic inputs from the host nigrostriatal pathway.

Tyrosine hydroxylase immunocytochemistry, in combination with Golgi impregnation, has been used to study the dopaminergic afferent input to striatal suspension grafts implanted into the previously ibotenic acid-lesioned striatum in adult recipient rats. The rats were perfused for combined light- and electron microscopy at 10-11 months after transplantation, at the end of a series of behavioural experiments and a study of in vivo GABA release, reported in the two accompanying papers. A tyrosine hydroxylase-positive fibre network occurred within the grafts in all eight specimens analysed. The tyrosine hydroxylase-positive fibres had a distinct "patchy" distribution, throughout the graft tissue, and within these patches the terminal density was similar to that of the normal intact striatum. Ultrastructurally, the tyrosine hydroxylase-positive fibres were seen to make abundant synaptic contacts with neuronal elements within the grafts. As in the normal striatum, they were all of the symmetric type and dendritic shafts and spines were the most usual postsynaptic targets. Sections from three of the grafted animals were taken for combined Golgi-impregnation and immunostaining. Only cells of the medium-sized densely spiny type were impregnated in this material. Six of them, which had portions extending into the immunostained neuropil, were drawn using a camera lucida and processed for electron microscopy. Tyrosine hydroxylase-positive boutons were seen to make symmetrical synaptic contacts onto the shafts and spines of the impregnated dendrites, and in one case also with the perikaryon. The results indicate that the medium-sized densely spiny neuron type (which is a predominant target for the dopaminergic afferents in the normal striatum) is abundant in the grafted tissue, and that these neurons represent a synaptic target also for the tyrosine hydroxylase-positive innervation of the striatal grafts.

Animals↗
PubMed1988

Striatal grafts in rats with unilateral neostriatal lesions--II. In vivo monitoring of GABA release in globus pallidus and substantia nigra.

GABA release was recorded in vivo by push-pull perfusion from the globus pallidus and substantia nigra of control rats, rats with unilateral ibotenic acid lesions of the neostriatum, and rats with embryonic striatal tissue grafts implanted in the lesioned striatum. The lesions reduced baseline levels of GABA release to 5% of control levels in the globus pallidus and to 13% of control levels in the substantia nigra pars reticulata. GABA release was substantially restored in both the globus pallidus and substantia nigra of the grafted rats, to 34 and 60%, respectively. Peripheral injection of the dopaminergic stimulant methamphetamine induced a short (lasting approximately 20 min) 4-5 fold increase in GABA release in the intact globus pallidus and a longer (lasting longer than 80 min) increase in the substantia nigra. The stimulatory effect of methamphetamine on GABA release was completely abolished in both sites by the strial lesions, suggesting that the effect was mediated via a direct or indirect dopaminergic action on striatal output neurons. The grafts reinstated methamphetamine-induced stimulation of GABA release in striatal output targets to a level (as a proportion of baseline) that was similar to that seen in the control rats. The results support the view that activation of the dopaminergic inputs to the striatum is functionally excitatory on the major striatal output projections to the globus pallidus and substantia nigra pars reticulata. The results also support the hypothesis that striatal grafts have the capacity to become functionally incorporated by reciprocal graft-host connections into the neural circuitry of the host brain.

Animals↗
PubMed1988

Striatal grafts in rats with unilateral neostriatal lesions--III. Recovery from dopamine-dependent motor asymmetry and deficits in skilled paw reaching.

This study investigated the functional capacity of intrastriatal grafts of embryonic striatal tissue in rats with unilateral ibotenic acid lesions of the neostriatum. The group of grafted rats was compared with lesion-alone and control groups for motor bias, as assessed by tests of rotation induced by dopaminergic, cholinergic and GABAergic drugs, and of skilled paw reaching. Unilateral striatal lesions induced marked ipsilateral turning to apomorphine and methamphetamine, which was substantially ameliorated in the grafted rats. Atropine induced similar rates of moderate (but non-significant) ipsilateral turning in the lesion and graft groups, whereas muscimol and gamma-acetylenic GABA induced no turning bias in any group. The lesioned rats showed a strong bias in their preference to use the paw ipsilateral to the lesion when reaching for food pellets, and a decline in reaching success with both paws. The grafts did not influence the ipsilateral paw preference in this task, but did provide a substantial improvement in the animal's reaching accuracy and ability to retrieve food with either paw. The results indicate that striatal grafts can provide a substantial amelioration of motor impairments induced by striatal lesions. Moreover they suggest that the graft's influence on the host brain is itself under the functional regulation of an afferent dopaminergic input from the host brain.

Alkynes↗
PubMed1988

Connectivity of striatal grafts implanted into the ibotenic acid-lesioned striatum--I. Subcortical afferents.

Subcortical afferents to transplants of fetal striatal tissue, implanted into the excitotoxically lesioned striatum of adult recipient rats, were studied with retrograde and anterograde axonal tracers and immunohistochemistry. One week after a striatal ibotenic acid lesion, involving most of the head of the caudate-putamen, a suspension of fetal striatal tissue (embryonic day 14-15) was injected into the lesioned area. In one group of rats, the ibotenic acid lesion was preceded (10 days) by large intrastriatal injections of True Blue, with injection sites matching the area to be lesioned. This was done to retrogradely pre-label the host brain afferents to the area of the striatum later to be lesioned and grafted. At 3 or 6 months post-transplantation, small injections (50 nl) of rhodamine-labelled latex beads were made into the striatal grafts. In animals where the injections were confined to the graft, retrogradely labelled host brain neurons were found in the thalamus, the substantia nigra, amygdala and dorsal raphe nucleus. Double-labelling analysis revealed that the vast majority of the rhodamine bead-labelled neurons also contained True Blue, which indicates that the host afferents to the graft, to a large extent, were derived from the neurons which normally project to the area of the caudate-putamen which was lesioned by the ibotenic acid injection. To further substantiate these observations a second group of lesioned and grafted animals received unilateral wheatgerm agglutinin-horseradish peroxidase injections into the ipsilateral host thalamus at 4 months post-transplantation in order to anterogradely label the host thalamostriatal axons. In a third group of animals serotonin immunocytochemistry was performed in order to detect possible afferents from the raphe nuclei. In contrast to the serotonin-containing fibers, which were fairly evenly distributed throughout the graft tissue, the peroxidase-labelled thalamic afferents were most prominent in the peripheral zones of the grafts and they were densely aggregated at the graft-host interface. The combined results provide evidence that the intrastriatal grafts receive afferents from the host substantia nigra, thalamus, amygdala and dorsal raphe nucleus, but with different distributions. The afferents from the substantia nigra, amygdala and raphe nuclei seem to distribute throughout the grafted tissue, although they are most dense in the peripheral parts, whereas the thalamic afferents are largely confined to the peripheral areas of the transplants and to the graft-host interface.

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