PubMed Health⌕ Search

Biomedical subjects

G Raisman

Publications and source records attributed to G Raisman.

At least 19 recordsLinked to original sources
PubMed2002

Fluorescent retrograde neuronal tracers that label the rat facial nucleus: a comparison of Fast Blue, Fluoro-ruby, Fluoro-emerald, Fluoro-Gold and DiI.

Many fluorescent retrograde tracers are commercially available for neuroanatomical studies. They have been used with varying success in different models and can be very effective in the study of the facial nerve and nucleus. We compare the tracers Fast Blue (FB), Fluoro-ruby, Fluoro-emerald, Fluoro-Gold (FG), and DiI in the rat facial nucleus after application to the buccal division of the nerve. There were no significant differences between counts of cells on the left and right sides of the brain stem with any of the tracers. FB produced a lower sample variation than the other tracers, and together with DiI, demonstrated greater axonal labelling when applied to the surface of the epineurium. FB and FG resulted in strong retrograde labelling of the facial nucleus after only 2 days from injection. All the tracers produced adequate cell labelling after 1 week from nerve application. This labelling persisted for up to 8 weeks for most tracers except FG, which did not produce satisfactory labelling at 8 weeks.

Amidines↗
PubMed2002

How do transplanted olfactory ensheathing cells restore function?

In this article, we review our work on regeneration of the corticospinal tract in rats following a lesion at upper cervical level. We outline the rationale for using olfactory ensheathing cells, and summarize the evidence for regeneration and functional recovery. The present interpretation on the mechanisms of functional recovery is partly hypothetical, and we emphasize where further experimental evidence is needed.

Animals↗
PubMed2001

Columns of Schwann cells extruded into the CNS induce in-growth of astrocytes to form organized new glial pathways.

Our previous work showed that stereotaxic microextrusion of columns of purified peripheral nerve-derived Schwann cells into the thalamus of syngeneic adult rats induces host axons to grow into the column and form a new fiber tract. Here we describe the time course of cellular events that lead to the formation of this new tract. At 2 h postoperation, numerous OX42-positive microglia accumulated at the graft-host interface, after which donor columns became progressively and heavily infiltrated by microglia/macrophages that took on an elongated morphology in parallel with the highly orientated processes of the donor Schwann cells. The penetration of host astrocytic processes into the Schwann cell columns was substantially slower in onset, being first detected at 4 days postoperation. This event was contemporaneous with the in-growth of host thalamic axons. Between 7 and 14 days postoperation, GFAP-positive astrocytes became fully incorporated into the transplants, where they too adopted an elongated form, orientated in parallel with the longitudinal axis of the graft. Thus, the columns became a mosaic of elongated and highly orientated donor Schwann cells intimately mingled with host microglia, astrocytes, and numerous, largely unbranched 200-kDa neurofilament-positive axons from the adjacent thalamus. Electron microscopy demonstrated that the processes of donor Schwann cells and host astrocytes within the column formed tightly packed bundles that were surrounded by a partial or complete basal lamina. Control columns, formed by extruding freeze-thaw-killed Schwann cells or purified peripheral nerve fibroblasts induced a reactive injury response by the adjacent host microglia and astrocytes, but neither host astrocytes nor neurofilament-positive axons were incorporated into the columns. A better understanding of the mechanisms that regulate the interactions between donor and host glia should facilitate improved integration of such grafts and enhance their potential for inducing tissue repair.

Animals↗
PubMed2001

Olfactory ensheathing cells - another miracle cure for spinal cord injury?

Several recent publications describe remarkably promising effects of transplanting olfactory ensheathing cells as a potential future method to repair human spinal cord injuries. But why were cells from the nose transplanted into the spinal cord? What are olfactory ensheathing cells, and how might they produce these beneficial effects? And more generally, what do we mean by spinal cord injury? To what extent can we compare repair in an animal to repair in a human?

Animals↗
PubMed2000

Improved post-embedding immunocytochemistry of myelinated nervous tissue for electron microscopy.

The particularly high lipid content of normal mature adult myelin sheaths, together with the light fixation protocols usually necessary to retain antigenicity, combine to make white matter nervous tissue an especially problematical subject for post-embedding immuno-electron microscopy using modern acrylic resins. Fixation and infiltration modifications to standard processing schedules for Lowicryl were found to greatly improve the embedding and therefore the resulting morphology. This in turn improved the signal to noise ratio by reducing the high non-specific backgrounds usually found in poorly infiltrated areas. Using Lowicryl HM20, we have been able to obtain satisfactory immunostaining for myelin basic protein with good retention of structural integrity in the myelin of both normal and lesioned adult cortico spinal tract.

Animals↗
PubMed2000

Repair of corticospinal axons by transplantation of olfactory ensheathing cells.

This paper examines the possibility of repairing cut central connections by transplantation of glial cells which modify the glial pathways and take advantage of the inherent growth capacity in adult neurons. We found that transplants of cultured olfactory ensheathing cells into lesions of the adult rat corticospinal tract induced long growth of cut axons across the lesion. Acquisition of a directed forepaw reaching function was restored on the operated side.

Animals↗
PubMed1999

Signalling by the RET receptor tyrosine kinase and its role in the development of the mammalian enteric nervous system.

RET is a member of the receptor tyrosine kinase (RTK) superfamily, which can transduce signalling by glial cell line-derived neurotrophic factor (GDNF) and neurturin (NTN) in cultured cells. In order to determine whether in addition to being sufficient, RET is also necessary for signalling by these growth factors, we studied the response to GDNF and NTN of primary neuronal cultures (peripheral sensory and central dopaminergic neurons) derived from wild-type and RET-deficient mice. Our experiments show that absence of a functional RET receptor abrogates the biological responses of neuronal cells to both GDNF and NTN. Despite the established role of the RET signal transduction pathway in the development of the mammalian enteric nervous system (ENS), very little is known regarding its cellular mechanism(s) of action. Here, we have studied the effects of GDNF and NTN on cultures of neural crest (NC)-derived cells isolated from the gut of rat embryos. Our findings suggest that GDNF and NTN promote the survival of enteric neurons as well as the survival, proliferation and differentiation of multipotential ENS progenitors present in the gut of E12.5-13.5 rat embryos. However, the effects of these growth factors are stage-specific, since similar ENS cultures established from later stage embryos (E14. 5-15.5), show markedly diminished response to GDNF and NTN. To examine whether the in vitro effects of RET activation reflect the in vivo function(s) of this receptor, the extent of programmed cell death was examined in the gut of wild-type and RET-deficient mouse embryos by TUNEL histochemistry. Our experiments show that a subpopulation of enteric NC undergoes apoptotic cell death specifically in the foregut of embryos lacking the RET receptor. We suggest that normal function of the RET RTK is required in vivo during early stages of ENS histogenesis for the survival of undifferentiated enteric NC and their derivatives.

Animals↗
PubMed1998

Regeneration of adult rat corticospinal axons induced by transplanted olfactory ensheathing cells.

Precisely localized focal stereotaxic electrolytic lesions were made in the corticospinal tract at the level of the first to second cervical segments in the adult rat. This consistently destroyed all central nervous tissue elements (axons, astrocytes, oligodendrocytes, microglia, and microvessels) in a highly circumscribed area. In a group of these rats immediately after lesioning, a suspension of cultured adult olfactory ensheathing cells was transplanted into the lesion site. Within the first week after transplantation, the cut corticospinal axons (identified by anterograde transport of biotin dextran) extended caudally along the axis of the corticospinal tract as single, fine, minimally branched sprouts that ended in a simple tip, often preceded by a small varicosity. By 3 weeks, the regenerating axons, ensheathed by P0-positive peripheral myelin had accumulated into parallel bundles, which now extended across the full length of the lesioned area and reentered the caudal part of the host corticospinal tract. The transplants contained two main types of cells: (1) p75-expressing S cells, which later formed typical peripheral one-to-one myelin sheaths around individual ensheathed axons, and (2) fibronectin-expressing A cells, which aggregated into tubular sheaths enclosing bundles of myelinated axons. The point of reentry of the axons into the central nervous territory of the caudal host corticospinal tract was marked by the resumption of oligodendrocytic myelination. Thus the effect of the transplant was to form a "patch" of peripheral-type tissue across which the cut central axons regenerated and then continued to grow along their original central pathway.

Animals↗
PubMed1998

Embryonic entorhinal transplants partially ameliorate the deficits in spatial memory in adult rats with entorhinal cortex lesions.

Our previous studies have demonstrated that axons from grafts of embryonic entorhinal cortex (EC) can reinnervate the deafferented zones in the hippocampus and form synaptic connections with the host dentate gyrus in adult mice and rats deprived of their own entorhinal inputs. Here, we have examined the ability of the EC grafts to ameliorate deficits in spatial memory. Three months after transplantation, the grafted rats and control animals were subjected to Morris water maze testing followed by histological examination. According to the exact position of grafts in the host brain, the rats with lesion and EC transplants were divided into two groups, one with EC grafts contacting both the hippocampus and overlying neocortex (n=7, EC1) and another with EC grafts confined within the hippocampus (n=6, EC2). While EC2 rats were still as impaired as those with lesion and transplants of non-entorhinal cortex (n=10, NEC) or with lesions only (n=7, LES), the EC1 rats performed better than the LES group. In a spatial memory trial, the EC1 group made more crossings over platform site and showed more focused search behavior than EC2, LES, NEC groups. The data suggest that EC grafts could partially ameliorate the deficit in spatial learning behavior in the EC-lesioned adult rats. The requirement for the graft to contact both the neocortex and the hippocampus suggests that the functional effects may be exerted by the formation of new neocortical-EC graft-hippocampal circuits.

Animals↗
PubMed1998

Transplanted embryonic entorhinal neurons make functional synapses in adult host hippocampus.

Grafts of embryonic entorhinal cortex (EC) or non-entorhinal cortex (NEC) were placed into the hippocampus of adult rats with transection of the perforant paths. Graft-host connectivity was investigated at 4-6 months post-transplantation by recording extracellular evoked responses in hippocampal slice preparations. Electrical stimulation of the grafts evoked excitatory postsynaptic potentials (EPSPs) in the outer molecular layer of the dentate gyrus, and the stratum lacunosum moleculare of CA1, CA3, and elicited population spikes in the granule cell layer and the pyramidal cell layer of CA1, but not CA3. While the latencies and the forms of these evoked response were similar to those in matched control slices from the normal animals, the amplitudes were smaller than normal controls. However, in the slices with NEC grafts, no such responses were recorded when stimulus was applied in similar position in the grafts. The findings suggest that grafted entorhinal neurons make viable synaptic connections with the host hippocampus.

Animals↗
PubMed1997

Repair of adult rat corticospinal tract by transplants of olfactory ensheathing cells.

The upper cervical corticospinal tract was transected on one side in adult rats. A suspension of ensheathing cells cultured from adult rat olfactory bulb was injected into the lesion site. This induced unbranched, elongative growth of the cut corticospinal axons. The axons grew through the transplant and continued to regenerate into the denervated caudal host tract. Rats with complete transections and no transplanted cells did not use the forepaw on the lesioned side for directed reaching. Rats in which the transplanted cells had formed a continuous bridge across the lesion exhibited directed forepaw reaching on the lesioned side.

Animals↗
PubMed1997

Endogenous synaptogenesis in the deafferented dentate gyrus does not exclude synapse formation by embryonic entorhinal transplants.

After partial deafferentation postsynaptic sites are reinnervated by local sprouting of remaining axons. We have investigated whether this process is sufficient to prevent new synapses being formed by transplanted embryonic tissue. We find that after unilateral entorhinal ablation endogenous sprouting by local axons is unable to reinnervate all the postsynaptic sites in the denervated outer dentate molecular layer. Axons from embryonic entorhinal tissue transplanted adjacent to the denervated area are able to reclaim a further proportion of the denervated postsynaptic sites. Thus, after a large lesion, endogenous sprouting is insufficient to preclude reinnervation by axons from embryonic transplants.

Afferent Pathways↗
PubMed1997

Embryonic tissue induces growth of adult axons from myelinated fiber tracts.

Suspensions of late embryonic hippocampal tissue were microinjected so as to be completely enclosed within the myelinated fiber bundles of the adult rat fimbria. Previous studies have shown that the axons from such transplanted neurons readily cross the graft/host interface and extend rapidly through the host fiber tract. The present study shows that the adult axons from the host fiber tract can also cross this interface in the opposite direction and enter the transplants. Biotin dextran tracing shows that the adult host fimbrial axons traverse the embryonic grafts and also form terminal arborizations within the transplants. Electron microscopy of orthograde electron-dense degeneration confirms that these host axons form synaptic terminals accounting for at least 6.6% of the synapses in the neuropil of the transplant. Thus, contact with embryonic nervous tissue can induce elongative growth by the adult fibers in a myelinated central tract.

Age Factors↗
PubMed1997

Integration of transplanted cultured Schwann cells into the long myelinated fiber tracts of the adult spinal cord.

A suspension of about 10,000 purified Schwann cells cultured from the neonatal rat sciatic nerve was transplanted into a discrete site in the upper cervical level of the corticospinal tract of one side in adult rats. From 4 days after transplantation immunostaining for p75 (low-affinity neurotrophin receptor) showed that the transplants consisted of a central mass of Schwann cells and cuffs of elongated Schwann cells along the perivascular space of curving blood vessels (most of which had been formed in response to the transplantation). Schwann cells leaving the central mass and perivascular cuffs migrated in strictly linear orientation along the rostrocaudal axis of the host corticospinal tract. According to the territory through which they migrated, the transplanted Schwann cells adopted two quite different forms: (1) The row Schwann cells, which migrated singly or in groups within the rows of host oligodendrocytic and astrocytic cell bodies, were non-process-bearing, rather cuboidal, brick-like cells (about 8 x 12 microm in size). (2) In contrast, the interfascicular Schwann cells, which migrated singly or intertwined in rope-like small groups interspersed among the axons of the host corticospinal tract, were larger, symmetrically bipolar cells, with processes about 100-120 microm long and 2 microm wide and bulging, ovoid nuclei, located in centrally placed cell bodies about 10 microm across. After about 6 weeks, the p75 immunoreactivity of the interfascicular Schwann cells had become down-regulated. However, from as early as 10 days after transplantation, immunostaining for the peripheral myelin protein, P0, semithin sections, and electron microscopy showed that these Schwann cells were not lost, but that they had myelinated the segments of the host corticospinal axons in the region of the transplant. In contrast, the row Schwann cells did not express P0 or form myelin. They retained their p75 immunoreactivity at long survivals (presumably because they were secluded from contacting the tract axons). The row Schwann cells also migrated farther than the interfascicular Schwann cells (possibly a function of their maintained p75 expression), becoming dispersed singly for at least 8 mm from the original transplant site. Our previous study of corticospinal tract lesions had shown the formation of a "closed" scar formed by hypertrophic astrocytic processes, which walled off a central astrocyte-free region and totally disrupted the normal longitudinal alignment of the tract astrocytic processes. In contrast, while the present Schwann cell transplants induced a comparable astrocytic hypertrophy over the same time course, the astrocytic processes remained able to penetrate the transplant site, which was not walled off, so that the longitudinal arrangement of the host corticospinal tract astrocytic skeleton was preserved intact across the region of the transplant. These observations show that Schwann cells can be intimately integrated into the cytoarchitecture of the myelinated adult host corticospinal tract. This integration is not a random dispersal in damaged areas: it involves direct interaction with the cell elements present in the host tract, it respects the complex and regular organization of the host tract glial cells, and it results in the formation of a precisely arranged mosaic of central and peripheral tissue.

Age Factors↗
PubMed1997

An urge to explain the incomprehensible: Geoffrey Harris and the discovery of the neural control of the pituitary gland.

Geoffrey Harris is responsible for our view that the brain controls the endocrine system by an exquisitely regulated pattern of synthesis and release of individual members of a family of peptide hormones. These hormones are carried through a portal vascular system that passes from the hypothalamus to the pituitary gland, where they selectively regulate the secretion of the six anterior pituitary hormones. This family of hypothalamic hormones is highly conserved in all vertebrates, including humans. They are essential for all aspects of reproduction--courtship, mating, pregnancy and young rearing--and they are responsible for the seasonal regulation of breeding. The hypothalamic control mechanism for reproduction is sexually dimorphic, with a basic female pattern that becomes masculinized under the influence of specific steroid hormones acting during development. Other members of the hypothalamic hormone family specifically regulate the secretion of pituitary growth hormone and the anterior pituitary hormones controlling the functions of the thyroid and adrenal glands. The secretion of the hypothalamic hormones is itself regulated by the feedback of the target gland hormones (such as estrogen and progesterone), which concurrently act on the brain to elicit appropriate behavior patterns. The hypothalamo-hypophysial axis plays a crucial role in the struggle for the survival of the species. By bringing the endocrine system under the control of the brain, it allows access to external environmental inputs, learned behavior patterns, and the whole of the central integrative machinery needed for the bodily functions to be sensitively and optimally adapted to the ever-changing challenges and opportunities in the outside world.

Animals↗
PubMed1997

Use of Schwann cells to induce repair of adult CNS tracts.

The ability of transplanted Schwann cells to modify the sprouts formed by cut central axons, and in particular to induce branching and extension of axon sprouts, is an encouraging sign for their possible future use in repair. The accessibility of the Schwann cells in the culture stage before transplantation offers a practical opportunity for genetic engineering (e.g. to introduce genes directing the expression of specific growth factors) which might be useful in designing a future method for the repair of human spinal injury. It must be borne in mind, however, that even the most successful cases of peripheral nerve grafts have shown only a limited proportion of axons growing back from the grafts into the environment of the CNS (Carter et al., 1989). When we constructed Schwann cells transplanted into the thalamus (Brook et al., 1994), we did not observe axons leaving the artificial tracts. In our experiments with Schwann cells transplanted into the spinal cord (Li & Raisman, 1994), the axons have only been studied within the graft, and we have as yet not been able to assess the extent to which they re-enter the CNS. For effective regeneration to occur, regenerating axons must not only be able to re-enter their original pathways and elongate along them, but also leave them in a correct manner--i.e. by making appropriate choices from a wide range of destinations. Therefore the effectiveness of a Schwann cell "bridging" repair must depend upon the self-organising capacity of the adult CNS (e.g. Florence et al., 1996).

Adult↗
PubMed1996

Conditionally immortalized neural progenitor cell lines integrate and differentiate after grafting to the adult rat striatum. A combined autoradiographic and electron microscopic study.

Neural progenitor cell lines, generated by conditional immortalization from the embryonic CNS, have previously been shown to survive and integrate after transplantation to the adult brain. The present study was designed to investigate the in vivo differentiation and morphological features of grafted neural progenitors using combined autoradiography and transmission electron microscopy of two temperature-sensitive neural progenitor cell lines, HiB5 and ST14A, labeled with 3H-thymidine prior to grafting. Two weeks after transplantation to the striatum the cells were found dispersed over an area extending about 1.5 mm from the injection site. Labeled cells located within the myelinated fiber bundles of the internal capsule were closely associated with myelinated axons and presented profiles similar to oligodendrocytes, while most of the grafted cells in the grey matter had morphological features of astroglia. Some labeled cells occurred also in close association with small blood vessels, morphologically resembling host pericytes. The results show that the immortalized neural progenitors can differentiate into mature glial cells, including astrocytes, oligodendrocytes and pericytes, after implantation into the adult striatum. The ability of the cells to become fully integrated with the resident glial population suggests that they will be highly useful as vehicles for intracerebral transgene expression in ex vivo gene transfer.

Animals↗