Use of a species-specific antibody for demonstrating mouse neurons transplanted to rat brains

Lund, Raymond D.; Chang, Fen‐Lei; Hankin, Mark; Lagenaur, Carl F.

doi:10.1016/0304-3940(85)90428-8

Cited by 82 publications

(37 citation statements)

References 8 publications

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“…2H, yellow coexpression; ref. 30) and could be identified also by mouse-specific intranuclear fluorescent bodies after labeling with bisbenzimide (Hoechst; ref. 31; Fig.…”

Section: Es Cells Differentiate Into Mesencephalic Dopaminergic-like mentioning

confidence: 99%

Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model

Björklund

Sánchez‐Pernaute

Chung

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

1,087

670

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Although implantation of fetal dopamine (DA) neurons can reduce parkinsonism in patients, current methods are rudimentary, and a reliable donor cell source is lacking. We show that transplanting low doses of undifferentiated mouse embryonic stem (ES) cells into the rat striatum results in a proliferation of ES cells into fully differentiated DA neurons. ES cell-derived DA neurons caused gradual and sustained behavioral restoration of DA-mediated motor asymmetry. Behavioral recovery paralleled in vivo positron emission tomography and functional magnetic resonance imaging data demonstrating DA-mediated hemodynamic changes in the striatum and associated brain circuitry. These results demonstrate that transplanted ES cells can develop spontaneously into DA neurons. Such DA neurons can restore cerebral function and behavior in an animal model of Parkinson's disease. P arkinson's disease (PD) is a degenerative disorder characterized by a loss of midbrain dopamine (DA) neurons with a subsequent reduction in striatal DA (1). Pharmacological treatment with L-DOPA works initially, but reduced efficacy and development of motor complications requires treatment alternatives such as deep brain stimulation and fetal DA neuron transplantation (2). There is evidence both from animal models and clinical investigations showing that fetal DA neurons can produce symptomatic relief (3-6). Technical and ethical difficulties in obtaining sufficient and appropriate donor fetal brain tissue have limited the application of this new therapy.Previous work showed that DA neurons can be produced in vitro from ventral mesencephalic (VM) precursor cells (7). A problem using expanded fetal VM precursors (7) is the low in vivo survival rate of 3-5% of the grafted DA neurons, which eliminates the actual gain by in vitro cell number expansion compared with fresh (unexpanded) fetal day-12 VM (7,8).Embryonic stem (ES) cells have many characteristics required for an optimal cell source for cell-replacement therapy (9, 10). ES cells are self-renewing and multipotent cells derived from the inner cell mass of the preimplantation blastocyst (11). We have shown previously that mouse ES cells transplanted to normal mice or 6-hydroxydopamine (OHDA)-lesioned rats can differentiate spontaneously into tyrosine hydroxylase (TH)-positive and serotonin (5HT)-positive neurons. This differentiation is likely not caused by a specific inductive signal from the host brain, because similar neuronal differentiation occurs after placement in the kidney capsule (11). In our previous study (11), grafts frequently showed heterogeneous morphology and often became very large, disrupting the cytoarchitecture at the implantation site, which prevented the possibility for functional integration. Because neurons develop from ES cells even when implanted outside the central nervous system (11) and ectoderm develops into neural tissue when cellto-cell communication is disrupted by dissociation of the cells (12-14), we hypothesized that dilution of ES cells into single-cell suspension...

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“…2H, yellow coexpression; ref. 30) and could be identified also by mouse-specific intranuclear fluorescent bodies after labeling with bisbenzimide (Hoechst; ref. 31; Fig.…”

Section: Es Cells Differentiate Into Mesencephalic Dopaminergic-like mentioning

confidence: 99%

Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model

Björklund

Sánchez‐Pernaute

Chung

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

1,087

670

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“…In order to label entire cells and their processes, we used an additional antibody, M6. This antibody recognizes an antigen that is present on the surface of mouse neurons and their processes (Lund et al, 1985). …”

Section: Dendrite Identificationmentioning

confidence: 99%

“…In order to identify clearly the full extent of the processes from cortical neurons, embryonic (E 18) mouse cortical neurons were cocultured onto rat astroglial monolayers and the antibody M6 was used to label selectively the mouse neurons (Lund et al, 1985). The M6 antigen was distributed on all processes of the neurons, with patches of continuous labeling alternating with punctate staining.…”

Section: Glial Characterizationmentioning

confidence: 99%

Regional differences in glial-derived factors that promote dendritic outgrowth from mouse cortical neurons in vitro

Roux

Reh

1994

J. Neurosci.

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To determine whether glia from different CNS regions differ in their ability to support axons or dendrites, embryonic (E18) mouse cortical neurons were cocultured with early postnatal (P4) rat astroglial derived from cortex, retina, olfactory bulb, mesencephalon, striatum, and spinal cord. After 5 d in vitro, axon and dendrite outgrowth from isolated neurons was quantified with double-labeling immunohistochemical techniques. Whereas axonal growth was similar on the various monolayers, total primary dendritic outgrowth was nearly threefold greater on glia derived from the cortex, retina, and olfactory bulb than on glia derived from mesencephalon, striatum, or spinal cord. This effect was principally on the number of primary dendrites rather than the elongation of individual dendrites. Similar morphological differences were observed when cortical neurons were grown on polylysine in a noncontact coculture system with glia continuously conditioning the media. This selective promotion of dendrite growth was independent of neuron survival. These results indicate that there are regional differences in the ability of CNS glia to support dendritic growth and that this effect is due, in part, to release of a diffusable factor.

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“…In the majority of experiments, we relied on interspecies cocultures, placing mouse explants on rat cells and visualizing the mouse explant neurites selectively with the M6 monoclonal antibody (Lund et al, 1985), specific to mouse neurons.…”

Section: Methodsmentioning

confidence: 99%

“…To identify murine pontine fibers, the cultures were immunostained with the mouse-specific M6 monoclonal antibody, generously provided by Dr. Carl Lagenauer (Lund et al, 1985). Neurites were identified by immunostaining with the NF2 monoclonal antibody against the 200,000 Da neurofilament protein (Liem et al, 1985), generously provided by our colleague Dr. R. K. H. Liem.…”

Section: Methodsmentioning

confidence: 99%

Cerebellar target neurons provide a stop signal for afferent neurite extension in vitro

1992

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The contributions of cell-cell interactions to the establishment of specific patterns of innervation within target brain regions are not known. To provide an experimental analysis of the regulation of afferent axonal growth, we have developed an in vitro assay system, based on the developing mouse cerebellum, in which afferent axons from a brainstem source of mossy fiber afferents, the basilar pontine nuclei, were cocultured with astroglia or granule neurons purified from the cerebellum. In the absence of cells from the cerebellum, pontine explants produced axons that fasciculated and extended rapidly on a culture surface treated with poly-lysine or laminin. When pontine neurites grew onto cerebellar astroglial cells, outgrowth was more abundant than on substrates alone, suggesting that glial cells provide a positive signal for axon extension. Time-lapse video microscopy indicated that the rate of neurite extension increased from less than 50 microns/hr to more than 100 microns/hr when axonal growth cones moved from the culture substratum onto an astroglial-cell surface. Acceleration of neurite extension was also observed as pontine neurites grew onto other pontine neurites. By contrast, when pontine neurites grew on granule neurons, the appropriate targets of mossy fibers, the length of pontine neurites was greatly reduced. As growing axons terminated on granule neurons, the target cells appeared to provide a “stop-growing signal” for axon extension. The length of pontine neurites decreased with increasing granule neuron density. Two lines of evidence suggested that the stop signal was contact mediated. First, video microscopy showed that pontine growth cones stopped extending after contacting a granule neuron. Second, the length of afferent axons was not reduced when pontine neurites grew at a distance from granule neurons. Competition experiments where both astroglia and granule neurons were plated together suggested that the growth arrest signal provided by granule neurons could override the growth-promoting signal provided by astroglial cells. These results suggest that specific cell- cell interactions regulate the growth of pontine afferent axons within their cerebellar target, with axoaxonal and axoglial interactions promoting axon extension and axon-target cell interactions interrupting axon extension.

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Use of a species-specific antibody for demonstrating mouse neurons transplanted to rat brains

Cited by 82 publications

References 8 publications

Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model

Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model

Regional differences in glial-derived factors that promote dendritic outgrowth from mouse cortical neurons in vitro

Cerebellar target neurons provide a stop signal for afferent neurite extension in vitro

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