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Biomedical subjects

L S Shihabuddin

Publications and source records attributed to L S Shihabuddin.

10 recordsLinked to original sources
PubMed2004

Intracerebral transplantation of adult mouse neural progenitor cells into the Niemann-Pick-A mouse leads to a marked decrease in lysosomal storage pathology.

Niemann-Pick disease is caused by a genetic deficiency in acid sphingomyelinase (ASM) leading to the intracellular accumulation of sphingomyelin and cholesterol in lysosomes. In the present study, we evaluated the effects of direct intracerebral transplantation of neural progenitor cells (NPCs) on the brain storage pathology in the ASM knock-out (ASMKO) mouse model of Type A Niemann-Pick disease. NPCs derived from adult mouse brain were genetically modified to express human ASM (hASM) and were transplanted into multiple regions of the ASMKO mouse brain. Transplanted NPCs survived, migrated, and showed region-specific differentiation in the host brain up to 10 weeks after transplantation (the longest time point examined). In vitro, gene-modified NPCs expressed up to 10 times more and released five times more ASM activity into the culture media compared with nontransduced NPCs. In vivo, transplanted cells expressed hASM at levels that were barely detectable by immunostaining but were sufficient for uptake and cross-correction of host cells, leading to reversal of distended lysosomal pathology and regional clearance of sphingomyelin and cholesterol storage. Within the host brain, the area of correction closely overlapped with the distribution of the hASM-modified NPCs. No correction of pathology occurred in brain regions that received transplants of nontransduced NPCs. These results indicate that the presence of transduced NPCs releasing low levels of hASM within the ASMKO mouse brain is necessary and sufficient to reverse lysosomal storage pathology. Potentially, NPCs may serve as a useful gene transfer vehicle for the treatment of CNS pathology in other lysosomal storage diseases and neurodegenerative disorders.

Animals↗
PubMed2000

Adult spinal cord stem cells generate neurons after transplantation in the adult dentate gyrus.

The adult rat spinal cord contains cells that can proliferate and differentiate into astrocytes and oligodendroglia in situ. Using clonal and subclonal analyses we demonstrate that, in contrast to progenitors isolated from the adult mouse spinal cord with a combination of growth factors, progenitors isolated from the adult rat spinal cord using basic fibroblast growth factor alone display stem cell properties as defined by their multipotentiality and self-renewal. Clonal cultures derived from single founder cells generate neurons, astrocytes, and oligodendrocytes, confirming the multipotent nature of the parent cell. Subcloning analysis showed that after serial passaging, recloning, and expansion, these cells retained multipotentiality, indicating that they are self-renewing. Transplantation of an in vitro-expanded clonal population of cells into the adult rat spinal cord resulted in their differentiation into glial cells only. However, after heterotopic transplantation into the hippocampus, transplanted cells that integrated in the granular cell layer differentiated into cells characteristic of this region, whereas engraftment into other hippocampal regions resulted in the differentiation of cells with astroglial and oligodendroglial phenotypes. The data indicate that clonally expanded, multipotent adult progenitor cells from a non-neurogenic region are not lineage-restricted to their developmental origin but can generate region-specific neurons in vivo when exposed to the appropriate environmental cues.

Animals↗
PubMed1999

Truncated BRCA2 is cytoplasmic: implications for cancer-linked mutations.

BRCA2 mutations predispose carriers mainly to breast cancer. The vast majority of BRCA2 mutations are predicted to result in a truncated protein product. The smallest known cancer-associated deletion removes from the C terminus only 224 of the 3,418 residues constituting BRCA2, suggesting that these terminal amino acids are crucial for BRCA2 function. A series of green fluorescent protein (GFP)-tagged BRCA2 deletion mutants revealed that nuclear localization depends on two nuclear localization signals that reside within the final 156 residues of BRCA2. Consistent with this observation, an endogenous truncated BRCA2 mutant (6174delT) was found to be cytoplasmic. Together, these studies provide a simple explanation for why the vast majority of BRCA2 mutants are nonfunctional: they do not translocate into the nucleus.

BRCA2 Protein↗
PubMed1999

The search for neural progenitor cells: prospects for the therapy of neurodegenerative disease.

The etiology of many neurodegenerative diseases has been identified in recent years. Treatment of central nervous system (CNS) disease could focus on one or more steps that lead to cell loss. In the past decade, cell therapy and/or ex vivo gene therapy have emerged as possible strategies for the treatment of neurodegenerative diseases. The ability to grow CNS-derived neural progenitor cells using growth factors has been extremely useful to study diverse phenomena including lineage choice, commitment and differentiation. By virtue of their biological properties and their presence in the adult CNS, neural progenitors represent good candidates for multiple cell-based therapies for neural diseases. Further identification of the molecules that direct the differentiation of adult neural progenitors may allow their activation in vivo to induce self-repair. This review addresses the nature, distribution and regulation of neural stem cells and the potential for applying these cells to both structural CNS repair and gene therapy.

Adult↗
PubMed1997

FGF-2 is sufficient to isolate progenitors found in the adult mammalian spinal cord.

The adult rat brain contains progenitor cells that can be induced to proliferate in vitro in response to FGF-2. In the present study we explored whether similar progenitor cells can be cultured from different levels (cervical, thoracic, lumbar, and sacral) of adult rat spinal cord and whether they give rise to neurons and glia as well as spinal cord-specific neurons (e.g., motoneurons). Cervical, thoracic, lumbar, and sacral areas of adult rat spinal cord (>3 months old) were microdissected and neural progenitors were isolated and cultured in serum-free medium containing FGF-2 (20 ng/ml) through multiple passages. Although all areas generated rapidly proliferating cells, the cultures were heterogeneous in nature and cell morphology varied within a given area as well as between areas. A percentage of cells from all areas of the spinal cord differentiate into cells displaying antigenic properties of neuronal, astroglial, and oligodendroglial lineages; however, the majority of cells from all regions expressed the immature proliferating progenitor marker vimentin. In established multipassage cultures, a few large, neuron-like cells expressed immunoreactivity for p75NGFr and did not express GFAP. These cells may be motoneurons. These results demonstrate that FGF-2 is mitogenic for progenitor cells from adult rat spinal cord that have the potential to give rise to glia and neurons including motoneurons.

Animals↗
PubMed1996

Selective hippocampal lesions differentially affect the phenotypic fate of transplanted neuronal precursor cells.

RN33B cells, a CNS-derived neuronal precursor cell line, transplanted into normal adult rat hippocampus can survive and morphologically differentiate with their ultimate morphology dependent on the integration site. This study examined the differentiation and structural integration of RN33B cells transplanted into the lesioned adult hippocampus. Pyramidal neurons of the CA1-3 regions or granular neurons in the dentate gyrus were preferentially destroyed by unilateral intraventricular kainic-acid or intradentate colchicine injections, respectively. One week after the lesion, a suspension of undifferentiated beta-galactosidase (beta-gal)-labeled RN33B cells was stereotaxically transplanted into the lesioned or the contralateral hippocampus. After 5-7 weeks, sections of the recipient brains were analyzed by toluidine blue staining and immunohistochemistry for beta-gal, GFAP, and OX-42. A reactive gliosis was observed on the lesioned side which persisted up to 7 weeks postlesion (the latest time point examined). RN33b cells survived in the lesioned hippocampus and assumed variable morphologies depending on the hippocampal layer into which they integrated. Only RN33B cells located in intact or partially damaged cell layers or in the unlesioned contralateral hippocampus differentiated with morphologies similar to those of endogenous neurons characteristic of those layers. Cells located in layers completely depleted of endogenous neurons assumed bipolar morphologies or sent out multiple processes with no structural polarity, unlike the neuronal morphologies characteristically seen in intact hippocampal cell layers. These data suggest that the presence of some endogenous neurons and partially conserved cytoarchitectural organization are essential for immortalized neuroepithelial precursor cells to differentiate into region-specific neuronal cell types.

Animals↗
PubMed1996

Induction of mature neuronal properties in immortalized neuronal precursor cells following grafting into the neonatal CNS.

RN33B, a conditionally-immortalized neuronal cell line, survives and differentiates following grafting into the neocortex and hippocampus of adult and neonatal rat hosts. We have previously shown that these cells assume shapes characteristic of endogenous neurons at the integration site and persist up to 24 weeks post-grafting. In the present study we use electron microscopy and immunohistochemistry to characterize such cells. Differentiated RN33B cells were identical in size to endogenous neurons and their sizes depended on the specific location of integration. RN33B cells in the granule cell layer of the dentate gyrus and CA3 and CA1 pyramidal layers were 9.0, 15.3, and 12.6 microns in diameter, respectively. Grafted RN33B cells received synapses from fibres of host origin. Differentiated cells expressed neuronal markers, but not glial markers. Some differentiated cells expressed glutamate both in vitro and in vivo whereas undifferentiated cells did not. Grafted RN33B cells that differentiated with morphologies similar to CA3 pyramidal neurons and pyramidal cortical neurons expressed Py antigen, a neuronal marker that is differentially expressed in endogenous large pyramidal neurons of the cerebral cortex and large pyramids of hippocampal field CA3. This Py immunoreactivity was region-specific and corresponded to the endogenous pattern of Py immunostaining. Collectively, these data indicate that RN33B cells are capable of region-specific differentiation and have the potential to integrate functionally into the host CNS.

Animals↗
PubMed1995

The adult CNS retains the potential to direct region-specific differentiation of a transplanted neuronal precursor cell line.

The chronic survival and differentiation of the conditionally immortalized neuronal cell line, RN33B, was examined following transplantation into the adult and neonatal rat hippocampus and cerebral cortex. In clonal culture, differentiated RN33B cells express p75NTR and trkB mRNA and protein, and respond to brain-derived neurotrophic factor treatment by inducing c-fos mRNA. Transplanted cells, identified using immunohistochemistry to detect beta-galactosidase expression, were seen in most animals up to 24 weeks posttransplantation (the latest time point examined). Stably integrated cells with various morphologies consistent with their transplantation site were observed. In the cerebral cortex, many RN33B cells differentiated with morphologies similar to pyramidal neurons and stellate cells. In the hippocampal formation, many RN33B cells assumed morphologies similar to pyramidal neurons characteristic of CA1 and CA3 regions, granular cell layer neurons of the dentate gyrus, and polymorphic neurons of the hilar region. Identical morphologies were observed in both adult and neonatal hosts, although a greater percentage of beta-galactosidase immunoreactive cells had differentiated in the neonatal brains. These results suggest that RN33B cells have the developmental plasticity to respond to local microenvironmental signals and that the adult brain retains the capacity to direct the differentiation of neuronal precursor cells in a direction that is consistent with that of endogenous neurons.

Aging↗
PubMed1993

The role of previous nociceptive input in development of autotomy following cordotomy.

Intractable pains have been described after surgical or accidental lesions in the peripheral or central nervous system. The possible contribution of antecedent injury to the appearance of these pains was examined in an animal model for chronic pain which involved observing self-mutilation or autotomy behavior in rats. Various combinations of previous injury, selective spinal cut, and peripheral denervation were carried out on rats. Injuries ranged from mild to moderate and were produced by nociceptive stimuli from formaldehyde injection, induced local arthritis, or hot water application. Selective spinal cuts included either a dorsal column (DC), a dorsal quadrant which included a DC and a dorsolateral funiculus, an anterolateral column (ALC), or a hemisection. In rats without prior exposure to injury the various types of spinal cuts were not associated with any autotomy. In rats with prior exposure to formaldehyde injection, autotomy was associated only with spinal cuts that involved the ALC. In an otherwise similar group of rats but with prior induction of local arthritis, a stronger association of ALC lesion and autotomy was observed. In an earlier study, rats with ALC lesion prior to denervation showed reduced autotomy. In this study, we demonstrated that in rats with previous exposure to heat injury, an ALC lesion was strongly associated with autotomy. Autotomy was absent, however, in rats with heat injury only. These findings strongly suggest that pain resulting from previous exposure to injury produces a memory trace in the central nervous system which can account for the phantom pains encountered in various clinical conditions.

Animals↗