Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.4.37 (CNPase)
539 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have established a permanent cell line (1H91) of putative type-1 astrocyte precursor cells that were clonally derived from a single cell isolated from E16 mouse cerebellum. Epidermal growth factor (EGF) and transforming growth factor (TGF alpha) are strong mitogens for 1H91 cells (ED50 of 9.02 + 1.74 ng/ml and 15.98 +/- 2.34 ng/ml, respectively), while basic fibroblast growth factor (bFGF) is only weakly mitogenic and platelet derived growth factor (PDGF) has no mitogenic activity. In the proliferative state, the 1H91 cells are immunohistochemically positive for nestin and vimentin, and negative for A2B5, CNPase, neurofilament (NF), and neuron specific enolase (NSE). The majority of EGF-treated 1H91 cells are not immunoreactive for glial acid fibrillary protein (GFAP). In the presence of 5 ng/ml bFGF, 1H91 cells become non-mitotic and develop a morphology consistent with a fibrous astrocyte. In contrast to the proliferating cultures, the bFGF treated cultures were strongly immunoreactive for GFAP, only mildly immunoreactive for nestin and vimentin, and negative for A2B5, CNPase, NF, and NSE. Type-1 astrocytes are known to proliferate in response to EGF, and are immunohistochemically GFAP positive, A2B5 negative, and CNPase negative [38]. However, type-1 astrocytes only develop a fibrous morphology during the process of reactive gliosis [31]. Since EGF is a strong mitogen for 1H91 cells, and these cells may be differentiated into GFAP positive, A2B5 negative, CNPase negative astrocytes, we conclude that 1H91 cells conform to a type-1 astrocyte precursor phenotype. In addition, the fibrous morphology of the bFGF treated 1H91 cells suggests that these cells follow the process of reactive gliosis. Therefore, the 1H91 clonal cell line may provide an in vitro model for studying the underlying cellular mechanisms of the type-1 astrocyte in reactive gliosis.
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PMID:Isolation, cloning and characterization of a putative type-1 astrocyte cell line. 912 27

MHP36 is a nestin bFGF-dependent cell line isolated from embryonic hippocampus using a thermolabile form of SV40 T antigen. When grafted in ischemic hippocampus MHP36 cells differentiate and alleviate the cognitive deficit associated with the lesion. We report here in vitro features of MHP36 cells. First, we found that T Ag expression was not necessary for MHP36 growth as cells cultured at the nonpermissive temperature carry on proliferating at a normal rate, Second, we observed that part of MHP36 cells spontaneously differentiate into astrocytes when bFGF is removed at39 degrees C. This differentiation was increased 4-fold by leukemia inhibitory factor. Third, we found that the majority of cells spontaneously expressed oligodendrocytic markers (CNPase, A2B5, GalC) when cultured at low density.
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PMID:Regulation of glial differentiation of MHP36 neural multipotent cell line. 1144 41

Lineage uncommitted pluripotent stem cells reside in the connective tissue of skeletal muscle. The present study was carried out with pluripotent stem cells (PPSCs) isolated from 6-month old rat muscle. Before differentiation, these cells were vimentin+, CD90+, CD45-, and varied in their expression of CD34. The PPSCs were expanded as non-adherent aggregates under similar conditions to those used to generate neurospheres from embryonic or neural stem cells. The PPSC-derived neurospheres were positive for nestin, an early marker present in neuronal precursors, and expressed the two alternative mRNA forms of the neuroectodermal marker Pax-6, as well as mRNA for Oct-4, a gene related to the pluripotentiality of stem cells. To confirm their neural potential, PPSC-derived neurospheres were plated on coated coverslips under varying conditions: Neurobasal medium with N2 or B27, and either NT3 or BDNF. After 4-6 days the cells expressed neuronal (Tuj1+, NF68), astrocytic (GFAP) and oligodendrocytic (MOSP+, MBP+) markers, both by immunocytochemistry and RT-PCR. In addition, PPSCs were cultured as monolayers under adherent conditions, exposed to growth factors and defined differentiating conditions for 5 hr, and subsequently kept for 2 days in a maturation medium. At this point they gave rise to a mixed population of early neural progenitors (Nestin+ or NG2+), immature and mature neurons (Tuj1+ and NF145+) and myelin producing oligodendrocytes (CNPase + and MOSP+). Our study shows that PPSCs present in adult muscle can overcome germ lineage restrictions and express the molecular characteristics of brain cells. Therefore, PPSCs isolated from adult muscle could provide a novel source for autologous cell replacement in neurodegenerative and demyelinating diseases.
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PMID:Neuronal differentiation of stem cells isolated from adult muscle. 1220 82

Progenitors that migrate through the white matter of the postnatal cerebellum give rise to cortical interneurons, astroglia, and oligodendroglia. To determine whether this progenitor population is heterogeneous with respect to specific lineage markers, we infected progenitors in vivo with a retrovirus encoding the green fluorescent protein on postnatal day 4/5 and labeled them in situ with various antibodies 2 days postviral injection: the neuronal marker was the transcription factor SOX1; early oligodendroglial markers were chondroitin sulfate proteoglycan antigen and platelet-derived growth factor receptor-alpha. Markers for astroglial progenitors were vimentin, nestin, zebrin II, and the astroglial-specific glutamate transporter subtype GLAST. None of the progenitors was doubly labeled with any combination of markers characteristic for different cell lineages. Most progenitors were not labeled with any of the various combinations of antibodies used. Progenitors did not express markers characteristic for mature astroglia (GFAP), oligodendroglia (CNPase), or neurons (MAP2). Thus, although these progenitors are morphologically indistinguishable, a minority expresses markers of early neuronal or glial lineages, suggesting that they begin to differentiate during migration.
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PMID:Progenitors in the postnatal cerebellar white matter are antigenically heterogeneous. 1227 92

Neural stem/progenitor cells are clonogenic in vitro and produce neurospheres in serum-free medium containing epidermal growth factor (EGF) and fibroblast growth factor (FGF2). Here, we demonstrate that lysophosphatidic acid (LPA) instigated the clonal generation of neurospheres from dissociated mouse postnatal forebrain in the absence of EGF and FGF2. LPA induced proliferation of cells which co-expressed Sca-1 antigen and AC133, markers of primitive hematopoietic and neural stem/progenitor cells. Clonal expansion of these cells induced by LPA was inhibited by diacylglycerol- pyrophosphate (DGPP), an antagonist of the LPA receptor subtypes LPA1 and LPA3. Moreover, Sca-1- and AC133-positive cells of these neurospheres expressed LPA1, LPA2, and LPA3, suggesting important roles for these LPA receptors in proliferation of neural progenitors. LPA induced neurospheres to differentiate on an adherent laminin/poly-L-ornithine matrix. In differentiating neurospheres, LPA receptors co-localized with betaIII-tubulin, nestin, and CNPase, but not with glial fibrillary acidic protein (GFAP), a marker of astrocyte lineage. Our results demonstrate for the first time that lysophosphatidic acid induces clonal neurosphere development via proliferation of AC133/Sca-1-positive stem cells by a receptor-dependent mechanism. This differentiation was characterized by the initial co-localization of neural specific antigens at sites of LPA receptor expression upon their interaction with the inducing agonist.
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PMID:Lysophosphatidic acid induces clonal generation of mouse neurospheres via proliferation of Sca-1- and AC133-positive neural progenitors. 1568 36

This study aims to investigate the therapeutic potential of adult bone marrow stromal cells (BMSCs). Exposed to a cocktail of induction medium, some BMSCs could differentiate into cell types with phenotypes of neural lineages in vitro. These cells expressed neural markers nestin, GFAP, 68-kDa neurofilament and beta-tubulin III as detected by immunohistochemistry and RT-PCR. Fluorescence-labeled cells were injected intravenously at 72 h after traumatic brain injury. Transplanted cells survived and migrated to the ipsilateral cerebral cortex at different time points after injection. They were immunopositive for neuronal marker MAP-2, oligodendrocyte marker CNPase, astrocytic maker GFAP or microglial marker OX-42 in vivo. In rats receiving BMSC transplants, there were significant improvements in motor and neurological functions when compared with the control groups. Hence, the therapeutic potential of BMSCs for traumatic brain injury is further amplified.
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PMID:Adult bone marrow cells differentiate into neural phenotypes and improve functional recovery in rats following traumatic brain injury. 1645 99

Several lines of evidence have suggested roles for pituitary adenylate cyclase-activating polypeptide (PACAP) in the developing nervous system. Previously, we showed that mRNA for PACAP, vasoactive intestinal peptide (VIP), and their three receptor subtypes, is differentially expressed in embryonic stem (ES) cells, ES cell-derived, neural stem cell-enriched cultures, and differentiated neurons, by using the five steps of the in vitro neuronal culture model of ES cell differentiation. Here, we examined the effects of PACAP on self-renewal and cell lineage determination of neural progenitor/stem cells. PACAP inhibited the basic fibroblast growth factor-induced proliferation (self-renewal), as assessed by neurosphere formation. PACAP increased microtubule-associated protein 2-positive neurons without affecting the number of cells positive for the neural stem cell marker nestin, astrocyte marker glial fibrillary acidic protein, and oligodendrocyte marker CNPase. These results suggest that PACAP inhibits self-renewal but, instead, induces early neuronal differentiation of neural progenitor cells.
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PMID:Inhibition of self-renewal and induction of neural differentiation by PACAP in neural progenitor cells. 1688 89

Transplantation of neural stem cells (NSC) into lesioned spinal cord offers the potential to increase regeneration by replacing lost neurons or oligodendrocytes. The majority of transplanted NSC, however, typically differentiate into astrocytes that may exacerbate glial scar formation. Here we show that blocking of ciliary neurotrophic factor (CNTF) with anti-CNTF antibodies after NSC transplant into spinal cord injury (SCI) resulted in a reduction of glial scar formation by 8 weeks. Treated animals had a wider distribution of transplanted NSC compared with the control animals. The NSC around the lesion coexpressed either nestin or markers for neurons, oligodendrocytes, or astrocytes. Approximately 20% fewer glial fibrillary acidic protein-positive/bromodeoxyuridine (BrdU)-positive cells were seen at 2, 4, and 8 weeks postgrafting, compared with the control animals. Furthermore, more CNPase(+)/BrdU(+) cells were detected in the treated group at 4 and 8 weeks. These CNPase(+) or Rip(+) mature oligodendrocytes were seen in close proximity to host corticospinal tract (CST) and 5HT(+) serotonergic axon. We also demonstrate that the number of regenerated CST fibers both at the lesion and at caudal sites in treated animals was significantly greater than that in the control animals at 8 weeks. We suggest that the blocking of CNTF at the beginning of SCI provides a more favorable environment for the differentiation of transplanted NSC and the regeneration of host axons.
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PMID:Neutralization of ciliary neurotrophic factor reduces astrocyte production from transplanted neural stem cells and promotes regeneration of corticospinal tract fibers in spinal cord injury. 1704 31

The superoxide dismutase 1(G93A G1H) (SOD1(G93A G1H)) transgenic mouse is a model of familial human amyotrophic lateral sclerosis (ALS) that has progressive neurodegeneration within the spinal cord and brainstem. In this study, we investigated the number and differentiation of neural precursor cells (NPCs). Nestin-positive NPCs were rarely seen in the nervous system of wild type controls or pre-disease mice at post-natal days 30 and 60. With disease onset on post-natal day 90, nestin labeled NPCs proliferated preferentially in the brainstem with maximal number and density at post-natal day 120. NPCs did not double-label with CNPase or O(4) markers of oligodendrocytes. The majority of the NPCs co-labeled with the astrocyte maker glial fibrillary acidic protein (GFAP) and a small number with the neuronal marker NeuN. At disease onset, 73 and 10% of NPCs co-expressed GFAP and NeuN respectively while at severe disease stage, 80 and 8% of the NPCs co-expressed GFAP and NeuN. Proliferating cell nuclear antigen (PCNA) was used to confirm that at least some of these cells undergo mitosis. Future studies could be directed at controlling the differentiation of these endogenous NPCs into neurons and astrocytes in order to ameliorate the degeneration within the brainstem of the ALS mouse.
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PMID:Increased number and differentiation of neural precursor cells in the brainstem of superoxide dismutase 1(G93A) (G1H) transgenic mouse model of amyotrophic lateral sclerosis. 1743 5

Adult multipotent stem cells have been isolated from various non-neural tissues. Here, we report the isolation of multipotent stem cells from rat periodontal ligament (PDL) using neurosphere-forming culture system. Enzymatically dissociated PDL cells were cultured in serum-free basal medium containing EGF, bFGF, and LIF. Free-floating spheres expressing nestin, GFAP, and vimentin were formed by 7 days of the culture. In addition, spheres expressed mRNA of neural crest-associated transcription factors Twist, Slug, Sox2, and Sox9. PDL-derived spheres differentiated into multinucleated myotubes, NFM-positive neuron-like cells, GFAP-positive astrocyte-like cells and CNPase-positive oligodendrocyte-like cells. Methylcellulose colony-forming assay revealed that a single PDL cell could form a sphere at a frequency of approximately 0.01% of total cells. These data indicate that PDL-derived spheres contained multipotent adult stem cells capable of differentiating into both neural and mesodermal progeny. This is the first report of the isolation of PDL-derived stem cells with primitive neural crest stem cell features.
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PMID:Isolation of multipotent stem cells from adult rat periodontal ligament by neurosphere-forming culture system. 1745 43


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