Biomedical subjects

D Pleasure

Publications and source records attributed to D Pleasure.

At least 37 records · Page 2Linked to original sources

PubMed1998

Schwann cell undergoes apoptosis during experimental allergic neuritis (EAN).

Schwann cell apoptosis is not detectable in the normal mature mammalian peripheral nervous system (PNS). However, during PNS cell-mediated demyelination, apoptosis contributes to the elimination of endoneurial T-lymphocytes. We report here that approximately 10% of Schwann cells die by apoptosis during the early phases of recovery from experimental autoimmune neuritis (EAN) in the adult rat, a model for the Guillain-Barrè syndrome. Schwann cell apoptosis, follows endoneurial T-cell clearance, and is prominent in the nerve roots, the site of most severe segmental demyelination, but is rare in the more distal regions of the PNS, where Wallerian degeneration predominates. Further immunological analysis showed that the p75 neurotrophin receptor (p75NTR) is expressed in 2% of both apoptotic and non apoptotic Schwann cells, while Ki-67, a marker of cell proliferation, is expressed in 20% of apoptotic and in 1% of non apoptotic Schwann cells. Our new observations indicate that apoptosis during cell-mediated demyelination can be a phenomenon related either to the development or the recovery of autoimmune cell mediated inflammation.

Animals↗

PubMed1998

Nerve growth factor maintains regulation of intracellular calcium in neonatal sympathetic neurons but not in mature or aged neurons.

We examined the effects of nerve growth factor on the regulation of intracellular calcium levels of superior cervical ganglion neurons in terms of postnatal maturation and ageing. Rat superior cervical ganglion neurons from three age groups (neonatal: 0 to one-day-old, young adult: three to six-month-old, and aged: more than 24-month-old) were dissociated and cultured in the presence or absence of 100 ng/ml of nerve growth factor. Intracellular free calcium levels ([Ca2+]i) were measured using the fura-2 microfluorometry. Nerve growth factor treatment increased the resting [Ca2+]i of neonatal neurons, although it had no effect on those of mature and aged neurons. We further examined the effects of nerve growth factor on the transient increase of [Ca2+]i induced by methacholine (0.1 mM), caffeine (20 mM) or high-potassium medium (40 mM K+). Nerve growth factor pre-treatment significantly increased the population of neonatal superior cervical ganglion neurons which responded to methacholine, whereas almost all young adult and aged neurons responded to methacholine regardless of pre-treatment of nerve growth factor. Caffeine induced a cyclic alteration of [Ca2+]i (oscillation) in 45% of the neonatal superior cervical ganglion neurons when they were maintained without nerve growth factor, but nerve growth factor treatment suppressed the oscillation to 10% of neurons. In contrast to neonatal neurons, all of the young adult and aged neurons showed only a transient increase of [Ca2+]i in response to caffeine independent of nerve growth factor treatment. There was no significant effect of nerve growth factor on K+ depolarization-induced [Ca2+]i elevations at any of the ages studied. Nerve growth factor did not substantially alter the pattern of the transients induced by these three agents. Our results indicate that exogenous nerve growth factor is necessary to maintain normal acetylcholine receptor-mediated [Ca2+]i responses as well as Ca(2+)-induced Ca2+ release from intracellular calcium storage in neonatal superior cervical ganglion neurons. In mature superior cervical ganglion neurons, Ca2+ homeostasis becomes independent of exogenous nerve growth factor, and Ca2+ homeostasis and its independency are well preserved in aged neurons.

Aging↗

PubMed1998

Cyclic AMP-elevating agents prevent oligodendroglial excitotoxicity.

Previously, we have demonstrated that cells of the oligodendroglial lineage express non-NMDA glutamate receptor genes and are damaged by kainate-induced Ca2+ influx via non-NMDA glutamate receptor channels, representing oligodendroglial excitotoxicity. We find in the present study that agents that elevate intracellular cyclic AMP prevent oligodendroglial excitotoxicity. After oligodendrocyte-like cells, differentiated from the CG-4 cell line established from rat oligodendrocyte type-2 astrocyte progenitor cells, were exposed to 2 mM kainate for 24 h, cell death was evaluated by measuring activity of lactate dehydrogenase released into the culture medium. Released lactate dehydrogenase increased about threefold when exposed to 2 mM kainate. Kainate-induced cell death was prevented by one of the following agents: adenylate cyclase activator (forskolin), cyclic AMP analogues (dibutyryl cyclic AMP and 8-bromo-cyclic AMP), and cyclic AMP phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine, pentoxifylline, propentofylline, and ibudilast). Simultaneous addition of both forskolin and phosphodiesterase inhibitors prevented the kainate-induced cell death in an additive manner. A remarkable increase in Ca2+ influx (approximately 5.5-fold) also was induced by kainate. The cyclic AMP-elevating agents caused a partial suppression of the kainate-induced increase in Ca2+ influx, leading to a less prominent response of intracellular Ca2+ concentration to kainate. The suppressing effect of forskolin on the kainate-induced Ca2+ influx was partially reversed by H-89, an inhibitor of cyclic AMP-dependent protein kinase. In contrast to this, okadaic acid, an inhibitor of protein phosphatases 1 and 2A, brought about a decrease in the kainate-induced Ca2+ influx. We therefore concluded that cyclic AMP-elevating agents prevented oligodendroglial excitotoxicity by cyclic AMP-dependent protein kinase-dependent protein phosphorylation, resulting in decreased kainate-induced Ca2+ influx.

Animals↗

PubMed1998

AMPA receptor-mediated excitotoxicity in human NT2-N neurons results from loss of intracellular Ca2+ homeostasis following marked elevation of intracellular Na+.

Human NT2-N neurons express Ca2+-permeable alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid glutamate receptors (AMPA-GluRs) and become vulnerable to excitotoxicity when AMPA-GluR desensitization is blocked with cyclothiazide. Although the initial increase in intracellular Ca2+ levels ([Ca2+]i) was 1.9-fold greater in the presence than in the absence of cyclothiazide, Ca2+ entry via AMPA-GluRs in an early phase of the exposure was not necessary to elicit excitotoxicity in these neurons. Rather, subsequent necrosis was caused by a >40-fold rise in [Na+]i, which induced a delayed [Ca2+]i rise. Transfer of the neurons to a 5 mM Na+ medium after AMPA-GluR activation accelerated the delayed [Ca2+]i rise and intensified excitotoxicity. Low-Na+ medium-enhanced excitotoxicity was partially blocked by amiloride or dizocilpine (MK-801), and completely blocked by removal of extracellular Ca2+, suggesting that Ca2+ entry by reverse operation of Na+/Ca2+ exchangers and via NMDA glutamate receptors was responsible for the neuronal death after excessive Na+ loading. Our results serve to emphasize the central role of neuronal Na+ loading in AMPA-GluR-mediated excitotoxicity in human neurons.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1998

Hypoxic cell death in human NT2-N neurons: involvement of NMDA and non-NMDA glutamate receptors.

Human NTera2 teratocarcinoma cells were differentiated into postmitotic NT2-N neurons and exposed to hypoxia for 6 h. The cultures were evaluated microscopically, and percent lactate dehydrogenase (LDH) release after 24 and 48 h was used as an assay for cell death. After 48 h LDH release was 24.3 +/- 5.6% versus 13.8 +/- 3.7% in controls (p < 0.001). Cell death was greatly diminished by MK-801 pretreatment (15.4 +/- 5.1%, p < 0.001). If glutamine was omitted from the medium, glutamate levels after 6 h of hypoxia were reduced from 101 +/- 63 to 2.3 +/- 0.3 microM, and cell death at 48 h was also markedly reduced (15.4 +/- 4.5%, p < 0.001). The alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (18.7 +/- 5.1%, p < 0.001) and mild hypothermia (33.5-34 degrees C) during hypoxia (19.5 +/- 2.7%, p < 0.05) were moderately protective. Basic fibroblast growth factor (24.1 +/- 3.2%), the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (22.8 +/- 8.1%), the antioxidant N-tert-butyl-o-phenyinitrone (18.9 +/- 5.9%), and the 21-aminosteroid U74389G (24.0 +/- 3.4%) did not protect the cells. N-Acetyl-L-cysteine even tended to increase cell death (30.1 +/- 2.5%, p = 0.06). Treatment with MK-801 at the end of hypoxia did not reduce cell death (23.3 +/- 2.3%). In separate experiments, a 15-min exposure to 1 mM glutamate without hypoxia did not result in significant cell death (14.7 +/- 2.4 vs. 12.2 +/- 2.1%, p = 0.07). We conclude that, although somewhat resistant to glutamate toxicity when normoxic, NT2-N neurons die via an ionotropic glutamate receptor-mediated mechanism when exposed to hypoxia in the presence of glutamate. As far as we know, this is the first reported analysis of the mechanism of hypoxic cell death in cultured human neuronlike cells.

Cell Death↗

PubMed1997

Immunocytochemical expression of human muscle cell p75 neurotrophin receptor is down-regulated by cyclic adenosine 3',5'-monophosphate.

To investigate whether the immunocytochemical expression of low affinity neurotrophin receptor (p75) in human muscle is modulated by increased levels of intracellular cyclic adenosine 3',5'-monophosphate (cAMP), human cultured myogenic cells were treated with cAMP analogues dibutyryl cAMP (dbcAMP 0.5-1 mM) and 8-bromo cAMP (1 mM) or the adenylate cyclase activator forskolin (10-100 microM). Cultures were processed for indirect immunofluorescence microscopy using an anti-human p75 mAb. The treatment of cultured muscle cells with cAMP analogues or forskolin for two days induced a decrease of immunoreactivity for p75 and a reduction of both myotube formation and morphological cell differentiation. Removal of cAMP derivatives from the medium resulted in a return of immunoreactive cells to the levels of untreated controls. These data indicate that adenylate cyclase is involved in the regulation of human muscle p75.

Adenylyl Cyclases↗

PubMed1997

Expression of glutamic acid decarboxylase during human neuronal differentiation: studies using the NTera-2 culture system.

Human NTera-2N neurons, but not the parental NTera-2 teratocarcinoma line, decarboxylate [2-(15)N]glutamine to form gamma-[15N]aminobutyric acid (GABA). The reverse transcriptase-polymerase chain reaction (RT-PCR) followed by Southern blotting showed that NTera-2N neurons transcribe the glutamic acid decarboxylase p67 (GAD67) gene, and also demonstrated that there is developmentally regulated alternative splicing of GAD67 mRNA in NTera-2N neurons. As in rat central nervous system (CNS), this mRNA processing generates two RNA transcripts, owing to the inclusion or exclusion of an approximately 80 bp coding region insert. In embryonic day 16 (E16) rat brain, the larger of the two GAD67 mRNAs, which encodes a truncated, inactive apoenzyme, reaches a concentration almost equal to that of the smaller transcript, which encodes functional GAD67. In developing NTera-2N neurons, however, the larger transcript is barely detectable by RT-PCR. RT-PCR also revealed that rat CNS of all ages examined contains GAD65 mRNA, and that GAD65 mRNA is below the detectable range in NTera-2N neurons.

Animals↗

PubMed1997

Quantification of the effects of astrocytes on oligodendroglial morphology.

Oligodendrocytes in neonatal rat forebrain cultures grow either in isolation of other cells or upon astrocytes. Populations of both types of oligodendrocytes were used to quantify the effects of astrocytes on oligodendroglial morphology. Changes in oligodendroglial size and shape were determined by measurement of total process length, cell area, growth area, and fractal dimension. The directionality of process growth, quantified by measurement of the axes of growth, was also compared. Isolated oligodendrocytes exhibited greater total process length, greater cellular area, larger growth area, and a more complex boundary than oligodendrocytes growing upon astrocytes. Analysis of the axes of cellular growth revealed that the processes of isolated oligodendrocytes exhibited radial symmetry, whereas the processes of oligodendrocytes growing upon astrocytes were limited to an area demarcated by the astrocytic processes. These data suggest that, in neuron-free culture, the growth of oligodendroglial processes is modified by underlying astrocytic processes.

Animals↗

PubMed1997

Basic fibroblast growth factor prevents cAMP-induced apoptosis in cultured Schwann cells.

Peripheral nerve axotomy induces apoptosis in Schwann cell precursors; basic fibroblast growth factor (bFGF) protects these cells from axotomy-induced apoptosis (Jessen et al.: Neuron 12:509-527, 1994; Gavrilovic et al.: Eur J Neurosci 7:7-85, 1995). In this study, we investigate the effects of bFGF on apoptosis in neuron-free cultures of neonatal rat Schwann cells. Apoptotic cell death was induced in primary and secondary expanded Schwann cells by treatment with 1 mM concentrations of 8-bromoadenosine 3':5'-cyclic monophosphate (8-bromo-cAMP), a membrane-permeable analogue of cAMP which induces expression of galactocerebroside in the plasma membranes of Schwann cells. Treatment with bFGF reduced the percentage of galactocerebroside-bearing Schwann cells undergoing cAMP-induced DNA fragmentation. These findings suggest that bFGF can enhance the survival of terminally differentiated Schwann cells by preventing apoptosis.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1996

Pathophysiology of oligodendroglial excitotoxicity.

Oligodendrocyte-like cells (OLD) derived from the rat oligodendroglial precursor line, CG-4, express Ca(2+)-permeable non-methyl-D-aspartate glutamate receptor channels (GluR). Exposure to kainate, an L-glutamate analogue, markedly elevates OLC Ca2+ influx and cytosolic [Ca2+], and results in damage to both OLC plasma membrane and OLC nuclear DNA. Two observations indicate that kainate-induced OLC internucleosomal DNA nicking is not simply a delayed consequence of cell necrosis: 1) there is no temporal lag between onset of plasma membrane injury and of DNA nicking; and 2) aurintricarboxylic acid, an endonuclease inhibitor, blocks kainate-induced damage to the plasma membrane. N-acetyl-L-cysteine also inhibits OLC kainate injury, suggesting that reactive oxygen species participate in OLC excitotoxicity. Kainate-induced OLC Ca2+ influx and excitotoxicity are blocked by alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA), indicating that these kainate effects are mediated by AMPA-GluR. AMPA and L-glutamate fail to elicit OLC damage unless cyclothiazide, an AMPA-GluR desensitization blocker, is present. OLC express both the "flip" and "flop" forms of GluR2, GluR3, and GluR4 mRNAs, but neither flip nor flop GluR1 mRNA. These data, together with the restriction of the desensitization-blocking activity of cyclothiazide to GluR containing flip-encoded GluR subunits, and the sharply diminished Ca2+ permeability of GluR containing edited GluR2, suggest OLC excitotoxicity is mediated by AMPA-GluR that contain flip GluR3 and/or flip GluR4 protein subunits, but neither flip nor flop GluR2 protein subunits. Rapid desensitization of these GluR is likely to be important in protecting cells of the oligodendroglial lineage from excitotoxicity.

Acetylcysteine↗

PubMed1996

Re-entry into the cell cycle is required for bFGF-induced oligodendroglial dedifferentiation and survival.

Remyelination in the CNS following demyelinating disease may be accomplished by surviving mature oligodendrocytes that dedifferentiate, proliferate, migrate, and finally regenerate myelin. We previously reported that basic fibroblast growth factor (bFGF) induces oligodendrocytes in primary mixed glial cultures to dedifferentiate and synthesize DNA (Grinspan et al.: J Neurosci Res 36:672-680, 1993). We now show that this effect is direct and not mediated through the effects of bFGF on other cell types, because we were able to demonstrate similar changes in oligodendrocyte phenotype in enriched oligodendrocyte cultures prepared by immunopanning. The bFGF-induced reversion to the precursor stage of the oligodendroglial lineage can be blocked by agents that inhibit entry to the cell cycle; thus oligodendroglial dedifferentiation is dependent on proliferation. We also report that 2 days of bFGF treatment inhibits oligodendroglial apoptosis. However, when oligodendroglia are prevented from entering the cell cycle in the presence of bFGF, apoptotic cell death is increased. Thus, bFGF induces oligodendroglial dedifferentiation if oligodendroglial DNA synthesis can occur but causes oligodendroglial apoptosis when oligodendroglial DNA synthesis is prevented.

Animals↗

PubMed1996

Expression of N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptor genes in neuroblastoma, medulloblastoma, and other cells lines.

We evaluated expression of N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptor (GluR) genes by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blotting in nine established cell lines: rat CG-4 (oligodendroglial lineage) and RINm5F insulinoma cells; human CHP134, SMS-KCNR, SKNSH, and Nb69 neuroblastoma cells; and human D384Med, D425Med, and D458Med medulloblastoma cells. CG-4 expressed mRNAs encoding GluR2-7, KA-1, and KA-2 non-NMDA GluR (Yoshioka et al.: J Neurochem 64:2442-2448, 1995) and NR1 (NMDAR1) and NR2D NMDA GluR. After differentiation to oligodendrocyte-like cells, CG-4 also expressed NR2B mRNA. Rat insulinoma cells expressed GluR5 and KA-2 non-NMDA and NR1 and NR2D NMDA GluR mRNAs. The four human neuroblastoma lines all expressed mRNAs encoding GluR2-4, 6, 7 and KA-1 non-NMDA and NR1 NMDA GluR, and the three human medulloblastoma cell lines all expressed mRNAs encoding GluR1, 6 and KA-1, but none of the NMDA GluRs. Whereas CG-4 is susceptible to kainate excitotoxicity, treatment of insulinoma, neuroblastoma, and medulloblastoma lines with L-glutamate, kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), or NMDA failed to cause cell damage or to augment 45Ca2+ influx. Thus, despite expressing a variety of non-NMDA and NMDA GluR genes, the human neuroblastoma and medulloblastoma and rat insulinoma lines failed to assemble Ca(2+)-permeable NMDA or non-NMDA GluR channels. This failure confers protection against excitotoxicity and may contribute to progression of tumors of these types.

Animals↗

PubMed1996

Human neural tissues express a truncated Ror1 receptor tyrosine kinase, lacking both extracellular and transmembrane domains.

Human heart, lung and kidney express a 6 kb mRNA encoding Ror1, a member of the receptor tyrosine kinase (RTK) family with as yet unknown ligand specificity. We used a Ror1 cDNA probe to screen a cDNA library prepared from the human neuronogenic teratocarcinoma line, NTera2, and cloned a 2373 nucleotide transcript. This transcript contains an open reading frame that encodes a 388 amino acid protein identical with the cytosolic, C-terminal region of ror1 but lacking the ror1 transmembrane and entire extracellular domains. Northern blots demonstrate that mRNA encoding this truncated Ror1 ('t-Rorl') is abundantly expressed in fetal and adult human CNS, in human leukemia, lymphoma cell lines, and in a variety of human cancers derived from neuroectoderm. While previous studies have documented alternative splicing patterns within 5' and 3' regions of mRNAs encoding various RTKs altering their ligand binding specificity or their intracellular signaling, the present report is the first to demonstrate tissue-specific alternative mRNA splicing causing loss of the entire extracellular and transmembrane regions of an RTK.

Base Sequence↗

PubMed1996

Complexity analysis of oligodendroglial processes expressing myelin-associated glycoprotein.

Oligodendroglia synthesize myelin in the mammalian central nervous system. Mature oligodendroglia have been identified in culture by two criteria; the expression of molecules characteristic of myelin, such as galactocerebroside (galC) and myelin-associated glycoprotein (MAG), and the elaboration of complex processes. Myelin gene expression can be documented by the binding of specific antibodies and antisera to the myelin-specific molecules; process complexity can be described by the fractal dimension, D. In this study, anti-MAG antisera was used to document MAG expression in the processes of oligodendroglia. Eighty percent of the galC+ oligodendroglia bound anti-MAG antiserum. With time in culture, MAG immunoreactivity seemed to extend from the cell soma into the oligodendroglial processes. To quantify this observation, fractal dimensions were calculated using either galC or MAG immunoreactivity to visualize oligodendroglial processes. A fractal dimension of 1.5 was calculated for O1+ processes by day 4 of culture; this value for D remained constant over the course of 1 month in culture. The fractal dimension calculated for MAG+ processes increased from 1.2 to 1.5 over the course of 28 days in culture. This change in fractal dimension confirms our visual impression that galC-containing processes acquire MAG slowly over the course of several weeks in culture.

Animals↗

PubMed1996

Astrocyte leucine metabolism: significance of branched-chain amino acid transamination.

We studied astrocytic metabolism of leucine, which in brain is a major donor of nitrogen for the synthesis of glutamate and glutamine. The uptake of leucine into glia was rapid, with a Vmax of 53.6 +/- 3.2 nmol/mg of protein/min and a Km of 449.2 +/- 94.9 microM. Virtually all leucine transport was found to be Na+ independent. Astrocytic accumulation of leucine was much greater (3x) in the presence of alpha-aminooxyacetic acid (5 mM), an inhibitor of transamination reactions, suggesting that the glia rapidly transaminate leucine to alpha-ketoisocaproic acid (KIC), which they then release into the extracellular fluid. This inference was confirmed by the direct measurement of KIC release to the medium when astrocytes were incubated with leucine. Approximately 70% of the leucine that the glia cleared from the medium was released as the keto acid. The apparent Km for leucine conversion to extracellular KIC was a medium [leucine] of 58 microM with a Vmax of approximately 2.0 nmol/mg of protein/min. The transamination of leucine is bidirectional (leucine+alpha-ketoglutarate<==>KIC+glutamate) in astrocytes, but flux from leucine-->glutamate is more active than that from glutamate-->leucine. These data underscore the significance of leucine handling to overall brain nitrogen metabolism. The release of KIC from glia to the extracellular fluid may afford a mechanism for the "buffering" of glutamate in neurons, which would consume this neurotransmitter in the course of reaminating KIC to leucine.

Amination↗

PubMed1996

Induction of apoptosis by La Crosse virus infection and role of neuronal differentiation and human bcl-2 expression in its prevention.

La Crosse virus causes a highly cytopathic infection in cultured cells and in the murine central nervous system (CNS), with widespread neuronal destruction. In some viral infections of the CNS, apoptosis, or programmed cell death, has been proposed as a mechanism for cytopathology (Y. Shen and T. E. Shenk, Curr. Opin. Genet. Dev. 5:105-111, 1995). To determine whether apoptosis plays a role in La Crosse virus-induced cell death, we performed experiments with newborn mice and two neural tissue culture models. Newborn mice infected with La Crosse virus showed evidence of apoptosis with the terminal deoxynucleotidyl transferase-mediated nicked-end labeling (TUNEL) assay and, concomitantly, histopathological suggestion of neuronal dropout. Infection of tissue culture cells also resulted in DNA fragmentation, TUNEL reactivity, and morphological changes in the nuclei characteristic of apoptotic cells. As in one other system (S. Ubol, P. C. Tucker, D. E. Griffin, and J. M. Hardwick, Proc. Natl. Acad. Sci. USA 91:5202-5206, 1994), expression of the human proto-oncogene bcl-2 was able to protect one neuronal cell line, N18-RE-105, from undergoing apoptosis after La Crosse virus infection and prolonged the survival of infected cells. Nevertheless, expression of bcl-2 did not prevent eventual cytopathicity. However, a human neuronal cell line, NT2N, was resistant to both apoptosis and other types of cytopathicity after infection with La Crosse virus, reaffirming the complexity of cell death. Our results show that apoptosis is an important consequence of La Crosse virus infection in vivo and in vitro.

Animals↗

PubMed1995

Low-affinity nerve growth factor receptor expression in sciatic nerve during P2-peptide induced experimental allergic neuritis.

Northern blot, immunocytochemistry, and single nerve fiber immunostaining were used to determine the expression of low-affinity nerve growth factor (NGF) receptor (p75NGFR) in the peripheral nervous system of Lewis rats during the course of experimental allergic neuritis (EAN) induced with 100 micrograms of 'SP26' synthetic peptide, corresponding to residues 53-78 of bovine P2 myelin protein. This severe reversible polyneuropathy, characterized by extensive multifocal demyelination in nerve roots, and axonal degeneration within sciatic nerve, represents a useful tool to investigate Schwann cell gene expression in these different pathological conditions. Our results showed that both p75NGFR mRNA and protein were induced within sciatic nerve by day 18 and 23 after 'SP26' immunization. By this time, however, p75NGFR was not detectable in roots of cauda equina. These data demonstrate that, during EAN induced by immunization with 100 micrograms of 'SP26', p75NGFR is up-regulated in Schwann cells and in nerve fibers distally along the nerve. This suggests that the induction of the receptor in sciatic nerve during P2-peptide EAN is related to the loss of axonal contact by Schwann cells.

Animals↗

PubMed1995

Apoptosis occurs in the oligodendroglial lineage, and is prevented by basic fibroblast growth factor.

During the perinatal period, oligodendroglial precursor cells proliferate rapidly, then cease dividing and differentiate into oligodendroglia. Many of these newly formed oligodendroglia are destined to die. We now demonstrate that oligodendroglia generated in passaged cultures of rat forebrain oligodendroglial precursor cells after removal of basic fibroblast growth factor (basic FGF) from the medium often undergo internucleosomal DNA nicking and nuclear fragmentation, features characteristic of apoptosis. These alterations are rare in cultures maintained continuously in basic FGF. As in many other cellular lineages susceptible to apoptosis, these degenerative changes can be prevented by treatment with the endonuclease antagonist, aurintricarboxylic acid, or by inhibiting de novo RNA or protein synthesis. Supplementation of the basic FGF-free medium with insulin, insulin-like growth factor-1, platelet-derived growth factor, or ciliary neuronotrophic growth factor also diminishes DNA nicking. Both oligodendroglial differentiation and DNA nicking are induced in basic FGF-treated cultures by inhibiting DNA synthesis with aphidicholin or excess thymidine, thus suggesting a close linkage between the anti-apoptotic, anti-differentiation, and mitogenic effects of basic FGF on the oligodendroglial lineage.

Animals↗