Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.22.61 (caspase-8)
 6,833 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have used C6 glial cells (2B clone), early and late passage, as well as advanced passages (8-17) of glial cells derived from aged (18-month-old) mouse cerebral hemispheres (MACH), as model systems for studying glial properties. In this study passages 20-24 were considered "early" and passages 73-90 were considered "late." Activities of glutamine synthetase (GS) and cyclic nucleotide phosphohydrolase (CNP) were used as biochemical markers for astrocytes and oligodendrocytes, respectively. Glial phenotypes were identified immunocytochemically using double staining for glial fibrillary acidic protein (GFAP) and A2B5 antigen (type 1 and type 2 astrocytes) or galactocerebroside (GalC) and A2B5 antigen (oligodendrocytes); cells positive for A2B5 and negative for both GFAP and GalC were considered to be precursor cells. Cultures were grown either in DMEM supplemented with 10% fetal bovine serum or in serum-free chemically defined medium (CDM) supplemented with insulin and transferrin. We report that early-passage C6 glial cells continue to be bipotential cells and when grown in the absence of serum express high GS and CNP activities correlating with the high number of GFAP- and GalC-positive cells, respectively. Late-passage cells continued to be committed to the type 2 astrocytic phenotype regardless of media composition (+/- serum). MACH cultures consist of protoplasmic type 1 astrocytes, differentiated type 2 astrocytes, and oligodendrocytes as well as glial progenitor cells. When these cultures were grown in CDM+transferrin, both GS and CNP activities increased, suggesting that transferrin has provided the signal for progenitor cells present in these cultures derived from aged brain to differentiate into type 2 astrocytes and oligodendrocytes.
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PMID:Comparative biochemical, morphological, and immunocytochemical studies between C-6 glial cells of early and late passages and advanced passages of glial cells derived from aged mouse cerebral hemispheres. 136 Nov 80

We have previously reported that glial cells derived from aged mouse cerebral hemispheres (MACH) in primary cultures and after several passages consist of protoplasmic astrocytes (Type 1), differentiated stellate astrocytes (Type 2), a few oligodendrocytes, and also glial precursors. In this study, we examined the influence of culture substrata: plastic, poly-L-lysine, laminin or collagen on the differentiation of MACH glial cells of advanced passages (P18-19) using glutamine synthetase (GS) and cyclic nucleotide phosphohydrolase (CNP) activity as biochemical markers for astrocytes and oligodendrocytes, respectively. Cultures were also examined morphologically using light microscopy. In general, GS activity was increased in cultures grown on the three chemical substrata versus plastic alone with the most striking effect being the 2-fold increase observed in those cells grown in laminin. No differences were noted in CNP activity. Morphologically, proliferation of protoplasmic (Type 1) astrocytes was enhanced by culture day 2 on polylysine substratum and stellate differentiated (Type 2) astrocytes were noted on collagen. The striking feature in cultures grown on laminin was the presence of astrocytes with markedly long processes. Thus, morphological astrocyte differentiation appears to correspond to the increased GS activity. We propose that the extracellular matrix components such as collagen and laminin may play an important role in promoting glial precursors to differentiate into astrocytes.
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PMID:Influence of culture substrata on the differentiation of advanced passage glial cells in cultures from aged mouse cerebral hemispheres. 790 67

In this study we used as glial cell models, early and late passage C-6 glial cells, 2B clone, and advanced passages of glial cells derived from aged mouse cerebral hemispheres (MACH) to examine responsiveness to opioids. We have previously reported that early passage C-6 glial cells, 2B clone, are bipotential and can be geared toward oligodendrocyte or astrocytic expression, whereas late passage C-6 glial cells are astrocytic. In addition, MACH cultures have been previously characterized and consist of astrocytes type 1 and 2, some oligodendrocytes, and few glial precursors. In this study, early passage (17-20) and late passage (106-108) C-6 glial cells or MACH cells of passages 16-19 were grown from plating time until harvesting, day 7 or 8, in DMEM + 10% FBS in the presence or absence of opioid peptides, Leu-enkephalin (10(-8) to 10(-10) M) or its synthetic analog, dalargin (Tyr-D-Ala-Gly-Phe-Leu-Arg; 10(-8) to 10(-10) M). We examined for the activities of glutamine synthetase (GS) and cyclic nucleotide phosphohydrolase (CNP), enzyme markers for astrocytes and oligodendrocytes, respectively. We found that CNP activity was markedly increased in the early passage following opioid treatment, indicative of a shift to oligodendrocytic expression. In the late passage cells, already committed to astrocytic expression, opioid treatment enhanced GS activity suggesting that astrocytes respond to opioids. GS activity was markedly increased in MACH cultures grown in the presence of opioids with no changes in CNP. Thus, type 1 astrocytes, the predominant glial type in MACH cultures, responded to opioids.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Maintenance of glial plasticity with aging in C-6 glial cells and normal astrocytes in culture: responsiveness to opioid peptides. 790 40

In this study, we were interested to compare the responsiveness to growth factors, NGF, b-FGF and EGF and cytokines, IL1 beta, and TNF-alpha, in late passages (74-79) C6 glial cells committed astrocytes and astrocytes of advanced passages (26-28) in cultures derived from aged mouse cerebral hemispheres (MACH). Cultures were grown in either DMEM or chemically defined medium (CDM/TIPS) in order to test the effects of growth factors or cytokines. The activity of glutamine synthetase (GS), a marker for astrocytes, was used as a test parameter. We found that treatment with growth factors increased GS activity in both glial cell culture systems with the exception of EGF in C-6 glial cells. Treatment with cytokines markedly decreased GS activity in the late passage C6 glial cells whereas only TNF-alpha had a similar effect on MACH astrocytes. In view of the generally opposite effects of growth factors and cytokines on GS activity, we speculate that these molecules which are also endogenously present in glial cells may play a role in the maintenance of cellular homeostasis.
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PMID:Differential responsiveness of late passage C-6 glial cells and advanced passages of astrocytes derived from aged mouse cerebral hemispheres to cytokines and growth factors: glutamine synthetase activity. 872 70

We have been using glial cells derived from aged mouse cerebral hemispheres (MACH) at several passages to study the responsiveness of astrocytes to microenvironmental signals in culture. In the present study, we examined the effects of excitatory amino acids on the activity of glutamine synthetase, a marker for astrocytes. MACH glia cell passages 25 to 29 were used. Culture groups were Dulbecco's modified Eagle's medium +10% fetal bovine serum (control); glutamate 100 microM; gamma-amino-3-hydroxy-5-methyl isoxazole-4-propionic acid (AMPA) 50 microM; kainic acid 10 microM; N-methyl-D-aspartate (NMDA) 10 microM. In all treated groups glutamine synthetase activity was significantly higher than in controls. We speculate that this increase represents an enhanced differentiation of immature astrocytes. In a second series, we examined the effects of glutamate receptor antagonists on glutamine synthetase activity as follows. MACH cultures were treated with glutamate 100 microM in combinations with either L(+)-2-amino-3-phosphonopropionic acid (L-AP3; 50 microM); D(-)-2-amino-5-phosphonopentanoic acid (D-AP5; 50 microM) or 6,7-dinitroquinoxaline-2,3-dione (DNQX, 50 microM). The increase in GS activity produced by glutamate was inhibited by the non-selective NMDA receptor antagonist, DNQX, but not by the metabotropic receptor antagonist, L-AP3 or a selective NMDA receptor antagonist, D-AP5. We also found that in cultures treated with glutamate, a number of astrocytes resembled "reactive astrocytes" morphologically. These astrocytes were absent in cultures treated with glutamate+DNQX. The findings provide supportive evidence that astrocytes from aged mouse cerebral hemispheres respond to excitatory amino acids and that this response is mediated by non-NMDA receptor activation.
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PMID:Stimulation of glutamine synthetase activity by excitatory amino acids in astrocyte cultures derived from aged mouse cerebral hemispheres may be associated with non-N-methyl-D-aspartate receptor activation. 888 86

This study was targeted at the beginning to understand the functional status of glial cells derived from aged brain. We have previously characterized passaged cell cultures derived from aged mouse cerebral hemispheres (MACH) and found them to contain large populations of astrocytes, type 1, as well as limited numbers of astrocytes, type 2, oligodendrocytes, and progenitor cells. Using the activity of the astrocyte marker, glutamine synthetase (GS), as an index, we found that MACH astrocytes continue to respond to several microenvironmental signals, including the cAMP-enhancing agents dibutyryl cAMP and R020-1724 (an inhibitor of phosphodiesterase). In addition, whereas the basal activity of GS increased with cell passage, their response to these agents was cell-passage dependent, increasing at early (21-22) passages and decreasing at later (46-51) passages. Because neurotrophins (i.e., NGF and EGF) also provide microenvironmental signals essential to normal glial function, MACH cultures were assessed for their response to these factors. MACH cultures at passage 35 responded to treatment with NGF and EGF with a dose-dependent increase in GS activity by both neurotrophins. With the intention of arresting these cultures at a specific stage of differentiation, these cells were immortalized at passage 19 by transfection with the gene encoding SV40 Large T antigen. These immortalized MACH responded to exposure to dBcAMP and RO20-1724 with a marked decrease in GS activity, mimicking the response of normal MACH glia at late passage. Finally, because it has been shown that glia from both immature and adult brain contain neurotrophins and respond to neurotrophins via a receptor-mediated pathway, we examined expression of NGF protein as well as NGF (p75) and EGF receptor protein in various passages and colonies of normal and immortalized MACH cultures. We found a consistent expression of all three proteins in the various cell populations. Results of this study suggest that astrocytes from aging brain continue to function normally with respect to several parameters (i.e., response to neurotrophins and differentiating agents). Thus, they retain their plasticity to a great degree through early cell passages. However, with advancing cell passage this plasticity declines and cell homeostasis is impaired. We propose, therefore, that astrocytes undergo several critical periods in their functional lifespan, one of which is represented by the functional transition demonstrated in this study.
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PMID:Plasticity of astrocytes derived from aged mouse cerebral hemispheres: changes with cell passage and immortalization. 896 86