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)

Neuron-glia cocultures were prepared using, as a source for glial cells, either C6 glia (2B clone) of early (2B23) or late (2B111) passages or advanced passages of glial cells derived from primary cultures prepared from aged mouse cerebral hemispheres (MACH). Six-day-old chick embryo cerebral hemispheres (E6CH) were the source of neuron-enriched cultures. Glutamine synthetase (GS) activity was used as a marker for astrocytes and 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP) activity was used as a marker for oligodendrocytes. GS activity was markedly enhanced in cocultures of E6CH neurons and 2B23 glioblastic cells, whereas GS activity was reduced in cocultures of E6CH neurons and 2B111 astrocytic glia. In contrast, CNP activity was enhanced in cocultures of C6 glial cells with E6CH neurons. Glial cells from aged mouse brain did not respond to coculturing with E6CH neurons. It appears from these findings that neuronal input enhances the differentiation of glioblastic cells to either astrocytic or oligodendrocytic expression, whereas it decreases the activity of committed astrocytes. In contrast, glial cells from aged mouse brain do not respond to neuronal input. Choline acetyltransferase (ChAT) activity, a marker for cholinergic neurons, was enhanced only when E6CH cultures were grown in conditioned medium (CM) from 2B23 glioblastic cells. In contrast, ChAT activity was markedly diminished when E6CH neurons were cocultured with MACH glial cells but not when grown in CM from MACH glial cells. Thus, humoral factors from immature glial cells appear to enhance cholinergic neuronal phenotypic expression whereas cell-cell membrane contacts with aged glial cells diminish cholinergic phenotypic expression. The findings present supportive evidence that neuron-glia interrelationships are age dependent.
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PMID:Differences in neuronal and glial cell phenotypic expression in neuron-glia cocultures: influence of glia-conditioned media and living glial cell substrata. 135 4

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 earlier studies, we established glial cell cultures derived from aged (18-month-old) mouse cerebral hemispheres (MACH) and have maintained them frozen at various passages. These cultures were characterized immunocytochemically and consist of: 5% oligodendrocytes (GalC+), 75% astrocytes-type 1 (GFAP+ only), 15% astrocytes-type 2 (GFAP+ + A2B5+), and 5% progenitor glial cells (A2B5+ only). In the present study, we isolated colonies from MACH passage 29 cultures and also colonies from MACH passage 19 transfected with the gene for SV40 large T antigen and further subcultured for 8 passages. Using double-staining immunocytochemistry, we found in non-transfected MACH passage 19 colonies consisting primarily of cells exhibiting only vimentin-positive staining and are considered to be immature glioblasts; colonies consisting primarily of cells exhibiting GFAP+ + vimentin+ which are considered to be astrocytes at an intermediate stage of maturation; and colonies consisting predominantly of cells exhibiting GFAP+ only which are considered to be mature astrocytes. In contrast, colonies isolated from transfected MACH cultures consisted primarily of vimentin+ cells. In conclusion, astrocytes in cultures derived from aged brain continue to be variable as they are during development. However, their response to the microenvironment may differ during development and during aging. Thus, the availability of clones of mature and immature astrocytes offers the opportunity to study neuron-glia interactions and the role of mature and immature astrocytes in neuronal aging and regeneration.
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PMID:Glial cells derived from aged mouse brain in culture display both mature and immature astrocytic phenotypes. 793 75

APO-1 (Fas/CD95), a member of the tumor necrosis factor receptor superfamily, induces apoptosis upon receptor oligomerization. In a search to identify intracellular signaling molecules coupling to oligomerized APO-1, several cytotoxicity-dependent APO-1-associated proteins (CAP) were immunoprecipitated from the apoptosis-sensitive human leukemic T cell line HUT78 and the lymphoblastoid B cell line SKW6.4. CAP1-3 (27-29 kDa) and CAP4 (55 kDa), instantly detectable after the crosslinking of APO-1, were associated only with aggregated (the signaling form of APO-1) and not with monomeric APO-1. CAP1 and CAP2 were identified as serine phosphorylated MORT1/FADD. The association of CAP1-4 with APO-1 was not observed with C-terminally truncated non-signaling APO-1. In addition, CAP1 and CAP2 did not associate with an APO-1 cytoplasmic tail carrying the lprcg amino acid replacement. Moreover, no APO-1-CAP association was found in the APO-1+, anti-APO-1-resistant pre-B cell line Boe. Our data suggest that in vivo CAP1-4 are the APO-1 apoptosis-transducing molecules.
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PMID:Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signaling complex (DISC) with the receptor. 852 15

Fas/APO-1 and p55 tumor necrosis factor (TNF) receptor (p55-R) activate cellular mechanisms that result in cell death. Upon activation of these receptors, Fas/APO-1 binds a protein called MORT1 (or FADD) and p55-R binds a protein called TRADD. MORT1 and TRADD can also bind to each other. We have cloned a novel protein, MACH, that binds to MORT1. This protein exists in multiple isoforms, some of which contain a region that has proteolytic activity and shows marked sequence homology to proteases of the ICE/CED-3 family. Cellular expression of the proteolytic MACH isoforms results in cell death. Expression of MACH isoforms that contain an incomplete ICE/CED-3 region provides effective protection against the cytotoxicity induced by Fas/APO-1 or p55-R triggering. These findings suggest that MACH is the most upstream enzymatic component in the Fas/APO-1- and p55-R-induced cell death signaling cascades.
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PMID:Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1- and TNF receptor-induced cell death. 868 76

To identify CAP3 and CAP4, components of the CD95 (Fas/APO-1) death-inducing signaling complex, we utilized nano-electrospray tandem mass spectrometry, a recently developed technique to sequence femtomole quantities of polyacrylamide gel-separated proteins. Interestingly, CAP4 encodes a novel 55 kDa protein, designated FLICE, which has homology to both FADD and the ICE/CED-3 family of cysteine proteases. FLICE binds to the death effector domain of FADD and upon overexpression induces apoptosis that is blocked by the ICE family inhibitors, CrmA and z-VAD-fmk. CAP3 was identified as the FLICE prodomain which likely remains bound to the receptor after proteolytic activation. Taken together, this is unique biochemical evidence to link a death receptor physically to the proapoptotic proteases of the ICE/CED-3 family.
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PMID:FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death--inducing signaling complex. 868 77

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

Emerging evidence suggests that an amplifiable protease cascade consisting of multiple aspartate specific cysteine proteases (ASCPs) is responsible for the apoptotic changes observed in mammalian cells undergoing programmed cell death. Here we describe the cloning of two novel ASCPs from human Jurkat T-lymphocytes. Like other ASCPs, the new proteases, named Mch4 and Mch5, are derived from single chain proenzymes. However, their putative active sites contain a QACQG pentapeptide instead of the QACRG present in ail known ASCPs. Also, their N termini contain FADD-like death effector domains, suggesting possible interaction with FADD. Expression of Mch4 in Escherichia coli produced an active protease that, like other ASCPs, was potently inhibited (Kj = 14 nM) by the tetrapeptide aldehyde DEVD-CHO. Interestingly, both Mch4 and the serine protease granzyme B cleave recombinant proCPP32 and proMch3 at a conserved IXXD-S sequence to produce the large and small subunits of the active proteases. Granzyme B also cleaves proMch4 at a homologous IXXD-A processing sequence to produce mature Mch4. These observations suggest that CPP32 and Mch3 are targets of mature Mch4 protease in apoptotic cells. The presence of the FADD-like domains in Mch4 and Mch5 suggests a role for these proteases in the Fas-apoptotic pathway. In addition, these proteases could participate in the granzyme B apoptotic pathways.
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PMID:In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains. 875 96


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