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:3.4.24.3 (collagenase)
18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Mouse collagenase cDNA was cloned and sequenced. The deduced amino acid sequence was compared to those of the other mammalian collagenases and related matrix metalloproteinases. These comparisons, as well as those of some enzymatic properties, show that the rodent (mouse and rat) interstitial collagenases are very similar but differ more from the other interstitial collagenases than does human neutrophil collagenase. This supports the hypothesis that the order Rodentia is an outgroup to the other eutherian (placental) mammalian orders.
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PMID:Cloning and sequencing of mouse collagenase cDNA. Divergence of mouse and rat collagenases from the other mammalian collagenases. 138 28

Extracellular, membrane-bound vesicles are widely regarded to be the initial site of calcification in a variety of tissues under normal and pathological conditions. Alkaline phosphatase is believed to play a vital role in this process by hydrolysing ester phosphates or mineral inhibitors, e.g. inorganic phosphates. In the present study, matrix vesicles from normal and rachitic rat growth plates were compared with regard to specific activity of alkaline phosphatase, total vesicle protein and ultrastructural distribution of alkaline phosphatase activity. Matrix vesicles were released from normal or rachitic growth plates by collagenase digestion and isolated by differential centrifugation. Enzyme cytochemical localization involving a cerium capture method was performed on vesicles collected by vacuum filtration on Millipore filters. SDS gels and Western blots on fractions of both normal and rachitic matrix vesicles showed major proteins to be almost identical and confirmed the presence of alkaline phosphatase in both. Total matrix vesicle protein ((mg total matrix vesicle protein/rat) x 10(2)) per rat was significantly greater for the rachitic animals (9.0 +/- 2.0 vs. 4.0 +/- 1.0), P less than 0.0001. Alkaline phosphatase specific activity (units alkaline phosphatase/mg vesicle protein) in the rachitic and normal matrix vesicles was 25.29 +/- 9.36 and 18.78 +/- 3.37, respectively (0.05 less than P less than 0.1). Electron dense cerium phosphate deposits were localized to the outer membrane surface of matrix vesicles derived from both types of rats. This data, the first to quantify the relationship between rickets, matrix vesicle protein and alkaline phosphatase specific activity, suggests that matrix vesicles from rachitic and normal rats have biochemical and morphological similarity.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Increased matrix vesicle protein in rachitic rat epiphyseal growth plates. 165 31

P-450IIB genes (P-450b and P-450e in rat) are induced following treatment with phenobarbital predominantly in hepatocytes located in zones 2 and 3 of the liver acinus. The previous finding of phenobarbital-mediated induction of P-450IIB mRNAs and apoproteins in the same zonal hepatocytes suggests that this differential induction is most likely due to zonal differences in the activation of gene transcription. To determine whether differential P-450IIB gene transcription was dependent on the type of inducer (i.e., inducer specificity) or on the capacity of hepatocytes located in different acinar zones to respond to inducers (i.e. zonal specificity), the pattern of acinar induction was evaluated following treatment with inducers that have diverse physiochemical properties and inductive capacities. The zonal distribution of P-450b and P-450e mRNAs in liver was determined by in situ hybridization after the administration of either phenobarbital, polychlorinated biphenyls, chlordane, or chlorpromazine. Liver sections were hybridized with 3H-labeled RNA transcripts of a P-450e cDNA that recognizes sequences of both P-450b and P-450e mRNAs and the pattern of zonal mRNA induction was measured by quantitative image analysis. Each inducer increased P-450b,e mRNA levels predominantly in hepatocytes of zones 2 and 3 of the hepatic acinus. The P-450b and P-450e apoproteins were induced in the same zonal hepatocytes as the P-450b,e mRNAs as shown by immunofluorescence studies using monoclonal antibodies. Therefore, differential transcriptional induction of the P-450IIB genes in the liver acinus does not seem to be dependent on a specific chemical inducer, but rather is a characteristic capacity of hepatocytes located in different acinar zones. To determine whether the induction of P-450b and P-450e was dependent on the liver tissue organization, hepatocytes were isolated by collagenase perfusion of the liver and transplanted into the spleens of syngeneic rats. Induction of P-450b and P-450e mRNAs and apoproteins, assessed 1 month after transplantation, was evident with each of the chemical inducers. These data suggest that, once hepatocytes have attained the capacity to respond to inducers, the organization of hepatocytes into acini and the sinusoidal microenvironment of the liver are not required for hepatocytes to maintain the ability to respond to inducers with an increase in transcription of P-450IIB genes. Moreover, the splenic hepatocytes demonstrated a heterogeneous pattern of P-450b,e apoprotein induction; this raises the possibility that the acinar organization is also not required for the heterogenous expression of P-450IIB genes.
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PMID:Induction of P-450IIB genes within the rat liver acinus is not dependent on the chemical inducer or on the acinar organization. 247 Jul 62

In order to standardize and to characterize a chondrocyte primary culture, cells from rat rib resting cartilage were used. High yield (0.99 +/- 0.18 x 10(6) cells/rat) and viability (91.76%) of costal cartilage cells was reached by enzymatic digestion with collagenase. The cells were cultivated in Dulbecco's medium (DME) supplemented with 10%. Heat inactivated newborn calf serum, at 37 degrees under humidified atmosphere of 5% CO2 in air. Two or three days after plating, the cells were attached to the surface of tissue culture weel, and began dividing. Adhesion was independent of plating density. The doubling time of cell population was found to be 23.19 hours. The cells became a monolayer and required easy maintenance. The results support the contention that rat costal cartilage is a good source of chondrocytes for primary culture cells experiments.
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PMID:[Characterization of a chondrocyte primary culture from rib cartilage of the rat]. 263 37

The development of a humoral immune response to the tubular basement membrane (TBM) alloantigen of Brown-Norway (BN) rat kidneys was studied after transplantation of BN rat kidneys into bilaterally nephrectomized Lewis (LEW) rats. The LEW rat recipients consisted of four groups receiving no form of immunosuppression, pretransplantation cyclosporin alone, or pretransplantation donor-specific or donor-nonspecific transfusions combined with cyclosporin. The latter two regimens induce indefinite allograft survival in the majority of recipients. Circulating antibody to collagenase-solubilized BN rat renal basement membrane (CS-BN-RBM) was present in all four groups of transplant recipients within 1 week after transplantation, and no significant differences in antibody levels were noted between rats receiving no immunosuppression (survival of 1-2 weeks) and the groups of rats who received various immunosuppressive regimens and survived longer. Circulating antibody to BN-CS-RBM continued to increase in quantity in the cyclosporin-treated group until the time of death (2-10 weeks post-transplantation). In the much longer lived combined transfusion and cyclosporin-treated groups, circulating antibody to BN-CS-RBM generally attained a maximum at approximately 2 to 4 months post-transplantation and then plateaued or decreased somewhat before the time of death (3-16 months post-transplantation). No correlation was found between quantity of circulating anti-BN-CS-RBM antibody and post-transplantation survival. Comparative study of the quantity of circulating antibody to BN-CS-RBM (the presumed nephritogenic antigen of experimental tubulointerstitial nephritis in the BN rat) in serum from transplant recipients as compared to serum from BN rats with severe experimental tubulointerstitial nephritis (TIN) (as induced by immunization with heterologous TBM antigens) demonstrated a greater quantity of potentially nephritogenic antibody circulating in transplant recipients than in BN rats with experimental TIN. Histologically, the transplanted kidneys in immunomodulated recipients demonstrated focal chronic interstitial inflammatory infiltrates with tubular atrophy and relative sparing of the glomeruli. The development of immune responses to tissue-specific alloantigens may become of clinical significance as graft-survival times are increased.
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PMID:The development of humoral immunity to tissue-specific tubular basement membrane alloantigens after renal transplantation across the major histocompatibility barrier in rats immunomodulated with blood transfusions and cyclosporin. 329 55

1. A method for measuring the lipogenesis from [(14)C]glucose by single fat cells is described: (i) after incubation with ;carrier-free' [U-(14)C]glucose (0.55 mu-mole/ml.), collagenase-isolated fat cells were fixed with osmium tetroxide; (ii) similarly incubated pieces of epididymal fat pads were treated with osmium tetroxide for 90 sec, whereby only the superficial cells are fixed, and the tissue was then disintegrated by shaking with collagenase. The osmium-fixed free cells were washed, sucked into a micropipette, measured under a microscope and assayed individually for (14)C-activity.2. There was a quantitative recovery of (14)C-lipid activity from osmium-fixed single cells.3. Both collagenase-isolated cells and in situ fixed surface cells were normally distributed with respect to diameters (for both cell groups from ad lib. fed rats of ca. 110 g; mean diameter, about 55 mum; S.D. about 7 mum).4. Frequency distribution curves (number of fat cells versus (14)C-lipogenesis per cell) were asymmetric and very broad (coefficient of variation about 50%) for collagenase-isolated cells incubated with insulin (10(3) mu-u./ml.). Frequency distribution curves for surface cells obtained from similarly incubated pieces of epididymal fat pads showed a coefficient of variation of the same magnitude, whereas the mean lipogenesis of these cells was only about one third of that of the isolated cells.5. Collagenase-isolated cells incubated in the presence of insulin (10(3) mu-u./ml.) showed a weak but highly significant positive correlation between fat cell diameter and (14)C-lipogenesis (eight rats, r about 0.5 and P < 0.001 for each rat). Analysis of the relationship: lipogenesis = k x diameter to the exponent of beta showed that the estimates of beta varied significantly from rat to rat (range: 1.3-2.9). Similar correlations between cell size and lipogenesis were found both for cells incubated with insulin in various submaximal concentrations and for cells incubated without insulin.6. Small and large cells from the same rat were equally sensitive to insulin.7. Statistical analysis of frequency distribution curves (number of cells versus (14)C-lipogenesis per unit surface area) representing cells from the same rat incubated with insulin 0, 2.5, 5, 10, and 10(3) mu-u./ml., respectively, suggests that insulin exerts a graded influence on the lipogenesis of each fat cell.
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PMID:Lipogenesis and insulin sensitivity of single fat cells. 436 2

A new method for preparing non-parenchymal rat liver cells (NPC) is described. The liver cell suspension, prepared by perfusing the liver with collagenase, was treated with enterotoxin from Clostridium perfringens for 15 min. The enterotoxin made the parenchymal cells leaky, and these cells could be separated from the NPC by centrifugation in a solution containing Nycodenz (20%, w/v). During the centrifugation, the NPC floated, while the parenchymal cells sedimented. The yield of NPC per liver (200 g rat) was about 250 X 10(6) cells. The NPC were further separated into endothelial cells, Kupffer cells and stellate cells by centrifugal elutriation. This method was particularly useful for preparing endothelial cells in high yield (100 X 10(6) cells per liver). Intravenously injected formaldehyde-treated albumin was selectively taken up by the endothelial cells. Isolated endothelial cells in suspension as well as in surface culture maintained their ability to endocytose this ligand.
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PMID:Preparation of isolated liver endothelial cells and Kupffer cells in high yield by means of an enterotoxin. 631 61

Insulin release and beta-cell membrane potentials in response to glucose at 37 and 27 degrees C have been measured simultaneously in single, micro-dissected, perifused islets of Langerhans from normal mice. Insulin release and 45Ca outflow in response to glucose at 37 and 27 degrees C have been measured simultaneously from perfused islets isolated by collagenase digestion from normal rats. The effect of cooling on beta-cell membrane potassium permeability was assessed by changes in measured membrane potential and input resistance (in the mouse) and by changes in 86Rb outflow (in the rat). Resting and active beta-cell membrane parameters (i.e. membrane potential, spike frequency, input resistance, 45Ca outflow and 86Rb outflow), in both mouse and rat islets, were affected only slightly by cooling to 27 degrees C, with temperature coefficients of 2 or lower. At 27 degrees C glucose-stimulated insulin release was inhibited completely in mouse islets and almost completely in rat islets. The temperature coefficients in both preparations were greater than 5. It is concluded that beta-cell electrical activity and changes in membrane permeability induced by glucose are not consequences of insulin release.
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PMID:Cooling dissociates glucose-induced insulin release from electrical activity and cation fluxes in rodent pancreatic islets. 637 Dec 19

The authors report a study in which they evaluate the efficacy of some laboratory parameters for monitoring intrasplenic hepatocyte xenotransplantation (mouse to rat) as an alternative to 99Tc-HIDA dynamic scan and histologic exam. Swiss mouse and wistar rat hepatocytes were obtained with collagenase digestion. Wistar male rats were used as recipient and were allocated into three groups: A) omotransplanted rats; B) xenotransplanted rats; C) xenotransplanted and immunosuppressed (Cyclosporin A: 20 mg/kg/daily orally) rats. All rats underwent > 70% hepatectomy. Blood samples were obtained daily from a femoral vein and AST, ALT, ALP, bilirubin, albumin and urea were measured. No statistical differences were observed among groups and the laboratory parameters tested can't be considered a valid technique to xenotransplant rejection monitoring.
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PMID:[Monitoring of hepatocyte xenotransplantation. Usefulness of various laboratory parameters]. 761 63

In the present study, we examined the roles of the matrix metalloproteinases collagenase, 72-kDa and 92-kDa gelatinase, and proteoglycanase in the tissue remodeling that occurs during luteal development and regression, using a pseudopregnant rat model. Pseudopregnancy was induced in immature female rats by eCG/hCG priming, and animals (n = 3 to 4 per time point) were killed on Days 1, 2, 4, 8, 12, 14, or 16 of pseudopregnancy (Day 0 = time of hCG administration). Ovaries were then removed and analyzed for either matrix metalloproteinase mRNA expression or activity. During luteal development (Day 1 of pseudopregnancy), activity of both collagenase (p = 0.009) and gelatinase (p = 0.0003), but not proteoglycanase (p > or = 0.05), was significantly greater than at all other time points. In accord with gelatinase activity, transcript levels of this enzyme were elevated at Day 1 of pseudopregnancy. Specifically, gelatinase transcript levels for the 72-kDa and 92-kDa enzymes were greatest at Day 1 (p = 0.0003 and p = 0.001, respectively), decreased 3-fold by Day 2,4-fold by Day 4, and reached 10-fold lower levels by Days 8 and 12 of pseudopregnancy. Proteoglycanase transcript could not be detected by Northern analysis during luteal development or any other time point examined in the current study. During the period of luteal maintenance (approximately Days 4-8 of pseudopregnancy in the rat), collagenase and gelatinase displayed basal levels of activity, but only proteoglycanase activity was elevated compared to luteal development levels of this enzyme. During luteal regression (Days 12-14 of pseudopregnancy in the rat), all enzymes displayed basal levels of enzyme activity. In accord with gelatinolytic activity during luteal regression, both 72-kDa and 92-kDa gelatinase mRNA were detectable at baseline levels. In contrast to the baseline levels of collagenolytic activity during luteal regression, collagenase transcript displayed peak values (approximately 8-fold greater than Day 1 levels; p = 0.004) at Day 12 of pseudopregnancy. It is concluded from these studies that collagenase and the gelatinases play a role in the tissue remodeling associated with luteal development, proteoglycanase is associated with luteal maintenance, and collagenase may contribute to the structural regression of the corpus luteum.
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PMID:Collagenase, gelatinase, and proteoglycanase messenger ribonucleic acid expression and activity during luteal development, maintenance, and regression in the pseudopregnant rat ovary. 883 83


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