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)

The effects of different neurotransmitters were tested in vitro on a hypothalamic tissue, collagenase-digested isolated anterior pituitary and Leydig cell suspension system by measuring the testosterone production of the Leydig cells. Neurotransmitters were used in concentrations of 0.25, 1.0, 2.5, 5.0, and 10.0 micrograms/ml incubation medium. Dopamine in doses of 1.0, 2.5, and 5.0 micrograms/ml increased the hypothalamic tissue-induced pituitary-testis activation, while it had no direct effect on pituitary and Leydig cells. Noradrenaline in the concentration range 2.5--10.0 micrograms/ml decreased the luteinizing-hormone-releasing-hormone (LHRH) sensitivity of the pituitary cells. 5.0 and 10.0 micrograms/ml 5-hydroxytryptamine decreased the testosterone production and the hCG sensitivity of the isolated Leydig cells. Carbamylcholine and pilocarpine had no action on the in vitro system at the different levels studied.
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PMID:Study of the effects of neurotransmitters on the hypothalamus-pituitary-testis function in in vitro cell suspension system. 4 66

To determine whether specific hormonal responses were involved in the production of cryoprotectant (glucose) by liver of the freeze tolerant wood frog, Rana sylvatica, metabolically active hepatocytes were isolated in reasonable yields (mean 20.1 +/- 1.30% SEM, n = 29) by in situ liver perfusion with collagenase. Freshly isolated cells from autumn-collected frogs contained large amounts of glycogen (650 mumol glucosyl units/g packed cells) and produced glucose from this endogenous reserve at a rate of 10 mumol g-1 hr-1 at 0 degrees. Glucose output from cells was highly responsive to the addition of hormones; rates of glucose release increased 2.1-, 1.7-, and 1.7-fold with the addition of 10(-7) M bovine glucagon, 10(-7) M epinephrine, and 5 x 10(-6) M dibutyryl-cyclic AMP, respectively. Norepinephrine, 5-hydroxytryptamine, and bovine insulin were without effect at 0.1 microM/l. Hormone stimulation of glucose release was correlated with an increase in both the total activity and the percentage a of glycogen phosphorylase in hepatocytes. However, none of the hormones tested affected the kinetic properties of hepatocyte pyruvate kinase, suggesting the absence of covalent modification control of the enzyme. The data indicate that the freezing-stimulated production of large quantities of glucose as a cryoprotectant by R. sylvatica liver does not involve qualitative differences in the hormonal control of liver glycogenolysis, compared with other lower vertebrates. However, quantitative differences were seen, such as the much greater phosphorylase activity, 4.38 +/- 0.33 mumol min-1 g-1 packed cells, in freshly isolated R. sylvatica hepatocytes compared with 0.36 +/- 0.06 mumol min-1 g-1 in Rana pipiens hepatocytes.
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PMID:Hormonal effects on glycogen metabolism in isolated hepatocytes of a freeze-tolerant frog. 162 97

The ability of alpha 1a- and alpha 1b-adrenergic receptor subtypes to stimulate [3H]inositol phosphate [( 3H]InsP) formation was examined in collagenase-dispersed hepatocytes and renal cells. alpha 1-Adrenergic receptor binding sites were labeled with 125I-BE 2254, and the proportion of alpha 1a and alpha 1b subtypes was determined with chloroethylclonidine (CEC) and WB 4101. Hepatocytes contained only alpha 1b-adrenergic receptors, whereas renal cells had approximately equal proportions of both subtypes. Pretreatment of renal cells with CEC selectively inactivated the alpha 1b subtype, leaving a homogeneous population of alpha 1a receptors. Norepinephrine stimulated [3H]InsP accumulation to a similar extent in both hepatocytes and renal cells. Pretreatment with CEC inactivated this response completely in hepatocytes but only partially in renal cells. WB 4101 was 1000-fold more potent in inhibiting the [3H]InsP response in renal cells than hepatocytes; however, some of this difference was due to rapid metabolism of WB 4101 by hepatocytes. After correction for metabolism, WB 4101 was still 11-fold more potent in inhibiting norepinephrine-stimulated [3H]InsP formation in hepatocytes (alpha 1b) than in CEC-pretreated renal cells (alpha 1a). These results demonstrate that both alpha 1a- and alpha 1b-adrenergic receptor subtypes activate formation of [3H]InsP, although the molecular mechanisms by which these responses occur remain to be determined.
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PMID:Alpha 1-adrenergic receptor subtypes and formation of inositol phosphates in dispersed hepatocytes and renal cells. 216 16

Dispersed cells from the submandibular gland of the male rat were prepared by collagenase treatment to study the mechanism by which immunoreactive tonin is secreted in vitro. Norepinephrine, epinephrine, and phenylephrine stimulated tonin release, an effect that was inhibited by phentolamine but not by propranolol, whereas isoproterenol, carbachol, histamine, and serotonin did not stimulate tonin release. The stimulatory effect elicited by alpha-adrenergic agonists was inhibited by both removal of Ca2+ from the medium and addition of diltiazem and nifedipine, both selective calcium channel blockers. The divalent cation ionophore A23187 stimulated tonin release in the presence of Ca2+, but not in the presence of Mg2+. Dibutyryl cyclic adenosine 3',5'-monophosphate, methylisobutylxanthine, angiotensin II, and vasoactive intestinal peptide had no effect on tonin release. The apparent molecular size of immunoreactive tonin released into the medium under basal and norepinephrine-stimulated conditions was similar to that of standard tonin by gel exclusion chromatography. These data suggest that the in vitro secretion of immunoreactive tonin from rat submandibular gland is initiated by activation of alpha-adrenergic receptors and apparently involves a mechanism dependent not on cyclic adenosine 3',5'-monophosphate, but on the influx of extracellular Ca2+.
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PMID:In vitro secretion of immunoreactive tonin from dispersed rat submandibular gland cells. 301 87

To locate the sites of dopamine (D) production in rat renal cortex, we separated glomeruli and proximal tubules by sieving or centrifugation in Percoll after collagenase digestion. After centrifugation layer I contained 60-80% glomeruli and 20-40% tubule fragments, half of which did not stain with alkaline phosphatase, layer II contained 0-5% glomeruli, 10-25% tubule fragments other than proximal tubules, and 70-85% proximal tubule fragments. Layer IV contained 85-95% proximal tubules. Gluconeogenic rates were (micromoles per hour per gram wet weight) as follows: I, 4 +/- 1; II, 7 +/- 2; and IV, 16 +/- 1. Norepinephrine (NE) content was (picomoles per gram wet weight) I, 310 +/- 30; II, 540 +/- 40; IV, 195 +/- 60. D content was (picomoles per gram wet weight) I, 26 +/- 6; II, 46 +/- 13; IV, 33 +/- 7. Surgical denervation 4-6 days previously reduced the norepinephrine content of layers I and II to 35 +/- 10 (p less than 0.001) and of IV to 60 +/- 20 (p less than 0.05) and the D content of layers I and II to 13 +/- 6 and 6 +/- 6 pmol/g, respectively (p less than 0.01); D content of layer IV was unchanged. D production from 10(-7) M 3,4-dihydroxyphenylalanine (DOPA) was (nanomoles per gram per minute) I, 0.2 +/- 0.03; II, 0.7 +/- 0.1; IV, 1.0 +/- 0.04. DOPA consumption was (nanomoles per gram per minute) I, 0.6 +/- 0.1; II, 1.4 +/- 0.3; and IV, 1.8 +/- 0.2. Denervation did not change D production or DOPA consumption.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Dopamine production by isolated glomeruli and tubules from rat kidneys. 398 98

1. Tubule fragments were isolated by collagenase treatment of guinea pig kidney cortex. 2. 3':5'-Cyclic AMP increased gluconeogenesis from lactate, pyruvate, malate and fructose. 3. Noradrenaline decreased gluconeogenesis from lactate, pyruvate, 2-oxoglutarate and fructose. 4. Angiotensin II slightly, but significantly, increased gluconeogenesis from lactate. 5. Gluconeogenesis from glycerol as sole substrate was negligible. Gluconeogenesis from combinations of glycerol together with either lactate, pyruvate, 2-oxoglutarate or malate was appreciably greater than the sum of the glucose output observed when these substrates were added singly.
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PMID:Gluconeogenesis in guinea pig renal tubule fragments--effects of noradrenaline, 3':5' cyclic AMP and angiotensin II. 613 91

Viable dispersed cell preparations of rat submandibular gland were obtained by a tissue-dissociation procedure using collagenase and gentle mechanical force. The cells released kallikrein-like esterase in a time- and calcium-dependent manner in response to noradrenaline (10 microM) at 30 degrees C. The net loss of kallikrein-like esterase content from the dispersed cells corresponded with the increase in kallikrein-like esterase activity in the suspending medium at all concentrations of noradrenaline. These results indicate the viability and functional integrity of this dispersed cell preparation of rat submandibular gland. alpha-Adrenoceptor agonists such as noradrenaline stimulated kallikrein-like esterase and tonin release in a dose-dependent manner, whereas the beta-adrenoceptor agonist isoprenaline and cholinoceptor agonist methacoline were both inactive. Noradrenaline-induced release of kallikrein-like esterase and tonin were completely blocked by prior addition of the alpha-adrenoceptor antagonist, phenoxybenzamine. It is concluded that the secretion of kallikrein-like esterase and tonin in rat submandibular gland is mediated only via stimulation of alpha-adrenoceptors.
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PMID:Release of kallikrein-like esterase and tonin from dispersed cells of the rat submandibular gland. 614 66

The maximum phosphodiesterase activity (Vmax) with low and high Km was, respectively, 10- and 3-times greater in tissue fragments than in collagenase-isolated adipocytes obtained from subcutaneous fat layers in man. The exposure of such tissue fragments to collagenase of various origins in order to isolate the fat cells resulted in a 60-70% inhibition of phosphodiesterase (PDE) activity. Noradrenaline- and isopropyl noradrenaline-induced rates of lipolysis were more rapid in the isolated fat cells than in the tissue fragments. The sensitivity to catecholamines, however, was the same for the two tissue preparations. Nor did they differ in respect to the effect of theophylline, a PDE inhibitor, on the rate of lipolysis. The time curve for cyclic AMP accumulation was significantly higher in the isolated adipocytes than in tissue fragments in the presence of isopropyl noradrenaline. It is concluded that the greater lipolytic response of collagenase-isolated adipocytes than of tissue fragments to catecholamines may be attributed, at least in some measure, to the higher concentration of cyclic AMP resulting from a decrease in PDE activity.
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PMID:Influence of adipocyte isolation by collagenase on phosphodiesterase activity and lipolysis in man. 624 9

The regulation of secretion of catecholamines from bovine adrenal medulla cells was investigated by use of an improved and highly efficient method for isolating viable and responsive cells from this tissue. The method involves an in situ collagenase perfusion affecting only the connective tissue matrix of the medulla while leaving the cortex intact. The cells released both epinephrine and norepinephrine in response to stimulation by 100 microM acetylcholine. The ratio of epinephrine to norepinephrine in the medium following non-stimulated (basal) release, was similar to that found in the intact cells. On the other hand, a lower ratio of epinephrine to norepinephrine was found in the medium following stimulation by acetylcholine due mainly to preferential secretion of norepinephrine. This release ceased after 15 min of incubation and consisted of 15--20% of the catecholamines initially present in the cells. Exogenous epinephrine was found to inhibit total catecholamine secretion; however, it stimulated norepinephrine release. Addition of isoproterenol caused a stimulation of release while propranolol was inhibitory. Norepinephrine inhibited total release not favoring any specific catechol. Other alpha-agonists, such as clonidine, also had an inhibitory effect. These results suggest a receptor-mediated mechanism for the fine regulation of secretion from the adrenal medulla.
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PMID:Alpha- and beta-receptor control of catecholamine secretion from isolated adrenal medulla cells. 629 65

Effects of adrenergic and cholinergic drugs and prostaglandin E1 on cyclic nucleotide accumulation and parameters of growth and basement membrane synthesis were examined in corneal epithelial cell cultures. 8-bromo-cGMP significantly (p less than 0.05) enhanced incorporation of labeled thymidine and leucine, as did acetylcholine and carbamylcholine, which elevated cGMP and decreased cAMP/cGMP ratio. Responses to acetylcholine were abolished by atropine and alpha-bungarotoxin. Precursor incorporation was inhibited by dibutyryl cAMP and adenosine 5'-monophosphate and by norepinephrine, epinephrine, prostaglandin E1, and theophylline, which significantly elevated cAMP levels and cAMP/cGMP ratio. Propranolol, but not phenoxybenzamine, blocked responses to effective concentrations of norepinephrine. Norepinephrine, PGE1, and dibutyryl cAMP also significantly elevated uptake of labeled glucosamine and incorporation of labeled proline into collagenase-sensitive protein or the hydroxyproline fraction of protein hydrolysates, while acetylcholine had no effect on parameters of basement membrane synthesis. Propranolol blocked responses to norepinephrine. Results were consistent with a cGMP-mediated stimulatory role of the cholinergic transmitter in corneal epithelial growth regulation, cAMP-mediated beta-adrenergic suppression of regrowth and increased basement membrane production after initial injury to the corneal epithelium, and potentiation of the adrenergic effect by prostaglandins.
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PMID:Cholinergic, adrenergic, and PGE1 effects on cyclic nucleotides and growth in cultured corneal epithelium. 629 65


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