Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.36 (hyaluronidase)
 4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Follicles of 28 day-old pregnant mare serum gonadotropin (PMSG)-primed rats, were cultured for up to 24 hours in the presence or absence of ovine gonadotropins, highly purified rat gonadotropins, dibutyryl cyclic AMP (dbcAMP), methylisobutylxanthine (MIX), choleratoxin (CT), or prostaglandin E2 (PGE2). The morphology of the cumulus-oocyte complexes isolated from these follicles was subsequently examined with the light microscope. Cumulus mucification was studied under the different culture conditions using scanning electron microscopy (SEM) and the hyaluronidase sensitivity test. The features of the cumulus-oocyte complexes in the control cultures did not change throughout the incubation period, while complexes from follicles incubated with LH, FSH, dbcAMP, MIX, CT, or PGE2 changed their appearance and accumulated extracellular mucoid material. Treatment of these cumuli with hyaluronidase resulted in lysis of the extracellular mucus and dispersal of the cumulus masses. The results of this study agree with our earlier observation that the maturation of the cumulus-oophorus, which occurs in vivo following the LH surge, can be induced in vitro by either gonadotropins or cAMP. Prostaglandin E2 did not affect cumulus cells, unless incubated enclosed by their follicles. This suggests that this hormone may influence the cumulus cells indirectly, probably via other components.
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PMID:Effect of gonadotropins and prostaglandin on cumulus mucification in cultures of intact follicles. 618 Jan 23

The aim of this study was to establish a monolayer culture system for human ovarian thecal cells and to investigate their morphological and functional characteristics. Theca layers were isolated and digested with collagenase-hyaluronidase solution, and dispersed thecal cells were cultured for 10 days in plastic dishes. Histological examination indicated that there were no contaminating granulosa cells in isolated theca layers. Various histochemical studies revealed abundant lipid droplets and 3 beta-hydroxysteroid dehydrogenase activity in cultured cells. The major steroids secreted were delta 4-androstenedione (delta 4) and progesterone(P). delta 4 secretion was very high during the first 2 days (31.6 +/- 1.9 ng/1 X 10(5) cells/2 days) and declined thereafter. P was secreted in moderate amounts throughout the 10 day culture period (9.0-21.3 ng/1 X 10(5) cells/2 days), while estradiol secretion was very low. Subsequently, the responsiveness of cultured thecal cells to gonadotropins and dibutyryl cyclic AMP (Bu2cAMP) was investigated. LH/HCG and Bu2cAMP stimulated delta 4 and P secretion in a dose-related manner. The maximal effective doses of LH and HCG were both 10 ng/ml, and that of Bu2cAMP was 10(-3)M. In conclusion, it was evident that these monolayer-cultured human thecal cells could maintain their morpho-functional characteristics during culture. Therefore, this culture system will provide an excellent model for further studies on the functional properties of thecal cells.
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PMID:[Monolayer culture of human ovarian thecal cells. A study on morphological and functional characteristics]. 658 14

Deposition of glycosaminoglycan is one of the histological features of Graves' ophthalmopathy. Although retroocular tissue fibroblasts are considered to be responsible for glycosaminoglycan accumulation, it is not known what is stimulating the fibroblasts. There are studies which are in support of and against the role of anti-TSH receptor antibodies in the pathogenesis of Graves' ophthalmopathy. TSH-receptor antibodies increase cAMP as a second messenger in thyroid cells. We studied the effects of dibutyryl cyclic AMP (Bt2 cAMP) on glycosaminoglycan synthesis by retroocular tissue fibroblasts in order to known whether cAMP can modulate glycosaminoglycan synthesis. Retroocular tissue fibroblasts mainly synthesize hyaluronan, the large chondroitin sulfate proteoglycan and the small chondroitin sulfate proteoglycan as glycosaminoglycan in cell culture. The amount of hyaluronan synthesis was measured as [3H]glucosamine incorporation into macromolecule susceptible to hyaluronidase digestion (from Streptomyces hyaluronlyticus). The amount of proteoglycan synthesis was measured as [35S]sulfate incorporation into macromolecules in medium and cell layer fraction. Proteoglycans in medium were further separated into the large proteoglycan and the small proteoglycan on a Superose 6 column. Bt2 cAMP increased both hyaluronan and proteoglycan synthesis by retroocular tissue fibroblasts, especially stimulating the secretion of the large proteoglycan synthesis by retroocular tissue fibroblasts, especially stimulating the secretion of the large proteoglycan. Effects of Bt2 cAMP on glycosaminoglycan synthesis were then compared with those in adult skin fibroblasts. Although the magnitude of response between the two was indistinct, the stimulation of the large proteoglycan synthesis by Bt2 cAMP was more prominent in retroocular tissue fibroblasts. The results suggest that the regulation of glycosaminoglycan synthesis by retroocular tissue fibroblasts is different from that by adult skin fibroblasts. Although further studies are required to determine its actual role, cAMP stimulates glycosaminoglycan synthesis by retroocular tissue fibroblasts and underlies the mechanism in Graves' ophthalmopathy.
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PMID:Effects of dibutyryl cyclic AMP on hyaluronan and proteoglycan synthesis by retroocular tissue fibroblasts in culture. 770 88

1. Prostaglandin E2 (PGE2) is an autacoid that decreases proteoglycan synthesis, increases metalloprotease production by cultured chondrocytes, and can modulate some of the actions of interleukin-1 on cartilage. The objective of the present study was to characterize the subtype of prostaglandin E2 receptor present in bovine chondrocytes in culture. 2. Primary cultures of articular chondrocytes were prepared from slices of bovine carpal cartilage by sequential digestion with type III hyaluronidase, trypsin, type II collagenase, followed by overnight incubation in Dulbecco's Modified Eagle's Medium (DMEM) with type II collagenase, washing, and seeding at a density of 2 x 10(5) cells cm-2 in DMEM with 10% foetal bovine serum. 3. PGE2 and carbaprostacyclin induced dose-dependent increases in intracellular cyclic AMP in bovine chondrocytes in culture. The potencies of these compounds were different, and maximal doses of PGE2 and carbaprostacyclin had an additive effect. PGD2 induced a small increase in intracellular cyclic AMP only at a high concentration (10(-5) M). 4. PGE2 was more potent that the EP2 agonist 11-deoxy-PGE1 at inducing increases in intracellular cyclic AMP. The EP2 agonist butaprost, however, induced only a small increase at a concentration of 10(-5)M. 17-Phenyl-PGE2 (EP1 agonist), sulprostone and MB 28767 (15S-hydroxy-9-oxo-16-phenoxy-omega-tetranorprost-13E-enoic acid) (EP3 agonists) did not induce an increase in intracellular cyclic AMP at concentrations up to 10(-5)M. 5. The EP4 antagonist AH 23848B ([1 alpha(Z),2 beta, 5 alpha]-(+/-) -7-[5-[[(1,1'-biphenyl)-4-yl]methoxyl-2-(4-morpholinyl) -3-oxocyclopentyl]-5-heptenoic acid) antagonized PGE2 but not carbaprostacyclin effects on intracellular cyclic AMP. The Schild plot slope was different from 1 but this could be due to an interaction of PGE2 with IP receptors in high doses. The exact nature of the antagonism by compound AH 23848B could not be definitely established in these experimental conditions. 6. Neither PGE2 nor any of its analogues inhibited the increase in intracellular cyclic AMP induced by forskolin, and pertussis toxin did not alter the response to PGE2, suggesting that no Gi-coupled PGE2 receptors are present in these cells. Stimulation with PGE2 did not induce significant increases in intracellular inositol-trisphosphate levels nor increases in intracellular free calcium as determined by confocal microscopy, suggesting the absence of phospholipase-C-coupled or of calcium channel-coupled PGE2 receptors in bovine chondrocytes in these experimental conditions. 7. These results show for the first time that bovine chondrocytes in culture present a functional PGE2 receptor that has some pharmacological characteristics of an EP4 subtype, as well as an IP receptor.
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PMID:Characterization of the PGE2 receptor subtype in bovine chondrocytes in culture. 884 20

The objective was to compare culture media for in vitro maturation of equine oocytes and for in vitro culture of zygotes produced from IVF of partially zona-removed oocytes. Cumulus-oocyte complexes from slaughterhouse-derived ovaries were washed in m-Dulbecco's PBS and cultured in TCM-199, F10-DMEM or c-F10-DMEM (50% F10-DMEM + 50% F10-DMEM conditioned medium from culture of an equine trophoblast monolayer for 3 or 4 days). All media included FSH, LH, E2, and 10% FCS. After 28 to 30 h maturation, cumulus expansion was scored from 0 (no expansion) to 4 (fully expanded). Oocytes with a 1st polar body were selected for manipulation after removing cumulus cells using hyaluronidase. About one-third of the zona pellucida was cut using a fragment of a razor blade. For fertilization, fresh stallion semen was washed twice in BGM3 (a modified Tyrode's medium) and capacitated with 0.5 mM c-AMP for 3.5 h and 100 microM ionomycin for 15 min and added to oocytes in fert-TALP at 10(6) spermatozoa/mL. After 20 h, some presumptive zygotes were stained, and the rest were cultured in 100% TCM-DMEM conditioned medium. Cumulus expansion in F10-DMEM and c-F10-DMEM was higher (P<0.05) than the TCM-199 control (3.2, 3.5 vs 1.3, on a scale of 0 to 4). However, polar body formation rates were not different among treatments (47, 52 and 50%). The fertilization rates of equine oocytes matured in TCM-199, F10-DMEM and c-F10-DMEM determined by fixing and staining were 41, 35 and 29%, with no significant differences. There were no significant differences among treatments in cleavage rates (36 to 40%), development to morula (3 to 10%), or blastocyst stages (3 to 5%). On Day 14 of culture in c-F10-DMEM treatment, one blastocyst had more than 500 nuclei, but no capsule was formed. In a further study, cleavage rates (46 to 50%) and development to morula (5 to 10%) and blastocyst stages (3 to 8%) were not different (P>0.1) between TCM-DMEM and 100% conditioned TCM-DMEM for culturing embryos. Six embryos (2 morulae and 4 blastocysts) were nonsurgically transferred to 4 recipient mares, but no pregnancy continued.
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PMID:Developmental capacity of equine oocytes matured and cultured in equine trophoblast-conditioned media. 1148 Jun 24


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