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)

A technique utilizing Pregnant Mare's Serum Gonadotropin and Human Chorionic Gonadotropin treatment of hens (Gallus domesticus), followed by manual ovulation of the excised follicles, was developed to obtain a large number of mature ova. The intact ova were used to test whether acrosin, partially purified from the spermatozoa of the cock (Gallus domesticus), partially purified rabbit testicular acrosin and commercial preparations of several hydrolytic enzymes could dissolve the inner vitelline membrane. Enzymes were applied to pieces of filter paper placed on the ovum. Cock acrosin and endopeptidases such as trypsin, chymotrypsin, collagenase and elastase hydrolyzed the membrane whereas exopeptidases such as leucine aminopeptidase and carboxypeptidase A did not. Phospholipase A, sulfatase, hyaluronidase, beta-glucuronidase and rabbit testicular acrosin also failed to hydrolyze the membrane. Cock acrosin hydrolysis of the ovum surface was inhibited by soybean trypsin inhibitor. The surface of the ovum over the germinal disc region was hydrolyzed more quickly by cock acrosin than the surface over other regions of the ovum. Acrosin from cock sperm caused the release of trichloroacetic acid soluble material absorbing at 280 nm from sonicated preparations of inner vitelline membranes. Hydrolysis was greatest at pH 8.0 and was inhibited by soybean trypsin inhibitor.
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PMID:Hydrolysis of the hen egg vitelline membrane by cock sperm acrosin and other enzymes. 0 Apr 54

Crude preparations of collagenase, which have been used commonly for tissue dissociation, contain proteases that dissolve zonae pellucidae of hamster and mouse oocytes without reducing the ability of the oolemma to fuse with spermatozoa. This gentle proteolytic removal of zona is particularly useful for the study of sperm-oocyte fusion in mice, as trypsin, chymotrypsin and pronase damage the mouse oolemma.
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PMID:Collagenase as an agent for dissolving the zona pellucida of hamster and mouse oocytes. 166 55

Epithelial cells of the rat's epididymal caput were cultivated according to own modification of the Kierszenbaum's method [1981]. The said modification consisted in developing primary cultures of the epithelial cells in the epididymal duct by making use of small tubular segments instead of deisolated cells of the whole epididymal duct wall. Such small segments of the tubules were procured by resorting to mechanical isolation and a 4-grade enzymatic isolation with trypsin and collagenase, whereupon the produced suspension of cells and tubules was filtered through a grid, the meshes of which being 40 X 50 microns in diameter. The cultures were made up exclusively of the tubular segments that had remained on the grid. The utilized technique of isolation gets rid of tubules from the external layer of muscle cells and fibroblasts as well as spermatozoa still prior to the inception of the culture, and provides the possibility to obtain a pure population of epithelial cells. The latter cells have the capacity to migrate from tubular fragments, and to form monolayer cultures. In the conducted cultures the epithelial cells commence secreting PAS-positive substance which was evidenced by means of histochemical and microscope-electron examinations.
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PMID:Modified procedure for isolation of epithelial cells of rat epididymal caput. 280 90

A method for estimating the concentration of spermatozoa in the rat cauda epididymidis is described. Treatment of a sperm suspension with 0.05% collagenase for 20-60 min or 0.025% trypsin for 1-2 min at 34-37 degrees C was found to result in consistently homogeneous sperm. Sperm concentration ranged from 152.5 to 230.0 X 10(7) spermatozoa/ml, with a mean of 187.7 (+/- 5.6 SEM) X 10(7) spermatozoa/ml.
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PMID:A method for estimating the concentration of spermatozoa in the rat cauda epididymidis. 283 33

Trout testes at various stages of maturation were dissociated by perfusion at 12 degrees C with collagenase plus pronase and then with collagenase alone, followed by slight shaking overnight in 1% bovine albumin. This step provided a suspension of isolated somatic and germ cells, clusters of interstitial cells, and either intact spermatogenetic cysts (meiotic testes) or clusters of Sertoli cells (other testes). Most of the spermatozoa were removed from the testis cell suspension by centrifugation in Percoll (density 1.065 g/ml). Sertoli and Leydig cells were prepared by a two-step separation method: 1) the testis cell suspension was separated by sedimentation at unit gravity into "isolated cell" and "cell cluster" populations; 2) these populations were fractionated by isopyknic centrifugation in Percoll gradients. In terms of somatic cell composition, a nearly pure Sertoli cell (clusters) population was obtained between 1.017 and 1.033 g/ml and a Leydig cell (clusters) enriched population of between 1.033 and 1.048 g/ml (testes resuming spermatogenesis) or 1.048 and 1.062 g/ml (other testes). These various cell populations were cultured in modified Leibovitz L15 medium for 10-15 days. When seeded, the Sertoli cells had a normal ultrastructure that remained unchanged for at least 10 days, and the steroidogenic activity of Leydig cells could be stimulated by salmon gonadotropin. Leydig cells remained 3 beta-HSD positive and produced progesterone and 17 alpha, 20 beta-OH progesterone for at least 11 days. This study points out that viable and differentiated trout somatic testicular cells can be prepared and cultured for several days.
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PMID:Trout Sertoli and Leydig cells: isolation, separation, and culture. 323 52

A method for estimating the concentration of spermatozoa in the rat cauda epididymidis is described. Treatment of a sperm suspension with 0.05% collagenase for 20-60 min or 0.025% trypsin for 1-2 min at 34-37 degrees C was found to result in consistently homogeneous sperm. Sperm concentration ranged from 152.5 to 230.0 x 10(7) spermatozoa/ml, with a mean of 187.7 +/- 5.6 (SEM) x 10(7) spermatozoa/ml.
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PMID:Determination of spermatozoa concentration in the rat cauda epididymidis. 345 91

A method for the isolation and culture of epididymal epithelial cells obtained from pubertal and old adult rats is described. This method permits the establishment of primary cultures of these cells in monolayers from aggregates isolated from whole epididymides and major epididymal anatomical segments (caput, corpus, and cauda) after trypsin and collagenase digestions. A large number of cultured epididymal cells retain a differentiated function as demonstrated by the immunocytochemical and radioimmunoassay finding of acidic epididymal glycoprotein, a spermatozoa-coating protein secreted by the principal cells of rat epididymis. The proliferative potential of cultured epididymal cells obtained from pubertal and old adult donors can be documented by [3H]thymidine labeling and mitotic indices without significant loss of gene expression for acidic epididymal glycoprotein. Results of this study demonstrate that epididymal epithelial cells, consisting of a predominant population of principal cells, can be isolated, cultured, and maintained for up to 3 months.
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PMID:Isolation, culture, and immunocytochemical characterization of epididymal epithelial cells from pubertal and adult rats. 701 41

Enzymes and chemicals were used to analyse the biochemical structure of the antigenic epitope recognized by GDA-J/F3 monoclonal antibody (MoAb) in the human sperm tail fibrous sheath. Treatment of sperm dried onto slides with trypsin or dispase enzymes abolished their immunofluorescence staining with GDA-J/F3 MoAb, thus indicating the proteinaceous nature of the antigen. The proteolytic cleavage of GDA-J/F3 protein by trypsin, which also caused sperm decapitation, indicated the presence of peptide bonds involving the carboxyl groups of the basic amino acids, arginine and/or lysine. The epitope was also glycosylated as demonstrated by its sensitivity to sodium metaperiodate treatment which was dose-dependent. The GDA-J/F3 antigenic epitope lacked sialic acid since pre-treatment of spermatozoa with sialidase enzyme (neuraminidase) had no effect on their reactivity with the antibody. The lack of collagenous domains in the GDA-J/F3 antigen was demonstrated by the failure of collagenase to abrogate sperm immunostaining with the MoAb. Furthermore, type VII collagen of the skin basement membrane (BM) was previously thought of as a potential target antigen for GDA-J/F3 MoAb. This was ruled out since several monoclonal and polyclonal antibodies failed to detect the antigen in the spermatozoa using immunofluorescence and Western blotting. These data, therefore, show that the target antigen for GDA-J/F3 MoAb is a non-collagenous asialo-glycoprotein, and by inference provide the first evidence for the glycosylation of the sheath proteins as another step of post-translational modification occurring during sperm tail development.
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PMID:The target antigen for GDA-J/F3 monoclonal antibody in the human sperm tail fibrous sheath is a non-collagenous asialo-glycoprotein: implications and significance. 752 22

AJ-FS9 is one of a new series of monoclonal antibodies (mAbs) raised by immunizing mice with isolated human sperm tail fibrous sheath (FS). Using indirect immunofluorescence (IIF) of human spermatozoa dried onto slides, the AJ-FS9 mAb reacted with the principal piece of occasional spermatozoa. Following their enzymatic treatment with trypsin, dispase or collagenase, but not sulphatase, all the spermatozoa were stained at their principal piece. The ultrastructural localization of the antigens to the FS was established by immunogold electron microscopy, which showed the distribution of gold particles on the FS outer surface of spermatozoa sequentially treated with 1% Triton and dispase; spermatozoa demembranated by Triton alone showed no reaction. For biochemical characterization, spermatozoa were lysed with 1% Triton, and the sperm pellet was run through a reducing sodium dodecyl sulphate-polyacrylamide gel electrophoresis, Western blotted and immunostained with AJ-FS9. The results showed the reaction of the antibody with three protein bands with molecular masses ranging between 46 and 56 kDa. IIF screening of human testicular cryostat sections with AJ-FS9 mAb showed its reactivity with occasional sperm tails; but following their dispase treatment, all spermatozoa were stained. The restricted staining of the assembled FS of maturing sperm tails indicated the late appearance of the antigens during spermatogenesis. The antibody did not react with sperm cell precursors or other cell types within/without the seminiferous tubules. Untreated and dispase-treated frozen sections of skin, oesophagus, tongue, liver, kidney, stomach, ileum or their blood vessels showed no reaction.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:AJ-FS9 monoclonal antibody detects masked antigens within the human sperm tail fibrous sheath. 784 12

The aim of the present study was to isolate and characterize digestive gland cells of the bivalve mollusc Mytilus galloprovincialis Lmk. The digestive gland of bivalves is a complex organ composed of digestive and connective tissues but it is also invaded by the reproductive tissue as gametogenesis proceeds. The digestive tissue is comprised of stomach and intestinal epithelial cells, ciliated and non-ciliated duct cells, digestive cells and basophilic cells. the last two cell types are found lining the epithelium of the blind-ending digestive tubules and are the main focus of this study. Two different approaches were assayed for cell isolation, i.e., explant culture techniques and mechanical plus enzymatical digestion techniques. Cell viability was tested by trypan blue exclusion, neutral red uptake and ultrastructural analysis. The explant cultures were often contaminated with bacteria and spermatozoa and, moreover, cells migrating out of the explants possibly corresponded to hemocytes and not to digestive tissue cells. Mechanical plus enzymatical digestion with collagenase was concluded to be the method of choice for digestive cell isolation, with a percentage of about 30% of isolated cells corresponding to digestive cells. The ultrastructure of digestive cells isolated with the latter procedure closely resembled that of mussel digestive cells in vivo.
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PMID:Isolation and morphofunctional characterization of mussel digestive gland cells in vitro. 912 36


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