PEPTIDYL PEPTIDE HYDROLASES, CATHEPSIN AND COLLAGENASE (?) OF THE RAT UTERUS DURING POSTPARTUM INVOLUTION

1971 ◽  
Vol 67 (3) ◽  
pp. 563-569 ◽  
Author(s):  
Dieter Platt ◽  
Hanke Frackenpohl
Keyword(s):  
Protein Content ◽  
Dna Content ◽  
Post Partum ◽  
Collagenase Activity ◽  
Rat Uterus ◽  
Cathepsin Activity ◽  
Postpartum Involution ◽  
Peptide Hydrolases

ABSTRACT 1. The activity of cathepsin and of collagenase in the rat uterus during parturition and in post partum involution increased significantly 60 h after parturition. 2. Cathepsin activity was measured using haemoglobin as substrate at pH 4.1. 3. Collagenase activity was tested with 4-phenyl-azo-benzyloxy-carbonyl-L-prolyl-L-leucyl-glycyl-L-prolyl-D-arginine-OH as substrate at pH 8.5. 4. The protein content of the rat uterus remained constant during post partum involution. 5. The DNA content increased significantly.

scholarly journals A latent form of collagenase in the involuting rat uterus and its activation by a serine proteinase

1977 ◽  
Vol 161 (3) ◽  
pp. 535-542 ◽  
Author(s):  
J F Woessner
Keyword(s):  
Wide Spectrum ◽  
Post Partum ◽  
Active Form ◽  
Serine Proteinase ◽  
Apparent Molecular Weight ◽  
Lima Bean ◽  
Trypsin Inhibitors ◽  
Collagenase Activity ◽  
Rat Uterus ◽  
Latent Form

1. The involuting rat uterus displays an extremely rapid breakdown of collagen. Collagenase activity can be assayed directly in the insoluble 6000g pellet of uterine homogenates. At 1 day post partum, about 85% of this collagenase activity is in a latent form. 2. This latent form can be activated by trypsin or by a serine proteinase present in the uterine pellets. 3. The activating enzyme of the tissue is inhibited by a wide spectrum of trypsin inhibitors, including Trasylol, soya-bean and lima-bean trypsin inhibitors, snail inhibitor and di-isopropyl phosphoro-fluoridate. Partial inhibition is produced by benzamidine, phenylmethanesulphonyl fluoride, epsilon-aminohexanoate, leupeptin, antipain and alpha1-antitrypsin. Ovomucoid, 7-amino-1-chloro-3-tosylamido-1-heptan-2-one and 1-chloro-4-phenyl-3-(N-benzyloxy-carbonyl)amino-L-butan-2-one are not inhibitory. 4. Extraction of uterine pellets with 0.1 M-CaCl2 at 60 degrees C releases both latent and active collagenase. Exclusion chromatography on Sephadex G-100 gives an apparent molecular weight of approx. 77000 for the latent form and 66000 for the active form. The latent form is suggested to be a zymogen of collagenase.

scholarly journals Total, latent and active collagenase during the course of post-partum involution of the rat uterus. Effect of oestradiol

1979 ◽  
Vol 180 (1) ◽  
pp. 95-102 ◽  
Author(s):  
J F Woessner
Keyword(s):  
Post Partum ◽  
Serine Proteinase ◽  
Temporal Correlation ◽  
Total Activity ◽  
Insoluble Fraction ◽  
Collagenase Activity ◽  
Rat Uterus ◽  
Latent Form ◽  
Tissue Homogenates

1. Total, active and latent collagenase activities were determined by direct assay of tissue homogenates. 2. The rate of collagen breakdown during post-partum involution of the rat uterus is correlated with the total activity of collagenase. Both are low at parturition, reach a maximum within 24h and fall slowly to low values of 5 days post partum. This temporal correlation strongly supports the hypothesis that collagenase participates in collagen breakdown in vivo. 3. Further support for this hypothesis is provided by the finding that oestradiol-17 beta (100 micrograms/day, intraperitoneally injected), which inhibits the breakdown of collagen by 36% during the first 4 days of involution, produces a closely corresponding decrease in total collagenase activity. 3. The effect of oestradiol in lowering collagenase activity is not due to alterations in collagen substrate, collagenase kinetic behaviour or latent-to-active enzyme conversion. 4. Of the total assayable collagenase, about 35% is fully active and 65% is in a latent form. 5. About 70% of this latent form can be activated by a serine proteinase found, together with collagenase, in the insoluble fraction of uterine homogenates.

scholarly journals Oestradiol inhibits collagen breakdown in the involuting rat uterus

1972 ◽  
Vol 127 (4) ◽  
pp. 705-713 ◽  
Author(s):  
Janet N. Ryan ◽  
J. Frederick Woessner
Keyword(s):  
Cathepsin D ◽  
Density Gradient Centrifugation ◽  
Post Partum ◽  
Gradient Centrifugation ◽  
Specific Radioactivity ◽  
Rat Uterus ◽  
Oestradiol Treatment ◽  
Cathepsin D Activity ◽  
Stimulation Of

1. The earlier observation (Woessner, 1969) of oestradiol inhibition of collagen breakdown is confirmed and extended. Administration of 100μg of oestradiol-17β/day to parturient rats strongly inhibits the loss of collagen from the involuting uterus. Three experiments show that this effect is due to an inhibition of collagen degradation rather than to a stimulation of collagen synthesis. 2. Uterine collagen was labelled with hydroxy[14C]-proline by the administration of [14C]proline near the end of pregnancy. By 3 days post partum, control uteri lost 83% of their collagen and 90% of their hydroxy[14C]proline. Uteri from oestradiol-treated rats lost only 50% of both total and labelled hydroxyproline, with no decrease in the specific radioactivity of the hydroxyproline. 3. Incorporation of [14C]proline into uterine collagen hydroxyproline in vivo was not affected by oestradiol treatment. 4. Urinary excretion of hydroxyproline was increased in post-partum control rats and decreased in oestradiol-treated rats. 5. An enzyme capable of cleaving 4-phenylazobenzyloxycarbonyl-l-prolyl-l-leucylglycyl- l-prolyl-d-arginine (a substrate for clostridial collagenase) increased in activity in the post-partum uterus and was unaffected by oestradiol treatment. 6. Uterine homogenates digested uterine collagen extensively at pH3.2. This digestion was unaffected by the oestradiol treatment. 7. Lysosomal fractions prepared by density-gradient centrifugation of uterine homogenates contained coincident peaks of cathepsin D activity and peptide-bound hydroxyproline. The cathepsin D and hydroxyproline contents of this peak were unaffected by oestradiol treatment.

IL-1 beta and IL-6 induce hyperplasia and hypertrophy of cultured guinea pig airway smooth muscle cells

1995 ◽  
Vol 78 (4) ◽  
pp. 1555-1563 ◽  
Author(s):  
S. De ◽  
E. T. Zelazny ◽  
J. F. Souhrada ◽  
M. Souhrada
Keyword(s):  
Smooth Muscle ◽  
Guinea Pig ◽  
Protein Content ◽  
Airway Smooth Muscle ◽  
Dna Content ◽  
Thymidine Incorporation ◽  
Interleukin 1 ◽  
Total Protein Content ◽  
Leucine Incorporation ◽  
Airway Smooth Muscle Cells

Guinea pig airway smooth muscle (ASM) cells were maintained in a primary tissue culture (passages 1–3). Cells were exposed to human recombinant interleukin-1 beta (IL-1 beta; 20–100 pg/ml) or interleukin-6 (IL-6; 1–4 ng/ml) in the presence of indomethacin (1 microgram/ml) for up to 5 days. Proliferation of ASM cells was assessed with two techniques, direct counting of cells with a hemacytometer and [3H]thymidine incorporation corrected for total protein content. Hypertrophy of ASM cells was assessed by [3H]leucine incorporation (evaluation of protein synthesis), determination of total DNA content, DNA content per cell, and protein content per cell. We observed that the exposure of ASM cells to human recombinant IL-1 beta or IL-6, in all studied concentrations, significantly increased the number of cells as well as [3H]thymidine incorporation into ASM cells. We also found that exposure of ASM to these two cytokines increased [3H]leucine incorporation into the ASM cells and increased protein content and DNA content per single cell. These changes were also concentration dependent. We conclude that the two proinflammatory cytokines, IL-1 beta and IL-6, which are present in asthmatic lungs, increased the proliferation of ASM cells (hyperplasia) as well as their overall size and size of their nuclei, as measured by biochemical markers. These findings are compatible with the presence of ASM hypertrophy.

Changes in protein during development of Triturus embryos

Development ◽  
1973 ◽  
Vol 30 (3) ◽  
pp. 647-659
Author(s):  
Hiroshi Imoh ◽  
Tsutomu Minamidani
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Sodium Carbonate ◽  
Dna Content ◽  
Total Protein ◽  
Soluble Protein ◽  
Nuclear Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tail Bud ◽  
Stage Of Development

The present paper reports basic data on DNA content, protein content, and protein synthesis in Triturus pyrrhogaster embryos during development from cleavage to the hatching stage. Except for measurements of DNA and total protein contents, embryos were labeled with sodium carbonate-14C for 10 h and fractionated into embryonic cell components, i.e. cytoplasmic mass, yolk and pigment granules, and nuclei, in a discontinuous density gradient of sucrose. The protein content and the radioactivity incorporated into protein were measured in each fraction. Those fractions combining protein soluble in buffer at pH 8·3 and in 0·25 N-HCl were further studied with polyacrylamide gel electrophoresis. In the newt embryo, four stages of active DNA increase were observed when cultured at constant temperature; they were gastrula, neurula, late tail-bud, and before-hatching stages. Total protein per embryo decreased from 3 to 2 mg during the development studied. The content of cytoplasmic soluble protein per embryo was low and constant throughout development. Synthesis of the fraction was observed at the earliest stage of development studied though the rate was not high and specific activity of the soluble protein increased during development. Qualitative changes in the newly synthesized protein were observed. With the yolk fraction, synthesis of protein, other than from probable contamination with the cytoplasmic fraction, was not detected and a detailed description was omitted. Changes were observed at two stages of development in the synthesis of nuclear protein soluble in buffer at pH 8·3, the first at gastrulation and the second at late tail-bud stage. The change at gastrulation seemed to be the start of syntheses of the nuclear soluble proteins, while quantitative enhancement rather than qualitative change was noticed at late tail-bud stage. Most of the nuclear protein soluble in 0·25 N-HCI was histone. The histone content increased in accordance with increase in the DNA content and the rate of DNA accumulation was accompanied by proportionate incorporation of radioactivity into histone. Among histone fractions, unique behaviour of the very lysine-rich histone was observed. The availability of [14C]sodium carbonate in rough estimations of protein synthesis in embryos and significance of the data obtained have been discussed.

The Role of Neurogenic Bioamines in Nerve Fibers of Uterus during the Postpartum Involution in Experimental Animal Models

2021 ◽  
Vol 27 ◽  
Author(s):  
Sergey V. Dindyaev ◽  
Narasimha M Beeraka ◽  
Denis V. Kasatkin ◽  
Elizaveta V. Mikhaylenko ◽  
Siva G. Somasundaram ◽  
...  
Keyword(s):  
Glyoxylic Acid ◽  
Sympathetic Nerves ◽  
Biological Action ◽  
Nerve Fibers ◽  
Structural Elements ◽  
Rat Uterus ◽  
Postpartum Involution ◽  
Tissue Specimens ◽  
Nulliparous Female ◽  
Uterine Body

Background: Biogenic amines (BAs) secreted by the sympathetic neural apparatus of rat uterus is reported to be conducive to the uterine functional activity during postpartum involution; the imbalance in BAs ratio could confer postpartum reproductive disorders including improper postpartum involution. Objective: The changes in density of uterine sympathetic nerves implicated in the pathology of endometriosis, adenomyosis, and delayed uterine involution. The present study is aimed to ascertain ‘serotonin’ and ‘catecholamine’ concentrations in mesenteric mast cells (MCs), and structural elements of nerve fibers across the perivascular plexuses (PPs) and single sympathetic nerve terminals (SST). Methods: Furthermore, the density of their spatial distribution (SDP and SDT) in the uterine body, cervix, and mesometrium was determined during postpartum involution. Tissue specimens of postpartum uterus were obtained from 55 nulliparous female Wistar outbred strain rats, which were grouped according to the days after parturition at the time of sacrifice. The nerve fibers of PP and SST exhibited emerald green fluorescence, which was detected by glyoxylic acid fluorescence technique; the fluorescence invoked by BAs was identified by microspectrofluorimetry. Results: Concentrations of BAs were extensive in the varicosities of PP and SST on the 10th day. However, the highest BA concentrations were found in structural elements of PP in the uterine mesometrium in the initial days of postpartum. In mesenteric MC, serotonin and catecholamines were at the highest concentration on 10th day postpartum. Histamines peaked on the 6th day. Conclusion: SDP and SDT were increased significantly in all structural elements of uterine nerve fibers in the uterine body and cervix compared to SDP in mesentery. Considering that catecholamines and serotonin are antagonists in many aspects of their biological action, the ratio of BAs should be well-balanced to maintain anabolic-catabolic equilibrium in the rat uterus.

Distribution of amylase and chymotrypsin in four regions of the rat pancreas

1981 ◽  
Vol 59 (8) ◽  
pp. 884-886 ◽  
Author(s):  
Adrien R. Beaudoin ◽  
Pierre St-Jean ◽  
Ginette Girard
Keyword(s):  
Protein Content ◽  
Acinar Cell ◽  
Dna Content ◽  
Pancreatic Tissue ◽  
Weight Basis ◽  
Secretory Proteins ◽  
Fresh Weight ◽  
Fresh Weight Basis ◽  
Rat Pancreas

Recent observations have confirmed the existence of more than one pool of secretory proteins in the rat pancreatic tissue. To determine if these different pools could be located in the different regions of the rat pancreas, the amylase and chymotrypsin contents have been measured in the biliary, duodenal, gastric, and splenic regions. On a tissue DNA content, protein content, or a fresh weight basis, the proportions of these two enzymes are comparable in the four regions. It is therefore postulated that heterogeneity of enzyme composition exists either within these regions or within the acinar cell itself.

EFFECT OF PROGESTERONE ON COLLAGEN BREAKDOWN AND TISSUE COLLAGENOLYTIC ACTIVITY IN THE INVOLUTING RAT UTERUS

1975 ◽  
Vol 66 (3) ◽  
pp. 357-362 ◽  
Author(s):  
JOUKO HALME ◽  
J. F. WOESSNER
Keyword(s):  
Post Partum ◽  
Collagenolytic Activity ◽  
Rat Uterus ◽  
Uterine Involution ◽  
Wet Weight

SUMMARY Progestational agents were studied for their effects on collagenolytic activity and loss of collagen and wet weight from the involuting post-partum rat uterus. Administration of very large doses of progesterone (80–150 mg/day) significantly retarded uterine involution and loss of collagen. This was accompanied by a significant reduction in uterine collagenolytic activity. By 72 h post partum, uteri of rats treated with 150 mg progesterone/day had wet weights 30% above, collagen 85% above, and collagenolytic activity 45% below, those of the control uteri. Similar effects were produced by 17α-acetoxy-6α-methylprogesterone at the same dosage levels. However, the progestational agent 6α-chloro-17α-acetoxypregna-4,6-diene-3,20dione acetate had no effect in this system.

EFFECT OF OESTRADIOL ON THE POST-PARTUM RAT UTERUS: PEROXIDASE ACTIVITY AND COLLAGEN BREAKDOWN

1980 ◽  
Vol 85 (3) ◽  
pp. 387-391 ◽  
Author(s):  
J. F. WOESSNER ◽  
JANET N. RYAN
Keyword(s):  
Epithelial Cells ◽  
Peroxidase Activity ◽  
Post Partum ◽  
Cell Type ◽  
Rat Uterus ◽  
Uterine Epithelial Cells ◽  
Uterine Involution ◽  
Tenfold Increase

Treatment of parturient rats with 100 μg oestradiol/day caused a significant retardation of uterine involution and collagen breakdown. The post-partum uterus had a peroxidase activity of 180 μmol H2O2 reduced/min per uterus. Treatment with oestradiol caused an eight- to tenfold increase in this activity within 3–4 days. Treatment of rats with 4 mg cortisol/day commencing 3 days prepartum had no effect on uterine peroxidase activity but it blocked the oestradiol-induced increase in peroxidase. Cortisol had no effect on collagen breakdown and did not reverse the inhibition of collagen breakdown produced by oestradiol. It is postulated that the effects of oestradiol on peroxidase activity are mediated by uterine epithelial cells but that oestradiol effects on collagen breakdown may be mediated by another cell type.

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