Characterization and proteolytic activity of a cathepsin L-like polypeptide in endometrium and uterine flushings of cycling, pregnant and steroid-treated ovariectomized gilts

Cathepsin L has been proposed to be involved with the endothelial–chorial type of placentation in the cat. Little information concerning the presence and secretion of cathepsin L is available for a species with noninvasive epitheliochorial placentation such as the pig. Cathepsin L activity in uterine flushings and endometrium from gilts during different days of the oestrous cycle and early pregnancy was analysed through specific substrate metabolism and Western blot analyses with antiserum against cat endometrial cathepsin L. This antiserum was utilized to determine the cellular localization of the enzyme within porcine endometrium. Cathepsin L activity within uterine flushings was elevated on Day 15 of the oestrous cycle and early pregnancy, with activity declining on Day 18. Cat cathepsin L antiserum cross-reacted with a group of 46, 40 and 38 kDa uterine proteins and detected a product within the surface and glandular epithelium of the endometrium. The appearance of the 40 kDa protein was first detected on Day 10 of the oestrous cycle with the 38 kDa proteins appearing on Day 15 and 18 of pregnancy. The 40 and 38 kDa uterine proteins appear to be steroid regulated as 12 days of progesterone administration is necessary to detect the proteins and cathepsin L activity.

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Expression of inter-alpha-trypsin inhibitor heavy chains in endometrium of cyclic and pregnant gilts

Reproduction ◽

10.1530/rep.0.1260621 ◽

2003 ◽

pp. 621-627 ◽

Cited By ~ 15

Author(s):

RD Geisert ◽

MD Ashworth ◽

Keyword(s):

Gene Expression ◽

Extracellular Matrix ◽

Western Blot ◽

Protease Inhibitor ◽

Early Pregnancy ◽

Trypsin Inhibitor ◽

Serine Protease Inhibitor ◽

Oestrous Cycle ◽

Heavy Chains ◽

Formation Of Complexes

Attachment of the placenta to the uterus in pigs involves extracellular interaction between the expanding trophoblastic membrane and the thick glycocalyx present on the uterine epithelial microvilli. Formation of complexes between members of inter-alpha-trypsin inhibitor family may function in the maintenance of the extracellular matrix. This study investigated the change in the inter-alpha-trypsin inhibitor heavy chains (ITIH1, ITIH2, ITIH3 and ITIH4) during the oestrous cycle and early pregnancy in pigs. Gene expression of ITIH1, ITIH2, ITIH3 and ITIH4 was detected in the endometrium of cyclic and pregnant gilts; however, gene expression of ITIH was not altered throughout the oestrous cycle or early pregnancy. Western blot analysis with an ITIH antiserum identified the possible linkage forms of ITIH with the serine protease inhibitor, bikunin. Pregnancy altered the release of the various inter-alpha-inhibitor forms from the endometrium during the period of trophoblastic attachment. The results from this study indicate that the inter-alpha-trypsin inhibitor family plays an important role in maintenance of the uterine surface glycocalyx during placental attachment in pigs.

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Chemerin Affects P4 and E2 Synthesis in the Porcine Endometrium during Early Pregnancy

International Journal of Molecular Sciences ◽

10.3390/ijms23020945 ◽

2022 ◽

Vol 23 (2) ◽

pp. 945

Author(s):

Marlena Gudelska ◽

Kamil Dobrzyn ◽

Marta Kiezun ◽

Katarzyna Kisielewska ◽

Edyta Rytelewska ◽

...

Keyword(s):

Western Blot ◽

Early Pregnancy ◽

Luteal Phase ◽

Oestrous Cycle ◽

Signalling Pathway ◽

Signalling Pathways ◽

Steroidogenic Enzyme ◽

Akt Phosphorylation ◽

Pathway Activation

Chemerin, belonging to the adipokine family, exhibits pleiotropic activity. We hypothesised that the adipokine could be involved in the regulation of steroidogenesis in the porcine endometrium. Thus, the aim of this study was to determine the effect of chemerin on the key steroidogenic enzyme proteins’ abundance (Western blot), as well as on P4 and E2 secretion (radioimmunoassay) by the porcine endometrium during early pregnancy and the mid-luteal phase of the oestrous cycle. Moreover, we investigated the hormone impact on Erk and Akt signalling pathway activation (Western blot). Chemerin stimulated E2 production on days 10 to 11 of pregnancy. On days 10 to 11 and 15 to 16 of gestation, and on days 10 to 11 of the cycle, chemerin enhanced the expression of StAR and all steroidogenic enzyme proteins. On days 12 to 13 of pregnancy, chemerin decreased StAR and most of the steroidogenic enzyme proteins’ abundance, whereas the P450C17 abundance was increased. On days 27 to 28 of pregnancy, chemerin increased StAR and P450C17 protein contents and decreased 3βHSD protein amounts. It was noted that the adipokine inhibited Erk1/2 and stimulated Akt phosphorylation. The obtained results indicate that chemerin affected P4 and E2 synthesis through the Erk1/2 and Akt signalling pathways.

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Effects of interferon-tau and progesterone on oestrogen-stimulated expression of receptors for oestrogen, progesterone and oxytocin in the endometrium of ovariectomized ewes

Reproduction Fertility and Development ◽

10.1071/rd9960843 ◽

1996 ◽

Vol 8 (5) ◽

pp. 843 ◽

Cited By ~ 28

Author(s):

TE Spencer ◽

MA Mirando ◽

JS Mayes ◽

GH Watson ◽

TL Ott ◽

...

Keyword(s):

Early Pregnancy ◽

Oestrous Cycle ◽

Glandular Epithelium ◽

Interferon Tau ◽

Pr Protein ◽

Prostaglandin F2 Alpha ◽

Combined Actions ◽

To Receive ◽

Protein Treatment

The effects of recombinant ovine interferon-tau (IFN-tau) and progesterone on oestrogen-stimulated expression of endometrial receptors for oestrogen (ER), progesterone (PR) and oxytocin (OTR) were determined in ovariectomized ewes. Cyclic ewes (n = 16) were ovariectomized and fitted with uterine catheters on Day 4 of the oestrous cycle (Day O, oestrous) and assigned randomly in 2 x 2-factorial arrangement to receive daily intrauterine injections of either recombinant ovine IFN-tau (roIFN-tau; 2 x 10(7) anti-viral units) or control proteins from Day 11 to Day 15 and 50 mg progesterone from either Day 4 to Day 10 (E-P) or Day 4 to Day 15 (E+P). All ewes received 50 micrograms oestradiol-17 beta on Days 13, 14 and 15 and were hysterectomized on Day 16. In control ewes, endometrial ER mRNA, PR protein and OTR density were greater in E-P- than E+P- treated ewes. In E-P ewes, roIFN-tau decreased oestrogen-stimulated increases in ER and OTR, but not PR expression compared with control ewes. In E+P ewes, endometrial ER mRNA and protein, PR mRNA and protein, and OTR levels were lower in roIFN-tau-treated ewes than control ewes. Immunoreactive ER and PR were absent in the endometrial luminal and superficial glandular epithelium of roIFN-tau compared with control ewes, but were present in the deep glandular epithelium and stroma regardless of steroid or protein treatment. These results indicate that progesterone affects oestrogen-induced increases in endometrial ER, PR and OTR expression in the PR+ deep glandular epithelium and stroma, whereas IFN-tau suppresses oestrogen-induced increases ER, PR and OTR expression in the PR- luminal and superficial glandular epithelium. These combined actions of IFN-tau and progesterone to suppress oestrogen-induced increases in endometrial OTR formation would prevent pulsatile production of luteolytic prostaglandin F2 alpha by the endometrium during early pregnancy.

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Immunocytochemical localization of prostaglandin synthase in the ovine uterus during the oestrous cycle and in early pregnancy

Reproduction Fertility and Development ◽

10.1071/rd9900311 ◽

1990 ◽

Vol 2 (4) ◽

pp. 311 ◽

Cited By ~ 16

Author(s):

LA Salamonsen ◽

JK Findlay

Keyword(s):

Epithelial Cells ◽

Early Pregnancy ◽

Stromal Cells ◽

Cellular Localization ◽

In Vitro Release ◽

Oestrous Cycle ◽

Intense Staining ◽

Endometrial Cells ◽

Ovine Uterus

Prostaglandin (PG) synthase has been localized by immunocytochemistry within the ovine uterus throughout the oestrous cycle and in early pregnancy. On Day 4 of the cycle, PG synthase was located primarily in the stromal cells in caruncular and intercaruncular tissue with little staining in the epithelium. On Days 14 through to 16, the most intense staining was in the luminal epithelial cells (caruncular and intercaruncular) and in epithelial cells of glands close to the uterine lumen. PG synthase was also located in the intercaruncular stromal cells, particularly close to the myometrium. Staining for the enzyme on Day 10 was intermediate between that of Day 4 and Day 14. On Day 15 of pregnancy, the pattern of staining was identical to that on Day 15 of the cycle, with no detectable difference in intensity. When endometrial cells (cycle, Day 14) were cultured with and without ovine trophoblast protein-1 (3 ng mL-1) in vitro, release of PGE and PGF2 alpha was attenuated (54% and 47% of control respectively) but no differences were observed in the intensity of staining for PG synthase in the cells. These results demonstrate marked cyclical changes in the endometrial cell types producing PGs, suggesting differential regulation of PG synthase. In addition, it appears that conceptus-induced changes in PGF2 alpha release do not occur via changes in the concentration or cellular localization of PG synthase, but rather that the activity of the enzyme is modified.

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Changes in oestrogen receptor protein, mRNA expression and localization in the endometrium of cyclic and pregnant gilts

Reproduction Fertility and Development ◽

10.1071/rd9930247 ◽

1993 ◽

Vol 5 (3) ◽

pp. 247 ◽

Cited By ~ 63

Author(s):

RD Geisert ◽

RM Brenner ◽

RJ Moffatt ◽

JP Harney ◽

T Yellin ◽

...

Keyword(s):

Epithelial Cells ◽

Mrna Expression ◽

Oestrogen Receptor ◽

Cellular Localization ◽

Oestrous Cycle ◽

Glandular Epithelium ◽

Receptor Protein ◽

Immunocytochemical Staining ◽

Positive Staining ◽

Intense Staining

Conceptus secretion of oestrogen on Day 11 of gestation is involved with establishment of pregnancy in the pig. Changes in oestrogen receptor (ER) protein, mRNA and cellular localization in the endometrium were evaluated during the oestrous cycle and early pregnancy of the gilt. In nonpregnant gilts, concentration of nuclear ER in the endometrium increased from Days 0 to 12 followed by a decline on Day 15 of the oestrous cycle. In pregnant gilts, changes in endometrial nuclear ER during Days 10, 12, 15 and 18 were similar to that in cyclic pigs. Analysis of endometrial ER mRNA expression did not detect any difference between cyclic and pregnant pigs between Days 10 and 15 postoestrus. Expression of ER mRNA in endometrium of cyclic and pregnant gilts was greatest on Day 10 followed by a decline on Day 15. Endometrial ER mRNA increased on Day 18 of the oestrous cycle, but remained low during pregnancy. Immunocytochemical localization of ER in the endometria of cyclic and pregnant gilts indicated that there was intense staining for ER in stromal cells and moderate to strong staining in surface and glandular epithelial cells during oestrus (Day 0) and Day 18 of the oestrous cycle. However, stromal ER staining was absent from Days 5 to 15 of the oestrous cycle and continued to be suppressed on Day 18 of pregnancy. Immunocytochemical staining of ER in the surface and glandular epithelium was readily detectable from Days 0 to 12 of the oestrous cycle and during pregnancy. Intensity of staining for ER declined in surface epithelial cells on Day 15 in both cyclic and pregnant pigs whereas positive staining for ER in glandular epithelium was absent. Staining for ER on uterine surface epithelial cells increased during pro-estrus (Day 18) of cyclic gilts but remained similar to Day 15 in pregnant gilts. Changes in endometrial ER protein, mRNA and localization in surface epithelium are consistent with a physiological role for conceptus oestrogen secretion in uterine function and maternal recognition of pregnancy in the pig.

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Immunocytochemical localization and changes in endometrial progestin receptor protein during the porcine oestrous cycle and early pregnancy

Reproduction Fertility and Development ◽

10.1071/rd9940749 ◽

1994 ◽

Vol 6 (6) ◽

pp. 749 ◽

Cited By ~ 84

Author(s):

RD Geisert ◽

TN Pratt ◽

FW Bazer ◽

JS Mayes ◽

GH Watson

Keyword(s):

Progesterone Receptor ◽

Early Pregnancy ◽

Cellular Localization ◽

Oestrous Cycle ◽

Receptor Protein ◽

Surface Epithelium ◽

Immunocytochemical Localization ◽

Intense Staining ◽

Pr Protein ◽

Progestin Receptor

Changes of progesterone receptor (PR) protein and cellular localization in the endometrium were evaluated during the oestrous cycle and early pregnancy of the gilt. During the oestrous cycle, the concentration of total PR protein within the endometrium was highest on Days 0-5 and decreased on Day 10. The endometrial concentration of PR reached a nadir on Day 12 and this level was maintained throughout the remainder of the oestrous cycle (Day 18). In pregnant gilts, the concentration of endometrial PR protein from Day 10 to Day 18 was similar to that in cyclic gilts. Western blot analysis with antiserum specific for the A and B isoforms of PR indicated that porcine endometrium expresses both isoforms of PR. Immunostaining was detectable for both the A and B isoforms of PR from Day 0 to Day 12 of the oestrous cycle. However, no staining was observed on Day 15 and Day 18 of the oestrous cycle or pregnancy Immunocytochemical localization of PR in the endometrium of cyclic gilts and pregnant gilts indicated that there was intense staining for PR in surface epithelium and glandular epithelium during oestrus (Day 0) and on Day 5. However, the staining was less intense on Day 7 and Day 10 of the oestrous cycle and no epithelial staining was observed after Day 12. PRs were present in the stroma and myometrium throughout the oestrous cycle and early pregnancy. The presence of conceptuses during pregnancy did not affect the loss of PR from the uterine epithelium after Day 10 of gestation. Down-regulation of epithelial PR might be one factor involved in the timing of luteolysis during the oestrous cycle as well as conceptus growth and placentation during early pregnancy.

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