scholarly journals Temporal expression and cellular localization of a gastrin-releasing peptide-related gene in ovine uterus during the oestrous cycle and pregnancy

1998 ◽  
Vol 157 (1) ◽  
pp. 139-148 ◽  
Author(s):  
JC Whitley ◽  
A Shulkes ◽  
LA Salamonsen ◽  
D Vogiagis ◽  
M Familari ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cellular Localization ◽  
Major Product ◽  
Oestrous Cycle ◽  
Endometrial Tissue ◽  
Cycle Duration ◽  
Gastrin Releasing Peptide ◽  
Stromal Tissue ◽  
Blastocyst Implantation ◽  
Ovine Uterus

Synthesis of both mRNA and peptide for gastrin-releasing peptide (GRP) has been demonstrated in the pregnant endometrium of sheep and women. However, it is not known whether GRP is synthesized in the sheep uterus during the oestrous cycle. Furthermore the cellular site of GRP mRNA synthesis in the uterus has not been determined. Therefore we examined the synthesis of GRP and determined the cellular location of GRP peptide and mRNA in sheep uterus taken at different times during the oestrous cycle (duration 17 days) and pregnancy (duration 145 days). Northern blot analysis of RNA isolated from ovine endometrium revealed low or no GRP mRNA at days 4, 10, 12 and 14 of the oestrous cycle and a 24-fold rise in GRP mRNA (normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA) between days 14 and 16. A similar pattern was observed during early pregnancy, with a 12-fold rise in GRP mRNA:GAPDH mRNA between days 17 and 20 of pregnancy. Levels of GRP peptide were determined by RIA and found to be low in endometrium isolated at days 4, 10, 12 and 14 of the oestrous cycle (1.0-1.6 pmol/g) and 4 to 5-fold higher at day 16. In situ hybridization localized GRP synthesis to the epithelial cells of the uterine glands at day 16 of the oestrous cycle and at days 17, 20, 40 and 50 of pregnancy. At day 140 of pregnancy diffuse hybridization to cells of the myometrium was also observed. Immunohistochemistry localized GRP peptide to the apical cytoplasm of uterine glandular epithelial cells at day 16 of the oestrous cycle. For samples obtained at day 20 of pregnancy, the area surrounding the glands also showed moderately strong staining. Further staining in the glandular lumen and the stromal tissue surrounding the glands was apparent at day 140 of pregnancy. No GRP immunoreactivity could be detected in the peripheral plasma during the oestrous cycle or the first 20 days of pregnancy. Sizing chromatography of GRP immunoreactivity extracted from endometrial tissue taken at day 10 of the oestrous cycle revealed two peaks that co-eluted with GRP(1-27) and GRP(18-27). However, during luteolysis and oestrus the major peak of GRP immunoreactivity extracted from endometrial tissue was larger than GRP(1-27) and similar to that seen previously in the gravid ovine endometrium. These studies demonstrate that a peptide similar to, but larger than, GRP is a major product of the glandular epithelium of the ovine uterus during the luteal regression phase of the oestrous cycle and post-blastocyst implantation in pregnancy and provide further evidence that GRP-related peptides have important regulatory roles in uterine function.

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

1990 ◽  
Vol 2 (4) ◽  
pp. 311 ◽  
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.

Localization of neutral endopeptidase in the ovine uterus and conceptus during the oestrous cycle and early pregnancy

1995 ◽  
Vol 7 (1) ◽  
pp. 27 ◽  
Author(s):  
SC Riley ◽  
E Wong ◽  
JK Findlay ◽  
LA Salamonsen
Keyword(s):  
Smooth Muscle ◽  
Epithelial Cells ◽  
Smooth Muscle Cells ◽  
Early Pregnancy ◽  
Stromal Cells ◽  
Immunohistochemical Staining ◽  
Muscle Cells ◽  
Neutral Endopeptidase ◽  
Oestrous Cycle ◽  
Ovine Uterus

Neutral endopeptidase (NEP; EC 3.4.24.11), an enzyme which metabolizes several peptides (including oxytocin and endothelins) implicated in the control of uterine function, was found to be localized in the ovine uterus throughout the oestrous cycle and in the uterus and conceptus during early pregnancy, using immunohistochemical techniques. Positive NEP immunoreactivity was found in the endometrium principally in stromal cells, in the vasculature in endothelial and vascular smooth muscle cells, and also weakly in some glandular epithelial cells. In a layer of stromal fibroblasts several cells in thickness underlying the luminal epithelium, staining was much weaker than that in the deeper stromal cells throughout the period examined. NEP staining was also present in smooth muscle cells of the myometrium at all times, and was most intense in the layer of cells adjacent to the endometrium. In the conceptus, NEP immunohistochemical staining was found in uninucleate cells, but not in binucleate trophoblast cells, in epithelial cells of the allantois and amnion, and in the heart and brain of the Day-20 embryo. In ovariectomized ewes treated with oestrogen or progesterone separately or remaining untreated, immunohistochemical staining of NEP was stronger when compared with intact ewes, in caruncular and intercaruncular stroma and epithelia, in glands, in the vasculature and in myometrium. The staining was less intense in all cell types in ewes receiving oestrogen plus progesterone. The expression of NEP and its specific regionalization within the uterus indicate a mechanism by which the availability of biologically important peptides involved in the regulation of the oestrous cycle and implantation, including oxytocin and endothelin, can be controlled by regulation of their metabolism.

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

1993 ◽  
Vol 5 (3) ◽  
pp. 247 ◽  
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.

NEW DATA CONCERNING THE ROLE PLAYED BY PROGESTERONE IN THE CONTROL OF FOLLICULAR GROWTH IN THE RAT

1974 ◽  
Vol 75 (3) ◽  
pp. 569-578 ◽  
Author(s):  
G. Buffler ◽  
S. Roser
Keyword(s):  
Wistar Rats ◽  
Venous Blood ◽  
Oestrous Cycle ◽  
Female Rats ◽  
Cycle Duration ◽  
Follicular Growth ◽  
The Third ◽  
Female Wistar Rats

ABSTRACT The mechanisms involved in the prolongation of the oestrous cycle following LH administration were studied in 4-day cyclic female Wistar rats. In females injected with LH on the morning of dioestrus I there was an increase in ovarian venous blood progesterone as compared with non-injected animals. In both LH-treated females, and those injected with progesterone on the morning of dioestrus I, a slowing up in follicular growth was observed from the afternoon of dioestrus I. The size of follicles greater than 400 urn present in LH or progesterone injected animals on the third day of cycle was similar to the size reached by the same range of follicles in non-injected animals on the second day of the cycle. Hence, the increase in endogenous ovarian progesterone elicited by LH was considered as the cause of the slowing up of follicular growth and therefore of the lengthening of the oestrous cycle duration in female rats injected with LH at the beginning of 4-day cycle.

EFFECT OF ERGOCORNINE METHANESULPHONATE ON THE LUTEOTROPHIC ACTIVITY OF THE ECTOPIC MOUSE TROPHOBLAST

1968 ◽  
Vol 59 (3) ◽  
pp. 442-446 ◽  
Author(s):  
G. H. Zeilmaker
Keyword(s):  
Direct Effect ◽  
Oestrous Cycle ◽  
Kidney Capsule ◽  
Blastocyst Implantation

ABSTRACT Virgin mice received 2 blastocysts under the kidney capsule. Following an oestrous cycle which occurred 1–4 days after the operation, nearly all the animals became pseudopregnant. It was found that pseudopregnancy induced by ectopic trophoblast could not be terminated by ergocornine administration. In contrast, this drug was effective in interrupting pseudopregnancy induced by sterile mating. The results confirm previous findings a) that placental luteotrophic hormone starts acting soon after blastocyst implantation and b) that ergocornine exerts no direct effect on the ovary.

Oestrous cycle and gestation length in the musky rat-kangaroo, Hypsiprymnodon moschatus (Potoroidae : Marsupialia)

2001 ◽  
Vol 49 (1) ◽  
pp. 37 ◽  
Author(s):  
Shan Lloyd
Keyword(s):  
Epithelial Cells ◽  
Captive Breeding ◽  
Common Ancestor ◽  
Post Partum ◽  
Reproductive Physiology ◽  
Oestrous Cycle ◽  
Gestation Length ◽  
Vaginal Smears ◽  
Breeding Colony ◽  
Male And Female

Wild-caught male and female H. moschatus were maintained in a captive breeding colony. Vaginal smears were taken three times a week until oestrous cycles were detected and gestation lengths approximated. Thereafter, smears were usually taken daily when oestrus was expected. The gestation period (considered to be the number of days from the detection of sperm in the smear until the day young were found in the pouch) was found to last 19 days. Sperm were usually detected in the smear two days before the influx of semi-cornified and cornified epithelial cells, which occurred 17 days before parturition. A pre- or post-partum oestrus was not detected and females did not return to oestrus until at least 6 days after the removal of the last pouch young. H. moschatus has the shortest recorded gestation for any macropod, and gestation occupies approximately 75% of the oestrous cycle. The reproductive physiology of H. moschatus is similar to that of most phalangerids, which may be indicative of a common ancestor.

Changes in the fine structure of the apical plasma membrane of endometrial epithelial cells during implantation in the rat

1982 ◽  
Vol 55 (1) ◽  
pp. 1-12
Author(s):  
C.R. Murphy ◽  
J.G. Swift ◽  
T.M. Mukherjee ◽  
A.W. Rogers
Keyword(s):  
Plasma Membrane ◽  
Fine Structure ◽  
Epithelial Cells ◽  
Normal Pregnancy ◽  
Ovariectomized Rats ◽  
Apical Plasma Membrane ◽  
Embryonic Cells ◽  
Blastocyst Implantation ◽  
First Contact ◽  
Endometrial Epithelial Cells

In previous work we have shown that ovarian hormones, when injected into ovariectomized rats, alter the fine structure of the plasma membrane of endometrial epithelial cells. In this paper freeze-fractures have been used to study the apical plasma membrane of endometrial epithelial cells of rats during the period of blastocyst implantation of normal pregnancy. On day 1 of pregnancy there were 2354 +/− 114 intramembranous particles (IMPs) per micrometer2 of membrane. The particles were spherical and randomly distributed. On day 5 of pregnancy IMP density rose to 2899 +/− 289 per micrometer2 and some rod-shaped particles were also visible. By day 6 of pregnancy IMP density had risen to 4014 +/− 206 per micrometer2 and there were more rod-shaped IMPs than before. In addition, on day 6 IMPs were also present as rows of particles and some gap-junction-like arrays of particles were also seen. Our findings indicate that there are fine-structural alterations in the apical plasma membrane of endometrial epithelial cells, the site of first contact between maternal and embryonic cells, during the period of early pregnancy. The findings are discussed in the light of suggested mechanisms of blastocyst attachment to the uterine epithelium at implantation.

Spatial Transcriptomics analysis of uterine gene expression in enhancer of Zeste homolog 2 (Ezh2) conditional knockout mice

2021 ◽  
Author(s):  
Ana M Mesa ◽  
Jiude Mao ◽  
Theresa I Medrano ◽  
Nathan J Bivens ◽  
Alexander Jurkevich ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Gene Expression Profiles ◽  
Reproductive Organs ◽  
Conditional Knockout ◽  
Estrogen Signaling ◽  
Uterine Epithelial Cell ◽  
Stromal Tissue

Abstract Histone proteins undergo various modifications that alter chromatin structure, including addition of methyl groups. Enhancer of homolog 2 (EZH2), is a histone methyltransferase that methylates lysine residue 27, and thereby, suppresses gene expression. EZH2 plays integral role in the uterus and other reproductive organs. We have previously shown that conditional deletion of uterine EZH2 results in increased proliferation of luminal and glandular epithelial cells, and RNAseq analyses reveal several uterine transcriptomic changes in Ezh2 conditional (c) knockout (KO) mice that can affect estrogen signaling pathways. To pinpoint the origin of such gene expression changes, we used the recently developed spatial transcriptomics (ST) method with the hypotheses that Ezh2cKO mice would predominantly demonstrate changes in epithelial cells and/or ablation of this gene would disrupt normal epithelial/stromal gene expression patterns. Uteri were collected from ovariectomized adult WT and Ezh2cKO mice and analyzed by ST. Asb4, Cxcl14, Dio2, and Igfbp5 were increased, Sult1d1, Mt3, and Lcn2 were reduced in Ezh2cKO uterine epithelium vs. WT epithelium. For Ezh2cKO uterine stroma, differentially expressed key hub genes included Cald1, Fbln1, Myh11, Acta2, and Tagln. Conditional loss of uterine Ezh2 also appears to shift the balance of gene expression profiles in epithelial vs. stromal tissue toward uterine epithelial cell and gland development and proliferation, consistent with uterine gland hyperplasia in these mice. Current findings provide further insight into how EZH2 may selectively affect uterine epithelial and stromal compartments. Additionally, these transcriptome data might provide the mechanistic understanding and valuable biomarkers for human endometrial disorders with epigenetic underpinnings.

PLASMA AND PITUITARY LUTEINIZING HORMONE CONCENTRATIONS AND PERIPHERAL PLASMA OESTRADIOL CONCENTRATION DURING EARLY PREGNANCY AND AFTER THE ADMINISTRATION OF PROGESTATIONAL STEROIDS IN THE RAT

1972 ◽  
Vol 53 (1) ◽  
pp. 31-35 ◽  
Author(s):  
K. BROWN-GRANT ◽  
C. S. CORKER ◽  
F. NAFTOLIN
Keyword(s):  
Luteinizing Hormone ◽  
Marked Decrease ◽  
Single Injection ◽  
Normal Pregnancy ◽  
Oestrous Cycle ◽  
Pregnant Rats ◽  
Pregnant Rat ◽  
Peripheral Plasma ◽  
Blastocyst Implantation ◽  
Plasma Oestradiol

SUMMARY Plasma luteinizing hormone (LH) concentrations were already lower on Day 2 of pregnancy than at the same time after the preceding ovulation in the non-pregnant rat, and fell progressively up to Day 16 of pregnancy. No evidence was obtained of any increase at the time when the ovulatory surge of LH would have occurred if the animal had not become pregnant. Pituitary LH concentration was lower in mated rats on the morning of Day 0 of pregnancy than in unmated controls on the morning of the day of oestrus. Subsequently it increased slowly to reach a level higher than at any stage of the oestrous cycle by Day 8 of pregnancy and remained high until at least Day 16 of pregnancy. Peripheral plasma oestradiol concentration increased late on Day 2 of pregnancy and was still raised on Day 4 but was never more than about one fourth of the peak concentration seen on the morning of prooestrus during the oestrous cycle. There were similar changes in plasma LH and oestradiol concentrations in the 48 h after a single injection of 2·5mg progesterone on the morning of the day of dioestrus, a procedure that delays ovulation by 1 or 2 days. Administration of a synthetic progestational compound (medroxyprogesterone acetate) to pregnant rats delayed blastocyst implantation and the delay was associated with a marked decrease in peripheral plasma LH to levels below those of normal pregnancy.

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