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פותח על ידי קלירמאש פתרונות בע"מ -
Regulation of ovulatory genes in bovine granulosa cells: Lessons from siRNA silencing of PTGS2
Year:
2015
Source of publication :
Reproduction
Authors :
מועלם, עוזי
;
.
Volume :
149
Co-Authors:
Shrestha, K., Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food, and Environment, Hebrew University of Jerusalem, Rehovot, Israel
Lukasik, K., Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland
Baufeld, A., Department of Reproductive Biology, Leibniz Institute of Farm Animal Biology, Dummerstorf, Germany
Vanselow, J., Department of Reproductive Biology, Leibniz Institute of Farm Animal Biology, Dummerstorf, Germany
Moallem, U., Department of Ruminant Science, Agricultural Research Organization, Bet-Dagan, Israel
Meidan, R., Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food, and Environment, Hebrew University of Jerusalem, Rehovot, Israel
Facilitators :
From page:
21
To page:
29
(
Total pages:
9
)
Abstract:
Prostaglandin endoperoxide synthase-2 (PTGS2), tumour necrosis factor-alpha-induced protein-6 (TNFAIP6), pentraxin-3 (PTX3), epidermal growth factor-like factors: amphiregulin (AREG) and epiregulin (EREG) are essential for successful ovulation. In this study, we compared the induction of these ovulatory genes in bovine granulosa cells (GCs) in vivo (after LH surge) and in vitro (forskolin (FRS) treatment). These genes were markedly stimulated in GCs isolated from cows 21 h after LH-surge. In isolated GCs, FRS induced a distinct temporal profile for each gene. Generally, there was a good agreement between the in vivo and in vitro inductions of these genes except for PTX3. Lack of PTX3 induction in isolated GCs culture suggests that other follicular compartments may mediate its induction by LH. Next, to study the role of PTGS2 and prostaglandins (PGs) in the cascade of ovulatory genes, PTGS2 was silenced with siRNA. PTGS2 siRNA caused a marked and specific knockdown of PTGS2 mRNA and PGE2 production (70% compared with scrambled siRNA) in bovine GCs. Importantly, PTGS2 silencing also reduced AREG, EREG and TNFAIP6 mRNA levels but not PTX3. Exogenous PGE2 increased AREG, EREG and TNFAIP6 mRNA levels, further confirming that these genes are prostanoid dependent. A successful and specific knockdown of PTGS2 was also achieved in endometrial cells (EndoCs) expressing PTGS2. Then, cholesterol-conjugated PTGS2 (chol-PTGS2) siRNA that facilitates cells' entry was investigated. In EndoCs, but not in GCs, chol-PTGS2 siRNA succeeded to reduce PTGS2 and PGE2 levels even without transfection reagent. PTGS2 knockdown is a promising tool to critically examine the functions of PTGS2 in the reproductive tract. © 2015 Society for Reproduction and Fertility.
Note:
Related Files :
Animal
Animals
cattle
Female
gene silencing
Genetics
Granulosa Cells
metabolism
RT-PCR (Real-Time Polymerase Chain Reaction)
עוד תגיות
תוכן קשור
More details
DOI :
10.1530/REP-14-0337
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
22954
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:55
You may also be interested in
Scientific Publication
Regulation of ovulatory genes in bovine granulosa cells: Lessons from siRNA silencing of PTGS2
149
Shrestha, K., Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food, and Environment, Hebrew University of Jerusalem, Rehovot, Israel
Lukasik, K., Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland
Baufeld, A., Department of Reproductive Biology, Leibniz Institute of Farm Animal Biology, Dummerstorf, Germany
Vanselow, J., Department of Reproductive Biology, Leibniz Institute of Farm Animal Biology, Dummerstorf, Germany
Moallem, U., Department of Ruminant Science, Agricultural Research Organization, Bet-Dagan, Israel
Meidan, R., Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food, and Environment, Hebrew University of Jerusalem, Rehovot, Israel
Regulation of ovulatory genes in bovine granulosa cells: Lessons from siRNA silencing of PTGS2
Prostaglandin endoperoxide synthase-2 (PTGS2), tumour necrosis factor-alpha-induced protein-6 (TNFAIP6), pentraxin-3 (PTX3), epidermal growth factor-like factors: amphiregulin (AREG) and epiregulin (EREG) are essential for successful ovulation. In this study, we compared the induction of these ovulatory genes in bovine granulosa cells (GCs) in vivo (after LH surge) and in vitro (forskolin (FRS) treatment). These genes were markedly stimulated in GCs isolated from cows 21 h after LH-surge. In isolated GCs, FRS induced a distinct temporal profile for each gene. Generally, there was a good agreement between the in vivo and in vitro inductions of these genes except for PTX3. Lack of PTX3 induction in isolated GCs culture suggests that other follicular compartments may mediate its induction by LH. Next, to study the role of PTGS2 and prostaglandins (PGs) in the cascade of ovulatory genes, PTGS2 was silenced with siRNA. PTGS2 siRNA caused a marked and specific knockdown of PTGS2 mRNA and PGE2 production (70% compared with scrambled siRNA) in bovine GCs. Importantly, PTGS2 silencing also reduced AREG, EREG and TNFAIP6 mRNA levels but not PTX3. Exogenous PGE2 increased AREG, EREG and TNFAIP6 mRNA levels, further confirming that these genes are prostanoid dependent. A successful and specific knockdown of PTGS2 was also achieved in endometrial cells (EndoCs) expressing PTGS2. Then, cholesterol-conjugated PTGS2 (chol-PTGS2) siRNA that facilitates cells' entry was investigated. In EndoCs, but not in GCs, chol-PTGS2 siRNA succeeded to reduce PTGS2 and PGE2 levels even without transfection reagent. PTGS2 knockdown is a promising tool to critically examine the functions of PTGS2 in the reproductive tract. © 2015 Society for Reproduction and Fertility.
Scientific Publication
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