PMID- 25323036
OWN - NLM
STAT- MEDLINE
DCOM- 20150804
LR  - 20151119
IS  - 1741-7899 (Electronic)
IS  - 1470-1626 (Linking)
VI  - 149
IP  - 1
DP  - 2015 Jan
TI  - Regulation of ovulatory genes in bovine granulosa cells: lessons from siRNA 
      silencing of PTGS2.
PG  - 21-9
LID - 10.1530/REP-14-0337 [doi]
AB  - 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.
CI  - (c) 2015 Society for Reproduction and Fertility.
FAU - Shrestha, Ketan
AU  - Shrestha K
AD  - Department of Animal SciencesThe Robert H. Smith Faculty of Agriculture, Food and 
      Environment, The Hebrew University of Jerusalem, Rehovot 76100, IsraelDepartment 
      of Reproductive Immunology and PathologyInstitute of Animal Reproduction and Food 
      Research, Polish Academy of Science, Olsztyn, PolandDepartment of Reproductive 
      BiologyLeibniz Institute of Farm Animal Biology, Dummerstorf, GermanyDepartment 
      of Ruminant ScienceAgricultural Research Organization, Bet-Dagan, Israel.
FAU - Lukasik, Karolina
AU  - Lukasik K
AD  - Department of Animal SciencesThe Robert H. Smith Faculty of Agriculture, Food and 
      Environment, The Hebrew University of Jerusalem, Rehovot 76100, IsraelDepartment 
      of Reproductive Immunology and PathologyInstitute of Animal Reproduction and Food 
      Research, Polish Academy of Science, Olsztyn, PolandDepartment of Reproductive 
      BiologyLeibniz Institute of Farm Animal Biology, Dummerstorf, GermanyDepartment 
      of Ruminant ScienceAgricultural Research Organization, Bet-Dagan, Israel.
FAU - Baufeld, Anja
AU  - Baufeld A
AD  - Department of Animal SciencesThe Robert H. Smith Faculty of Agriculture, Food and 
      Environment, The Hebrew University of Jerusalem, Rehovot 76100, IsraelDepartment 
      of Reproductive Immunology and PathologyInstitute of Animal Reproduction and Food 
      Research, Polish Academy of Science, Olsztyn, PolandDepartment of Reproductive 
      BiologyLeibniz Institute of Farm Animal Biology, Dummerstorf, GermanyDepartment 
      of Ruminant ScienceAgricultural Research Organization, Bet-Dagan, Israel.
FAU - Vanselow, Jens
AU  - Vanselow J
AD  - Department of Animal SciencesThe Robert H. Smith Faculty of Agriculture, Food and 
      Environment, The Hebrew University of Jerusalem, Rehovot 76100, IsraelDepartment 
      of Reproductive Immunology and PathologyInstitute of Animal Reproduction and Food 
      Research, Polish Academy of Science, Olsztyn, PolandDepartment of Reproductive 
      BiologyLeibniz Institute of Farm Animal Biology, Dummerstorf, GermanyDepartment 
      of Ruminant ScienceAgricultural Research Organization, Bet-Dagan, Israel.
FAU - Moallem, Uzi
AU  - Moallem U
AD  - Department of Animal SciencesThe Robert H. Smith Faculty of Agriculture, Food and 
      Environment, The Hebrew University of Jerusalem, Rehovot 76100, IsraelDepartment 
      of Reproductive Immunology and PathologyInstitute of Animal Reproduction and Food 
      Research, Polish Academy of Science, Olsztyn, PolandDepartment of Reproductive 
      BiologyLeibniz Institute of Farm Animal Biology, Dummerstorf, GermanyDepartment 
      of Ruminant ScienceAgricultural Research Organization, Bet-Dagan, Israel.
FAU - Meidan, Rina
AU  - Meidan R
AD  - Department of Animal SciencesThe Robert H. Smith Faculty of Agriculture, Food and 
      Environment, The Hebrew University of Jerusalem, Rehovot 76100, IsraelDepartment 
      of Reproductive Immunology and PathologyInstitute of Animal Reproduction and Food 
      Research, Polish Academy of Science, Olsztyn, PolandDepartment of Reproductive 
      BiologyLeibniz Institute of Farm Animal Biology, Dummerstorf, GermanyDepartment 
      of Ruminant ScienceAgricultural Research Organization, Bet-Dagan, Israel 
      rina.meidan@mail.huji.ac.il.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20141016
PL  - England
TA  - Reproduction
JT  - Reproduction (Cambridge, England)
JID - 100966036
RN  - 0 (Biomarkers)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Vasodilator Agents)
RN  - 1F7A44V6OU (Colforsin)
RN  - 9002-67-9 (Luteinizing Hormone)
RN  - EC 1.14.99.1 (Cyclooxygenase 2)
RN  - K7Q1JQR04M (Dinoprostone)
SB  - IM
MH  - Animals
MH  - Biomarkers/*metabolism
MH  - Blotting, Western
MH  - Cattle
MH  - Cells, Cultured
MH  - Colforsin/pharmacology
MH  - Cyclooxygenase 2/*chemistry/genetics/*metabolism
MH  - Dinoprostone/pharmacology
MH  - Female
MH  - Gene Expression Regulation/*drug effects
MH  - Granulosa Cells/cytology/drug effects/*metabolism
MH  - Luteinizing Hormone/pharmacology
MH  - Ovulation/*genetics
MH  - RNA, Messenger/genetics
MH  - RNA, Small Interfering/*genetics
MH  - Real-Time Polymerase Chain Reaction
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Vasodilator Agents/pharmacology
EDAT- 2014/10/18 06:00
MHDA- 2015/08/05 06:00
CRDT- 2014/10/18 06:00
PHST- 2014/10/18 06:00 [entrez]
PHST- 2014/10/18 06:00 [pubmed]
PHST- 2015/08/05 06:00 [medline]
AID - REP-14-0337 [pii]
AID - 10.1530/REP-14-0337 [doi]
PST - ppublish
SO  - Reproduction. 2015 Jan;149(1):21-9. doi: 10.1530/REP-14-0337. Epub 2014 Oct 16.