Catechol-O-methyltransferase and methoxyestradiols participate in the intraoviductal nongenomic pathway through which estradiol accelerates egg transport in cycling rats

Alexis Parada-Bustamante, Pedro A. Orihuela, Mariana Ríos, Patricia A. Navarrete-Gómez, Catherina A. Cuevas, Luis A. Velasquez, Manuel J. Villalón, Horacio B. Croxatto

Resultado de la investigación: Article

26 Citas (Scopus)

Resumen

Estradiol (E2) accelerates oviductal egg transport through intraoviductal nongenomic pathways in cyclic rats and through genomic pathways in pregnant rats. This shift in pathways, which we have provisionally designated as intracellular path shifting (IPS), is caused by mating-associated signals and represents a novel and hitherto unrecognized phenomenon. The mechanism underlying IPS is currently under investigation. Using microarray analysis, we identified several genes the expression levels of which changed in the rat oviduct within 6 hours of mating. Among these genes, the mRNA level for the enzyme catechol-O-methyltransferase (COMT), which produces methoxyestradiols from hydroxyestradiols, decreased 6-fold, as confirmed by real-time PCR. O-methylation of 2-hydroxyestradiol was up to 4-fold higher in oviductal protein extracts from cyclic rats than from pregnant rats and was blocked by OR486, which is a selective inhibitor of COMT. The levels in the rat oviduct of mRNA and protein for cytochrome P450 isoforms 1A1 and 1B1, which form hydroxyestradiols, were detected by RT-PCR and Western blotting. We explored whether methoxyestradiols participate in the pathways involved in E 2-accelerated egg transport. Intrabursal application of OR486 prevented E2 from accelerating egg transport in cyclic rats but not in pregnant rats, whereas 2-methoxyestradiol (2ME) and 4-methoxyestradiol mimicked the effect of E2 on egg transport in cyclic rats but not in pregnant rats. The effect of 2ME on egg transport was blocked by intrabursal administration of the protein kinase inhibitor H-89 or the antiestrogen ICI 182780, but not by actinomycin D or OR486. We conclude that in the absence of mating, COMT-mediated formation of methoxyestradiols in the oviduct is essential for the nongenomic pathway through which E2 accelerates egg transport in the rat oviduct. Yet unidentified mating-associated signals, which act directly on oviductal cells, shut down the E2 nongenomic signaling pathway upstream and downstream of methoxyestradiols. These findings highlight a physiological role for methoxyestradiols in the female genital tract, thereby confirming the occurrence of and providing a partial explanation for the mechanism underlying IPS.

Idioma originalEnglish
Páginas (desde-hasta)934-941
Número de páginas8
PublicaciónBiology of Reproduction
Volumen77
N.º6
DOI
EstadoPublished - dic 2007

Huella dactilar

Catechol O-Methyltransferase
Ovum
Estradiol
Oviducts
Messenger RNA
Estrogen Receptor Modulators
Dactinomycin
Microarray Analysis
Protein Kinase Inhibitors
Cytochrome P-450 Enzyme System
Methylation
Real-Time Polymerase Chain Reaction
Protein Isoforms
Proteins
Western Blotting

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology
  • Medicine(all)

Citar esto

Parada-Bustamante, Alexis ; Orihuela, Pedro A. ; Ríos, Mariana ; Navarrete-Gómez, Patricia A. ; Cuevas, Catherina A. ; Velasquez, Luis A. ; Villalón, Manuel J. ; Croxatto, Horacio B. / Catechol-O-methyltransferase and methoxyestradiols participate in the intraoviductal nongenomic pathway through which estradiol accelerates egg transport in cycling rats. En: Biology of Reproduction. 2007 ; Vol. 77, N.º 6. pp. 934-941.
@article{fb70fe51f4e44a8a9c2c171253e1fa0f,
title = "Catechol-O-methyltransferase and methoxyestradiols participate in the intraoviductal nongenomic pathway through which estradiol accelerates egg transport in cycling rats",
abstract = "Estradiol (E2) accelerates oviductal egg transport through intraoviductal nongenomic pathways in cyclic rats and through genomic pathways in pregnant rats. This shift in pathways, which we have provisionally designated as intracellular path shifting (IPS), is caused by mating-associated signals and represents a novel and hitherto unrecognized phenomenon. The mechanism underlying IPS is currently under investigation. Using microarray analysis, we identified several genes the expression levels of which changed in the rat oviduct within 6 hours of mating. Among these genes, the mRNA level for the enzyme catechol-O-methyltransferase (COMT), which produces methoxyestradiols from hydroxyestradiols, decreased 6-fold, as confirmed by real-time PCR. O-methylation of 2-hydroxyestradiol was up to 4-fold higher in oviductal protein extracts from cyclic rats than from pregnant rats and was blocked by OR486, which is a selective inhibitor of COMT. The levels in the rat oviduct of mRNA and protein for cytochrome P450 isoforms 1A1 and 1B1, which form hydroxyestradiols, were detected by RT-PCR and Western blotting. We explored whether methoxyestradiols participate in the pathways involved in E 2-accelerated egg transport. Intrabursal application of OR486 prevented E2 from accelerating egg transport in cyclic rats but not in pregnant rats, whereas 2-methoxyestradiol (2ME) and 4-methoxyestradiol mimicked the effect of E2 on egg transport in cyclic rats but not in pregnant rats. The effect of 2ME on egg transport was blocked by intrabursal administration of the protein kinase inhibitor H-89 or the antiestrogen ICI 182780, but not by actinomycin D or OR486. We conclude that in the absence of mating, COMT-mediated formation of methoxyestradiols in the oviduct is essential for the nongenomic pathway through which E2 accelerates egg transport in the rat oviduct. Yet unidentified mating-associated signals, which act directly on oviductal cells, shut down the E2 nongenomic signaling pathway upstream and downstream of methoxyestradiols. These findings highlight a physiological role for methoxyestradiols in the female genital tract, thereby confirming the occurrence of and providing a partial explanation for the mechanism underlying IPS.",
keywords = "17β-estradiol, Catechol-O-methyltransferase, Estrogen receptor, Methoxyestradiols, Oocyte transport, Oviduct, Ovum pick-up/transport",
author = "Alexis Parada-Bustamante and Orihuela, {Pedro A.} and Mariana R{\'i}os and Navarrete-G{\'o}mez, {Patricia A.} and Cuevas, {Catherina A.} and Velasquez, {Luis A.} and Villal{\'o}n, {Manuel J.} and Croxatto, {Horacio B.}",
year = "2007",
month = "12",
doi = "10.1095/biolreprod.107.061622",
language = "English",
volume = "77",
pages = "934--941",
journal = "Biology of Reproduction",
issn = "0006-3363",
publisher = "Society for the Study of Reproduction",
number = "6",

}

Catechol-O-methyltransferase and methoxyestradiols participate in the intraoviductal nongenomic pathway through which estradiol accelerates egg transport in cycling rats. / Parada-Bustamante, Alexis; Orihuela, Pedro A.; Ríos, Mariana; Navarrete-Gómez, Patricia A.; Cuevas, Catherina A.; Velasquez, Luis A.; Villalón, Manuel J.; Croxatto, Horacio B.

En: Biology of Reproduction, Vol. 77, N.º 6, 12.2007, p. 934-941.

Resultado de la investigación: Article

TY - JOUR

T1 - Catechol-O-methyltransferase and methoxyestradiols participate in the intraoviductal nongenomic pathway through which estradiol accelerates egg transport in cycling rats

AU - Parada-Bustamante, Alexis

AU - Orihuela, Pedro A.

AU - Ríos, Mariana

AU - Navarrete-Gómez, Patricia A.

AU - Cuevas, Catherina A.

AU - Velasquez, Luis A.

AU - Villalón, Manuel J.

AU - Croxatto, Horacio B.

PY - 2007/12

Y1 - 2007/12

N2 - Estradiol (E2) accelerates oviductal egg transport through intraoviductal nongenomic pathways in cyclic rats and through genomic pathways in pregnant rats. This shift in pathways, which we have provisionally designated as intracellular path shifting (IPS), is caused by mating-associated signals and represents a novel and hitherto unrecognized phenomenon. The mechanism underlying IPS is currently under investigation. Using microarray analysis, we identified several genes the expression levels of which changed in the rat oviduct within 6 hours of mating. Among these genes, the mRNA level for the enzyme catechol-O-methyltransferase (COMT), which produces methoxyestradiols from hydroxyestradiols, decreased 6-fold, as confirmed by real-time PCR. O-methylation of 2-hydroxyestradiol was up to 4-fold higher in oviductal protein extracts from cyclic rats than from pregnant rats and was blocked by OR486, which is a selective inhibitor of COMT. The levels in the rat oviduct of mRNA and protein for cytochrome P450 isoforms 1A1 and 1B1, which form hydroxyestradiols, were detected by RT-PCR and Western blotting. We explored whether methoxyestradiols participate in the pathways involved in E 2-accelerated egg transport. Intrabursal application of OR486 prevented E2 from accelerating egg transport in cyclic rats but not in pregnant rats, whereas 2-methoxyestradiol (2ME) and 4-methoxyestradiol mimicked the effect of E2 on egg transport in cyclic rats but not in pregnant rats. The effect of 2ME on egg transport was blocked by intrabursal administration of the protein kinase inhibitor H-89 or the antiestrogen ICI 182780, but not by actinomycin D or OR486. We conclude that in the absence of mating, COMT-mediated formation of methoxyestradiols in the oviduct is essential for the nongenomic pathway through which E2 accelerates egg transport in the rat oviduct. Yet unidentified mating-associated signals, which act directly on oviductal cells, shut down the E2 nongenomic signaling pathway upstream and downstream of methoxyestradiols. These findings highlight a physiological role for methoxyestradiols in the female genital tract, thereby confirming the occurrence of and providing a partial explanation for the mechanism underlying IPS.

AB - Estradiol (E2) accelerates oviductal egg transport through intraoviductal nongenomic pathways in cyclic rats and through genomic pathways in pregnant rats. This shift in pathways, which we have provisionally designated as intracellular path shifting (IPS), is caused by mating-associated signals and represents a novel and hitherto unrecognized phenomenon. The mechanism underlying IPS is currently under investigation. Using microarray analysis, we identified several genes the expression levels of which changed in the rat oviduct within 6 hours of mating. Among these genes, the mRNA level for the enzyme catechol-O-methyltransferase (COMT), which produces methoxyestradiols from hydroxyestradiols, decreased 6-fold, as confirmed by real-time PCR. O-methylation of 2-hydroxyestradiol was up to 4-fold higher in oviductal protein extracts from cyclic rats than from pregnant rats and was blocked by OR486, which is a selective inhibitor of COMT. The levels in the rat oviduct of mRNA and protein for cytochrome P450 isoforms 1A1 and 1B1, which form hydroxyestradiols, were detected by RT-PCR and Western blotting. We explored whether methoxyestradiols participate in the pathways involved in E 2-accelerated egg transport. Intrabursal application of OR486 prevented E2 from accelerating egg transport in cyclic rats but not in pregnant rats, whereas 2-methoxyestradiol (2ME) and 4-methoxyestradiol mimicked the effect of E2 on egg transport in cyclic rats but not in pregnant rats. The effect of 2ME on egg transport was blocked by intrabursal administration of the protein kinase inhibitor H-89 or the antiestrogen ICI 182780, but not by actinomycin D or OR486. We conclude that in the absence of mating, COMT-mediated formation of methoxyestradiols in the oviduct is essential for the nongenomic pathway through which E2 accelerates egg transport in the rat oviduct. Yet unidentified mating-associated signals, which act directly on oviductal cells, shut down the E2 nongenomic signaling pathway upstream and downstream of methoxyestradiols. These findings highlight a physiological role for methoxyestradiols in the female genital tract, thereby confirming the occurrence of and providing a partial explanation for the mechanism underlying IPS.

KW - 17β-estradiol

KW - Catechol-O-methyltransferase

KW - Estrogen receptor

KW - Methoxyestradiols

KW - Oocyte transport

KW - Oviduct

KW - Ovum pick-up/transport

UR - http://www.scopus.com/inward/record.url?scp=36649019924&partnerID=8YFLogxK

U2 - 10.1095/biolreprod.107.061622

DO - 10.1095/biolreprod.107.061622

M3 - Article

C2 - 17699737

AN - SCOPUS:36649019924

VL - 77

SP - 934

EP - 941

JO - Biology of Reproduction

JF - Biology of Reproduction

SN - 0006-3363

IS - 6

ER -