Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo

P. D. Burns; G. A. Graf; S. H. Hayes; W. J. Silvia

doi:10.1016/S0739-7240(00)00080-1

Effect of oxytocin on expression of cytosolic phospholipase A₂ mRNA and protein in ovine endometrial tissue in vivo

P. D. Burns, G. A. Graf, S. H. Hayes, W. J. Silvia

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

The induction of endometrial prostaglandin (PG) F_2α synthesis by oxytocin is dependent upon activation of phospholipase (PL) A₂ and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF_2α synthesis by inducing synthesis of cytosolic PLA₂ (cPLA₂). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA₂ protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA₂ protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA₂ mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF_2α synthesis by activating cPLA₂ protein that is already present in an inactive form.

Original language	English
Pages (from-to)	237-246
Number of pages	10
Journal	Domestic Animal Endocrinology
Volume	19
Issue number	4
DOIs	https://doi.org/10.1016/S0739-7240(00)00080-1
State	Published - 2000

Bibliographical note

Funding Information:
The authors wish to thank Doug Yelton and Brent Broaddus for their extensive efforts in these experiments and to Dr. Phillip Chapman for helping with the statistical analyses. This research was supported in part by grants from the Kentucky Agricultural Station and USDA (96-35203-3495) to P.D.B. This report is published with the approval of the Kentucky Agricultural Experiment Station (Publication No. 99-07-125).

ASJC Scopus subject areas

Food Animals
Animal Science and Zoology
Endocrinology

Access to Document

10.1016/S0739-7240(00)00080-1

Cite this

@article{b026806045c0495494981e8b5a109167,

title = "Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo",

abstract = "The induction of endometrial prostaglandin (PG) F2α synthesis by oxytocin is dependent upon activation of phospholipase (PL) A2 and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF2α synthesis by inducing synthesis of cytosolic PLA2 (cPLA2). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F2α (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA2 protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA2 protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA2 mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF2α synthesis by activating cPLA2 protein that is already present in an inactive form.",

author = "Burns, {P. D.} and Graf, {G. A.} and Hayes, {S. H.} and Silvia, {W. J.}",

note = "Funding Information: The authors wish to thank Doug Yelton and Brent Broaddus for their extensive efforts in these experiments and to Dr. Phillip Chapman for helping with the statistical analyses. This research was supported in part by grants from the Kentucky Agricultural Station and USDA (96-35203-3495) to P.D.B. This report is published with the approval of the Kentucky Agricultural Experiment Station (Publication No. 99-07-125).",

year = "2000",

doi = "10.1016/S0739-7240(00)00080-1",

language = "English",

volume = "19",

pages = "237--246",

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T1 - Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo

AU - Burns, P. D.

AU - Graf, G. A.

AU - Hayes, S. H.

AU - Silvia, W. J.

N1 - Funding Information: The authors wish to thank Doug Yelton and Brent Broaddus for their extensive efforts in these experiments and to Dr. Phillip Chapman for helping with the statistical analyses. This research was supported in part by grants from the Kentucky Agricultural Station and USDA (96-35203-3495) to P.D.B. This report is published with the approval of the Kentucky Agricultural Experiment Station (Publication No. 99-07-125).

PY - 2000

Y1 - 2000

N2 - The induction of endometrial prostaglandin (PG) F2α synthesis by oxytocin is dependent upon activation of phospholipase (PL) A2 and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF2α synthesis by inducing synthesis of cytosolic PLA2 (cPLA2). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F2α (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA2 protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA2 protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA2 mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF2α synthesis by activating cPLA2 protein that is already present in an inactive form.

AB - The induction of endometrial prostaglandin (PG) F2α synthesis by oxytocin is dependent upon activation of phospholipase (PL) A2 and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF2α synthesis by inducing synthesis of cytosolic PLA2 (cPLA2). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F2α (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA2 protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA2 protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA2 mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF2α synthesis by activating cPLA2 protein that is already present in an inactive form.

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