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The biologic activity of 5��-reduced pregnanes in the late gestation mare

Mares have an atypical hormone profile during pregnancy. Systemic
progesterone (P4) levels approach zero by day 220 of gestation. Other reduced
pregnanes such as 5��-pregnane-3, 20-dione (5��), 5��-pregnane-3��, 20��-diol (����),
3��-hydroxy-5��-pregnan-3-one (3��) and 20��-hydroxy-5��-prenan-3-one (20��),
increase to near ��g/mL levels in the peripheral system of the mare until directly
before parturition when they decrease. This unusual hormone profile during
gestation indicates the possibility that other pregnanes, not P4, are responsible for
uterine quiescence and gonadotropin inhibition during pregnancy. Three
experiments were conducted to determine if these steroids have biologic activity.
Experiment 1 consisted of jugular vein blood samples taken from mares from ten
days pre-partum until the foal heat ovulation, approximately 15 days postpartum.
Samples were analyzed for luteinizing hormone (LH), follicle stimulating hormone
(FSH), and pregnane content. Concentrations of these hormones were analyzed for
serial correlations. There was a serial negative correlation with pregnanes and FSH
(p=0.0138), which were analyzed on a same day basis, day -5 to day of foaling.
There also was a positive correlation with pregnanes and FSH analyzed from day of
foaling to 10 days post-foaling (p<0.00l). There was also a significant negative
correlation (p=0.0196) between pregnanes and LH, analyzed on a lag basis, day -5
to day of foaling for pregnanes, and day -5 to day of ovulation for LH. There was
also a significant negative correlation when pregnanes were analyzed from day of
foaling to 10 days post foaling, and LH was analyzed from 10 days before
ovulation to day of ovulation (p=0.004). Maximum pre-partum pregnane levels did
not affect time to ovulation (p=0.34). In experiment 2 equine anterior pituitary
glands were harvested and the cells plated to begin a primary cell culture. After
attachment, the cells were divided into treatment groups: P4, 5��, ����, 20��, 3�� or a
control and each group subjected to a 1.0nM Gonadotropin Hormone Releasing
Hormone (GnRH) challenge. Subsequently cells and medium were collected and
analyzed for LH and FSH content using radioimmunoassay (RIA). The cells did
exhibit a response to GnRH (p=0.015 between positive and negative controls) and
there was a treatment effect for FSH (p=0.0058); only 3�� resulted in significantly
more FSH release than the positive control (p=0.043) after stimulation with GnRH.
There was no treatment effect on LH (p=0.56). Experiment 3 analyzed the
response of equine uterine myometrial tissue to pregnane treatment. Myometrial
tissue was harvested and placed in a 37��C Krebs buffered saline bath, connected to
a physiograph and repetitive spontaneous smooth muscle contraction was induced with oxytocin. Tissue was then treated with P4, 5��, ����, 20��, 3�� or a control
(ethanol). The amplitude and frequency of the spontaneous contractions were
measured and compared to the control. There were no differences between post
treatment responses of the control and pregnane treated samples in either frequency
(p=0.78) or amplitude (p=0.63) of myometrial contractions. From these data we
conclude that in vivo there is a significant and differential physiologic relationship
between pre-partum pregnanes and gonadotropins. Due to lack of response it is
unlikely that pregnanes inhibit pituitary secretion, and thus may exert their effects
elsewhere, such as at the hypothalamus. The involvement of pregnanes in
modulating myometrial contractions remains unclear. It is likely that P4 does play
a role in decreasing myometrial responses to OT, however, that result was
inconsistent in this study. / Graduation date: 2003

Identiferoai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/31549
Date29 August 2002
CreatorsStrooband, Jenny A.
ContributorsHoltan, Donald W.
Source SetsOregon State University
Languageen_US
Detected LanguageEnglish
TypeThesis/Dissertation

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