Expression of membrane-type matrix metalloproteinases (MT) and tissue
inhibitors of matrix metalloproteinases (TIMP) was evaluated in sheep embryos and
uterus during the pre- and peri-implantation periods. Embryos and uterine samples were
surgically collected from ewes on days 9, 11, 13, and 15 of pregnancy (n=3 ewes/day)
and of the estrous cycle (n=2 ewes/day). Total RNA was extracted and RT-PCR were
performed using primers specifically designed from published human, mouse, and bovine
complete cDNA sequences for MT-1, -2, -3, and -5, and TIMP-1, -2 and -3. Multiplex
PCR were performed on uterine samples for each gene at optimal cycles and
temperatures with 18S rRNA as the internal standard. For embryos, PCR were conducted
for 40 cycles at optimal temperatures. MT-1, -2, -3, and -5 were observed in pregnant
and nonpregnant uterus during all days of collection. No difference (P>0.10) was
observed in MT-1 or -2 expression due to day of collection. However, pregnant uterus
expressed more (P=0.096) MT-1 than nonpregnant uterus, whereas expression of MT-2
was greater (P<0.05) in nonpregnant compared to pregnant uterus. No differences (P>0.10) in MT-3 expression were observed due to pregnancy status, however Day 9 and 11
expressed more MT-3 than Day 15. Uterine MT-5 expression was not different (P>0.10) between pregnant and nonpregnant females, however Day 15 uterus expressed less
(P<0.05) MT-5 then Day 11 and 13 uteri. TIMP-1 expression was greater (P<0.05) in
pregnant compared to nonpregnant uterus, but did not differ (P>0.10) by day of
collection. TIMP-2 did not differ (P>0.10) by pregnancy status or day of collection but
the interaction was significant (P<0.05). TIMP-2 expression was greatest in Day 9
pregnant uterus and least in Day 9 nonpregnant uterus. No difference (P>0.10) was
observed in expression of TIMP-3 due to day of collection or pregnancy status. Embryos
expressed MT-3 and -5 during Days 9-15 of development, however, MT-1 and -2 were
not detected. The presence of MT and TIMP in the endometrium suggests these proteins
may play important roles in regulating extracellular matrix degradation and activating
other matrix metalloproteinases for endometrial remodeling and preparation for
implantation. Embryonic MT may participate in the processes of embryonic expansion,
elongation and attachment. / Graduation date: 2002
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/29977 |
| Date | 05 October 2001 |
| Creators | Paul, Katy Beth |
| Contributors | Menino, Alfred R. Jr |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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