Membrane-type matrix metalloproteinase and inhibitor expression in sheep embryos and uterus Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/st74cv055

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  • 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.
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