Bovine Follicular Fluid and Serum Share a Unique Isoform of Matrix Metalloproteinase-2 That Is Degraded by the Oviductal Fluid
Whereas the mammalian fertilization environment consists of possible products of the mutual interaction between oviductal and follicular fluids in addition to both fluid components, little is known regarding the interaction. In the present study, we have demonstrated that a mutual interaction occurs...
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Published in | Biology of reproduction Vol. 65; no. 6; pp. 1726 - 1731 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Madison, WI
Society for the Study of Reproduction
01.12.2001
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Subjects | |
Online Access | Get full text |
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Summary: | Whereas the mammalian fertilization environment consists of possible products of the mutual interaction between oviductal
and follicular fluids in addition to both fluid components, little is known regarding the interaction. In the present study,
we have demonstrated that a mutual interaction occurs, resulting in the biochemical changes of follicular fluid components.
Gelatin zymographic analyses of bovine follicular fluid (bFF) showed consistently a distinct, gelatinolytic activity having
a molecular weight of 110 kDa (GA110) in addition to other gelatinases, whereas bovine oviductal fluid (bOF) showed a lack
of GA110. Surprisingly, when bFF was mixed with bOF before zymography, the GA110 of bFF mostly disappeared at a 1:1 (v/v)
mixture, completely disappeared at a 1:10 mixture, as fast as within 30 min after mixing. Other bFF gelatinase activities
were not affected by bOF at 1:1 or 10:1 mixtures. Addition of EDTA or phenanthroline, but not of phenylmethylsulfonyl fluoride
or trypsin inhibitor, to the mixture greatly increased the gelatinolytic activity of bFF GA110. The increased activity of
bFF GA110 by EDTA was again abolished by subsequent bOF treatment. Addition of aminophenylmercuric acetate to the EDTA-treated
bFF also abolished GA110; however, this was accompanied by the disappearance of other gelatinases, except the 62-kDa gelatinase,
the activity of which increased as the treatment continued up to 24 h. Addition of EDTA or phenanthroline to the gelatin gel
incubation buffer after electrophoresis abolished almost all gelatinases of bFF, except those of 88â84 kDa, demonstrating
that they were indeed gelatinases or isoforms. Bovine serum and fetal bovine serum also showed the presence of GA110, the
activity of which was increased by EDTA. However, ovarian granulosa cell homogenate did not exhibit GA110. Immunoblot experiments
using antibodies against matrix metalloproteinase (MMP)-2 and MMP-9 demonstrated that bFF GA110 was an isoform of MMP-2, and
that the 62-kDa form was an active form of MMP-2. Disappearance of immunoreactive GA110 of bFF and serum by bOF was also observed.
Based on these observations, we conclude that bFF and bovine serum share a unique isoform of MMP-2, and that bOF can specifically
degrade the isoform, suggesting that a mutual interaction between bFF and bOF could occur at the time of ovulation. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0006-3363 1529-7268 |
DOI: | 10.1095/biolreprod65.6.1726 |