Secretion of cumulus expansion-enabling factor (CEEF) in porcine follicles

The objective of this study was to find out whether porcine cumulus and mural granulosa cells can secrete cumulus expansion‐enabling factor (CEEF). Culture drops of M‐199 medium were conditioned with denuded porcine oocytes (1 oocyte/μl), cumulus cells from oocytectomized complexes (1 OOX/μl), piece...

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Published in:Molecular reproduction and development 1998-02, Vol.49 (2), p.141-149
Main Authors: Prochazka, R, Nagyova, E, Brem, G, Schellander, K, Motlik, J
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Communication - physiology
Cell Division - physiology
Cell Separation
Cells, Cultured
CELLULE DE LA GRANULOSA
CELULAS DE LA GRANULOSA
CERDO
CONTROL HORMONAL
CONTROLE HORMONAL
Culture Media, Conditioned
cumulus
CUMULUS OOPHORUS
expansion
EXPERIMENTACION IN VITRO
EXPERIMENTATION IN VITRO
Fallopian Tubes - secretion
Female
FOLICULOS OVARICOS
FOLLICULE OVARIEN
FSH
Fundamental and applied biological sciences. Psychology
GRANULOSA CELLS
Granulosa Cells - metabolism
Granulosa Cells - physiology
Growth Substances - secretion
HFS
HORMONAL CONTROL
Hormone metabolism and regulation
Humans
IN VITRO EXPERIMENTATION
Mammalian female genital system
Mice
oocyte
OOCYTES
Oocytes - cytology
Oocytes - physiology
Oocytes - secretion
OVA
Ovarian Follicle - cytology
Ovarian Follicle - secretion
OVARIAN FOLLICLES
OVULE
OVULO
pig
PORCIN
SECRECION
SECRETION
SWINE
Vertebrates: reproduction
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Summary:The objective of this study was to find out whether porcine cumulus and mural granulosa cells can secrete cumulus expansion‐enabling factor (CEEF). Culture drops of M‐199 medium were conditioned with denuded porcine oocytes (1 oocyte/μl), cumulus cells from oocytectomized complexes (1 OOX/μl), pieces of mural granulosa isolated from preantral to preovulatory follicles (1000 cells/μl), or oviductal cells (1000 cells/μl) for 24 hr. The production of CEEF was assessed by the addition of mouse OOX and follicle‐stimulating hormone (FSH) (1 μg/ml) to microdrops of the conditioned medium. After 16–18 hr, expansion of the mouse OOX was scored on a scale of 0 to 4 by morphologic criteria. Mouse OOX did not expand in nonconditioned FSH‐supplemented medium. Immature porcine oocytes produced +3 to +4 expansion of the mouse OOX. Granulosa cells isolated from preantral and early antral follicles and cumulus cells isolated from all stages of follicle development constitutively secreted CEEF under in vitro conditions. Mural granulosa cells of small, medium, and preovulatory (PMSG) follicles also secreted CEEF in vitro; however, FSH or leutenizing hormone (LH) stimulation was essential for this secretion. Hormonally induced secretion of CEEF was accompanied by expansion of the mural granulosa itself. Granulosa cells isolated from follicles of gilts 20 hr after PMSG and human chorionic gonadotropin (hCG) administration did not produce CEEF and did not expand in response to FSH and LH in vitro. CEEF activity also was found in the follicular fluid of small antral follicles, was reduced in medium follicles, and was not detectable in PMSG‐stimulated follicles. However, CEEF activity was reestablished in the follicular fluid of preovulatory follicles by hCG injection, conceivably due to increased production of CEEF by cumulus cells. We conclude that (1) porcine cumulus and mural granulosa cells are capable of CEEF production in vitro and (2) autocrine secretion of CEEF by cumulus cells is involved in regulation of porcine cumulus expansion both in vitro and in vivo. Mol. Reprod. Dev. 49:141–149, 1998. © 1998 Wiley‐Liss, Inc.
ISSN:1040-452X
1098-2795
DOI:10.1002/(SICI)1098-2795(199802)49:2<141::AID-MRD5>3.0.CO;2-P