mRNA expression profile of the TNF-α system in LH-induced bovine preovulatory follicles and effects of TNF-α on gene expression, ultrastructure and expansion of cumulus-oocyte complexes cultured in vitro.
This study evaluated (1) the effects of in vivo GnRH treatment on mRNA expression of TNF-α system (TNF-α, TNFR1 and TNFR2) in granulosa cells of bovine preovulatory follicles, (2) the in vitro influence of gonadotropins on mRNA expression of TNF-α system in cultured cumulus cells, (3) the protein expression of the TNF-α system in late antral follicles and, (4) the influence of TNF-α on cumulus cells expansion, ultrastructure and on expression of HAS2, CASP3 and CASP6 in follicular cells cultured for 24 h. An increased expression of TNF-α and TNFR1 was observed after 3, 6 and 12 h of GnRH treatment when compared to 0 and 24h. Higher TNFR2 mRNA levels were observed 3, 6 and 12 h after GnRH, when compared to 0 and 24 h. Proteins of TNF-α system were also expressed in late antral follicles. In vitro, TNF-α did not affect cumulus cells expansion, but reduced the HAS2, CASP3 and CASP6 mRNA levels in cumulus cells after 12 h. After 24 h of culture, TNF-α increased the mRNA levels for CASP6 in mural granulosa cells, while the TNF-α, TNFR1 and TNFR2 mRNA levels were increased in cumulus-oocyte complexes (COCs) cultured for 12 h with gonadotropins, but not after 24 h. Ultrastructural analysis confirmed the integrity of COCs cultured in presence of TNF-α. In conclusion, TNF-α system members are present in bovine antral follicles and expression of TNF-α is influenced by gonadotropins in vivo and in vitro. In vitro, TNF-α maintained cumulus cells ultrastructure during COC culture.
Biotechnology Nucleus of Sobral - NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
A W B
AWB
Silva
Biotechnology Nucleus of Sobral - NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
F T G
FTG
Bezerra
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.
W G
WG
Glanzner
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.
J T
JT
Dos Santos
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.
A M P
AMP
Dau
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.
M T
MT
Rovani
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.
G F
GF
Ilha
Biotechnology Nucleus of Sobral - NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
J J N
JJN
Costa
Biotechnology Nucleus of Sobral - NUBIS, Federal University of Ceara, Sobral, CE, Brazil.
E V
EV
Cunha
Laboratory of Ultrastructure, CPqAM/FIOCRUZ, Federal University of Pernambuco, Recife, PE, Brazil.
M A M
MAM
Donato
Laboratory of Ultrastructure, CPqAM/FIOCRUZ, Federal University of Pernambuco, Recife, PE, Brazil.
C A
CA
Peixoto
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.
P B D
PBD
Gonçalves
Department of Animal Science, McGill University, Ste-Anne-De-Bellevue, QC, Canada.
V
V
Bordignon
Biotechnology Nucleus of Sobral - NUBIS, Federal University of Ceara, Sobral, CE, Brazil. Electronic address: jrvsilva@ufc.br.
J R V
JRV
Silva
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D054885:Cumulus Cells / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D015870:Gene Expression; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D007986:Luteinizing Hormone / Q000378:metabolism* / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics / Q000378:metabolism*; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
D012333:RNA, Messenger; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha; D007987:Gonadotropin-Releasing Hormone; D007986:Luteinizing Hormone
Cattle; Cell culture; Cytokine; Ovary; Ovulation; Protein