Role of fibroblast growth factor 18 in regulating the cascade of pre-ovulatory events in cattle

Reis, Dalila do Nascimento; Missio, Daniele; Fiorenza, Mariani Farias; Antoniazzi, Alfredo Quites; Price, Christopher; Gonçalves, Paulo Bayard Dias; Portela, Valério Marques

doi:10.1590/0103-8478cr20220215

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ANIMAL REPRODUCTION • Cienc. Rural 54 (5) • May 2024 • https://doi.org/10.1590/0103-8478cr20220215 copy

Role of fibroblast growth factor 18 in regulating the cascade of pre-ovulatory events in cattle

Função do fator de crescimento fibroblástico 18 na regulação da cascata de eventos pré-ovulatórios em bovinos

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT:

Suboptimal LH surges can lead to delayed or absent ovulation during reproductive management programs in cattle. Several signaling pathways may enhance the LH signal, potentially including growth factors. In the present study, we determined whether Fibroblast growth factor 18 (FGF18), which mimics some actions of the LH surge on granulosa cells, may potentiate the action of a suboptimal dose of LH using an established bovine granulosa cell (GC) culture system. Addition of recombinant FGF18 increased the concentration of progesterone (P4) in the culture medium. A combination of LH and FGF18 in the culture medium increased the abundance of mRNA encoding EREG, EGR1 and EGR3, proteins that are early modulators of the LH-induced pre-ovulatory cascade. This study demonstrated that FGF18 may be a co-factor in the regulation of the pre-ovulatory cascade in bovine ovarian granulosa cells.

Key words:
FGF18; ovulation; granulosa cells

Authorship

Dalila do Nascimento Reis

conceptualization

Writing-original draft

Writing-review and editing

Programa de Pós-graduação em Medicina Veterinária, Centro de Ciências Rurais (CCR), Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brasil.Universidade Federal de Santa Maria (UFSM)BrazilSanta Maria, RS, BrazilPrograma de Pós-graduação em Medicina Veterinária, Centro de Ciências Rurais (CCR), Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brasil.

https://orcid.org/0000-0001-7680-5153

Daniele Missio

conceptualization

Writing-original draft

Writing-review and editing

https://orcid.org/0000-0003-3524-1894

Mariani Farias Fiorenza

conceptualization

Writing-original draft

Writing-review and editing

https://orcid.org/0000-0001-8202-0678

Alfredo Quites Antoniazzi

conceptualization

Writing-original draft

Writing-review and editing

Laboratório de Biotecnologia e Reprodução Animal (BioRep), Departamento de Fisiologia e Farmacologia, Centro de Ciências da Saúde, Universidade Federal de Santa Maria (UFSM), 97105-900, Santa Maria, RS, Brasil..Universidade Federal de Santa Maria (UFSM)BrazilSanta Maria, RS, BrazilLaboratório de Biotecnologia e Reprodução Animal (BioRep), Departamento de Fisiologia e Farmacologia, Centro de Ciências da Saúde, Universidade Federal de Santa Maria (UFSM), 97105-900, Santa Maria, RS, Brasil..

https://orcid.org/0000-0003-4866-5944

Christopher Price

conceptualization

Writing-original draft

Writing-review and editing

Centre de recherche en reproduction et fertilité (CRRF), Faculté de médecine Vétérinaire (FMV), Université de Montréal (UdeM), Montreal, Canada.Université de Montréal (UdeM)CanadaMontreal, CanadaCentre de recherche en reproduction et fertilité (CRRF), Faculté de médecine Vétérinaire (FMV), Université de Montréal (UdeM), Montreal, Canada.

https://orcid.org/0000-0002-6727-8280

Paulo Bayard Dias Gonçalves

conceptualization

Writing-original draft

Writing-review and editing

https://orcid.org/0000-0002-0671-1466

Valério Marques Portela ^*E-mail: valerio.portela@ufsm.br. *Corresponding author

conceptualization

Writing-original draft

Writing-review and editing

https://orcid.org/0000-0003-2109-1346

E-mail: valerio.portela@ufsm.br. ^*Corresponding author

Editor: Rudi Weiblen (0000-0002-1737-9817)

DECLARATION OF CONFLICT OF INTEREST

The authors declare no conflicts of interest.

AUTHORS’ CONTRIBUTIONS

All authors contributed equally to the conception and writing of the manuscript. All authors critically revised the manuscript and approved the final version.

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (3)
Tables (1)

Thumbnail

Figure 1
Effects of FGF18 and LH on the secretion of progesterone (P4) from bovine granulose cells. Cells were cultured for 18 h in serum-free medium without supplementation and collected at 6, 12, and 24 h after adding FGF18 and LH (10 ng/ml) to treatment groups. Data are presented as means  SEM of three independent replicates. The asterisk denote statistical differences between the groups by time (^*P < 0.05).

Thumbnail

Figure 2
Effects of FGF18 and LH on the expression of genes critical for ovulation. Cells from large follicles (≥10 mm) were cultured for 18 h in serum-free medium without supplementation and collected at 6, 12, and 24 h after adding FGF18 and LH (10 ng/ml) to treatment groups. mRNA abundance was measured using real-time PCR. Data are presented as means ± SEM of three independent replicates. Different letters show statistically significant differences between all treatment means by time within this experiment (P < 0.05).

Thumbnail

Figure 3
Effects of FGF18 and LH on the expression of the Hippo pathway target genes during the pre-ovulatory cascade. Cells from large follicles (≥10 mm) were cultured for 18 h in serum-free medium without supplementation and collected at 6, 12, and 24 h after adding FGF18 and LH (10 ng/ml) to treatment groups. mRNA abundance was measured using real-time PCR. Data are presented as means ? SEM of three independent replicates. Different letters show statistically significant differences between all treatment means by time within this experiment (P < 0.05).

Thumbnail

Table 1
Gene and primers sequence used.

Figure 1 Effects of FGF18 and LH on the secretion of progesterone (P4) from bovine granulose cells. Cells were cultured for 18 h in serum-free medium without supplementation and collected at 6, 12, and 24 h after adding FGF18 and LH (10 ng/ml) to treatment groups. Data are presented as means  SEM of three independent replicates. The asterisk denote statistical differences between the groups by time (^*P < 0.05).

Figure 2 Effects of FGF18 and LH on the expression of genes critical for ovulation. Cells from large follicles (≥10 mm) were cultured for 18 h in serum-free medium without supplementation and collected at 6, 12, and 24 h after adding FGF18 and LH (10 ng/ml) to treatment groups. mRNA abundance was measured using real-time PCR. Data are presented as means ± SEM of three independent replicates. Different letters show statistically significant differences between all treatment means by time within this experiment (P < 0.05).

Figure 3 Effects of FGF18 and LH on the expression of the Hippo pathway target genes during the pre-ovulatory cascade. Cells from large follicles (≥10 mm) were cultured for 18 h in serum-free medium without supplementation and collected at 6, 12, and 24 h after adding FGF18 and LH (10 ng/ml) to treatment groups. mRNA abundance was measured using real-time PCR. Data are presented as means ? SEM of three independent replicates. Different letters show statistically significant differences between all treatment means by time within this experiment (P < 0.05).

Table 1 Gene and primers sequence used.

Gene symbol	-----------------------Forward primer----------------------	-----------------------Reverse primer----------------------
H2AFZ	GAGGAGCTGAACAAGCTGTTG	TTGTGGTGGCTCTCAGTCTTC
EGR1	TCCCCTGTTCACAATGGTTT	TGGGAGAAAAGGTGGTTGTC
EGR3	GGAGCAAATGAAATGTTGGTG	AGGAAAACCTATGGGGAATG
AREG	CTTTCGTCTCTGCCATGACCTT	CGTTCTTCAGCGACACCTTCA
EREG	ACTGCACAGCATTAGTTCAAACTGA	TGTCCATGCAAACAGTAGCCATT
CTGF	AGCTGAGCGAGTTGTGTACC	TCCGAAAATGTAGGGGGCAC
CYR61	GGCTCCCCGTTTTGGAATG	TCATTGGTAACGCGTGTGGA
BIRC5	CTGAGAACGAGCCCGACTTG	ATGTTCTTCTATAGGGTCGTCATCT
ANKRD1	ATCAGTGCGCGGGATAAGTT	GGGAGTATCTCCTTCCCGGT
YAP1	TCCTTTGAGATCCCTGACGATG	TGACGTTCATCTGGGAGAGC
COX-2	CCTGTGTTCCACCAGGAGAT	CCCTGGCTAGTGCTTCAGAC

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Universidade Federal de Santa Maria Universidade Federal de Santa Maria, Centro de Ciências Rurais , 97105-900 Santa Maria RS Brazil , Tel.: +55 55 3220-8698 , Fax: +55 55 3220-8695 - Santa Maria - RS - Brazil
E-mail: cienciarural@mail.ufsm.br

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[1] E-mail: valerio.portela@ufsm.br. ^*Corresponding author