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The Hippo pathway effectors YAP and TAZ interact with EGF-like signaling to regulate expansion-related events in bovine cumulus cells in vitro

  • Reproductive Physiology and Disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract  

Purpose

To determine if the inhibition of the interaction between the Hippo effector YAP or its transcriptional co-activator TAZ with the TEAD family of transcription factors is critical for the cumulus expansion–related events induced by the EGF network in cumulus-oocyte complexes (COCs).

Methods

We performed a series of experiments using immature bovine COCs subjected to an IVM protocol for up 24 h in which cumulus expansion was stimulated with EGF recombinant protein or FSH.

Results

The main results indicated that EGFR activity stimulation in bovine cumulus cells (CC) increases mRNA levels encoding the classic YAP/TAZ-TEAD target gene CTGF. To determine if important genes for cumulus expansion are transcriptional targets of YAP/TAZ-TEAD interaction in CC, COCs were then subjected to IVM in the presence of FSH with or without distinct concentrations of Verteporfin (VP; a small molecule inhibitor that interferes with YAP/TAZ binding to TEADs). COCs were then collected at 6, 12, 18, and 24 h for total RNA extraction and RT-qPCR analyses. This experiment indicated that VP inhibits in a time- and concentration-dependent manner distinct cumulus expansion and oocyte maturation–related genes, by regulating EGFR and CTGF expression in CC.

Conclusions

Taken together, the results presented herein represent considerable insight into the functional relevance of a completely novel signaling pathway underlying cumulus expansion and oocyte maturation in monovulatory species. YAP/TAZ or CTGF may represent potential targets to improve the efficiency of IVM systems, not only for monovulatory species of agricultural importance as the cow, but for human embryo production.

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Data availability

The data sets used and/or analyzed during the current study are available from the corresponding author on reasonable request. All data generated or analyzed during this study are included in this published article.

Code availability

Not applicable.

Abbreviations

ADAM17:

A disintegrin and metalloproteinase 17

AREG:

Amphiregulin

CC:

Cumulus cells

COCs:

Cumulus-oocyte complexes

CTGF:

Connective tissue growth factor

DMSO:

Dimethyl sulfoxide

EGF:

Epidermal growth factor

EGFR:

Epidermal growth factor receptor

EREG:

Epiregulin

FSH:

Follicle-stimulating hormone

GC:

Granulosa cells

H2AFZ:

Histone H2A.Z

HAS2:

Hyaluronan synthase 2

IHC:

Immunohistochemistry

IVM:

In vitro maturation

LH:

Luteinizing hormone

PLAT:

Plasminogen activator tissue-type A

PTGS2:

Prostaglandin endoperoxide synthase 2

PTX3:

Pentraxin-related protein 3

RPS18:

Ribosomal protein S18

TAZ:

Transcriptional co-activator with PDZ-binding motif

TEAD:

Transcriptional enhanced associate domain

VP:

Verteporfin

YAP:

Yes-associated protein 1

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Acknowledgements

The authors are thankful to Mrs. Camila Azzolin de Souza and Mrs. Camila Cupper for their support in this study and to Frigorífico Silva for ovaries donation.

Funding

This work was supported by Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant RGPIN-2018–06470 (to Dr. Zamberlam) and by fellowships from the National Council for Scientific and Technological Development (CNPq; Brazil), Coordination for the Improvement of Higher Education Personnel (CAPES; Brazil) and by the grants from Rio Grande do Sul State Research Support Foundation (FAPERGS – Edital 06/2019; 19/2551–0002275-1) & CNPq, Edital 16/2551–0000494-3; Brazil (to Dr. Gonçalves).

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Authors and Affiliations

  1. Centre de Recherche en Reproduction et Fertilité (CRRF), Faculté de Médecine Vétérinaire (FMV), Université de Montréal (UdeM), 3200 rue Sicotte, St-Hyacinthe, QC, J2S 7C6, Canada

    Júlia Koch, Esdras Corrêa Dos Santos & Gustavo Zamberlam

  2. Laboratory of Biotechnology and Animal Reproduction, BioRep, Veterinary Hospital, Federal University of Santa Maria (UFSM), Santa Maria, RS, 97105-900, Brazil

    Júlia Koch, Valério Marques Portela, Daniele Missio, Leonardo Guedes de Andrade, Zigomar da Silva, Alfredo Quites Antoniazzi & Paulo Bayard Dias Gonçalves

  3. Federal University Pelotas (UFPel), Pelotas, RS, 96010-900, Brazil

    Bernardo Garziera Gasperin

  4. Federal University of Pampa (Unipampa), Uruguaiana, RS, 97501-970, Brazil

    Paulo Bayard Dias Gonçalves

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  1. Júlia Koch
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  2. Valério Marques Portela
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  4. Daniele Missio
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  10. Gustavo Zamberlam
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Contributions

J.K., V.M.P., P.B.D.G., and G.Z. were involved in the study conception and design; J.K., V.M.P., E.C., D.M., L.G.A., Z.S., and A.Q.A. performed experiments and were involved in the acquisition and analyzes of data; G.Z., P.B.D.G., and B.G.G contributed with required reagents acquisition; J.K. wrote the main manuscript text and prepared the tables and figures; G.Z. and P.B.D.G. reviewed and edited the manuscript to be published.

Corresponding author

Correspondence to Gustavo Zamberlam.

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All ovaries were obtained from a local slaughterhouse and in vitro procedures were approved by the Ethics and Animal Welfare Committee of the Federal University of Santa Maria, Brazil/not applicable.

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The authors declare no competing interests.

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Koch, J., Portela, V.M., Dos Santos, E.C. et al. The Hippo pathway effectors YAP and TAZ interact with EGF-like signaling to regulate expansion-related events in bovine cumulus cells in vitro. J Assist Reprod Genet 39, 481–492 (2022). https://doi.org/10.1007/s10815-021-02384-x

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