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Evidence that Melatonin Increases Inhibin Beta-A and Follistatin Gene Expression in Ovaries of Pinealectomized Rats

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Abstract

Melatonin plays an important role in the regulation of ovarian function including oocyte maturation in different mammalian species. Many studies indicate that melatonin has an impact on the ovarian function of a variety of ovarian cells. However, the information on the exact mechanism and involved hormones is low. To evaluate inhibin beta-A (INHBA) and follistatin (FST) expression in the ovaries of pinealectomized rats treated with melatonin, thirty adult female Wistar rats were randomized into three groups of ten animals each: group 1 (GSh), sham-operated controls receiving vehicle; group 2 (GPx), pinealectomized animals receiving vehicle; and group 3 (GPxMe), pinealectomized animals receiving replacement melatonin (1.0 mg/kg body weight. It was assumed that each animal drank 6.5 ± 1.2 ml per night and weighs approximately 300 g.) for 60 consecutive days. The ovaries were collected for mRNA abundance and protein of INHBA and FST by qRT-PCR and immunohistochemical analyses, respectively. Treatment with melatonin resulted in the upregulation of INHBA and FST genes in the ovarian tissue of the melatonin-treated animals (GPxMe), when compared with GPx. These findings were then confirmed by analyzing the expression of protein by immunohistochemical analyses, which revealed higher immunoreactivity of INHBA and FST in GPxMe animals in the follicular cells compared with GSh and GPx rats. Melatonin increases the expression of INHBA and FST in the ovaries of pinealectomized female rats.

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Acknowledgments

Thank you for reviewing the manuscript Prof. Dr. KMJ Menon and Researcher Dr. Bindu Menon of the Department of Obstetrics and Gynecology of the University of Michigan Medical School, Ann Arbor, MI, USA. Also, we thank Prof. Reiter Russel (Department of Cellular & Structural Biology, University of Texas, Health Science Center, San Antonio) for reviewing the manuscript.

Availability of Data and Material

The datasets supporting the conclusions of this article are included within the article.

Funding

This research was supported by São Paulo Research Foundation (FAPESP) (Grant Number 2011/51581-8 and 2012/50489; FAPESP Process Number 2009/54019-9), Pio XI St., 1500, Alto da Lapa, São Paulo/SP, Brazil, Postal Code 05468-901.

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Authors and Affiliations

  1. Laboratório de Investigação Médica (LIM 58) - Disciplina de Ginecologia, Departamento de Obstetrícia e Ginecologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil

    Carla C. Maganhin, Maria Cândida P. Baracat, Kátia C. Carvalho, Isadora Braga Seganfredo, Camilla Maganhin Luquetti, Ricardo dos Santos Simões, Edmund C. Baracat & José M. Soares-Jr

  2. Departamento de Morfologia e Genética, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil

    Adriana Aparecida Ferraz Carbonel & Manuel de Jesus Simões

  3. Departamento de Fisiologia e Biofísica, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil

    José Cipolla-Neto

  4. Departamento de Ginecologia, Universidade Federal de São Paulo, São Paulo, Brazil

    Manoel João Batista Castello Girão

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  1. Carla C. Maganhin
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  2. Maria Cândida P. Baracat
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  12. José M. Soares-Jr
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Contributions

CCM conceived of and designed the study, performed experiments, interpreted data, and wrote the manuscript. ECB and CCM conceived of and designed the study, corrected the manuscript, made substantial contributions to the study conception, and critically revised the manuscript for important intellectual content. CCM and MJS performed experiments and wrote specific sections of the manuscript. JCN and MCB performed experiments and interpreted morphological data. MCG, AAFC, and ECB interpreted data and corrected the manuscript. ECB and JMS designed the study and participated in the revision of the manuscript. All the authors read and approved the final manuscript.

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Correspondence to José M. Soares-Jr.

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This project was approved by the Institutional Research Committee at Federal University of São Paulo (0233/06), in accordance with the National Institutes of Health (NIH) Guidelines Regarding Animal Experimentation and with the ethical principles recommended by the Brazilian Committee on Animal Experimentation (COBEA, Brazil).

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Where the work was performed: Laboratório de Investigação Médica (LIM-58) da Disciplina de Ginecologia do Departamento de Obstetrícia e Ginecologia da Faculdade de Medicina da Universidade de São Paulo/Laboratório de Histologia e Biocelular do Departamento de Morfologia e Genética da Universidade Federal Sao Paulo.

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Maganhin, C.C., Baracat, M.C.P., Carvalho, K.C. et al. Evidence that Melatonin Increases Inhibin Beta-A and Follistatin Gene Expression in Ovaries of Pinealectomized Rats. Reprod. Sci. 27, 1455–1464 (2020). https://doi.org/10.1007/s43032-020-00162-1

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