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The effects of the obesogen tributyltin on the metabolism of Sertoli cells cultured ex vivo

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Abstract

Human exposure to environmental contaminants is widespread. Some of these contaminants have the ability to interfere with adipogenesis, being thus considered as obesogens. Recently, obesogens have been singled out as a cause of male infertility. Sertoli cells (SCs) are essential for male fertility and their metabolic performance, especially glucose metabolism, is under a tight endocrine control, being essential for the success of spermatogenesis. Herein, we studied the impact of the model obesogen tributyltin in the metabolic profile of SCs. For that, ex vivo-cultured rat SCs were exposed to increasing doses of tributyltin. SCs proliferation was evaluated by the sulforhodamine B assay and the maturation state of the cells was assessed by the expression of specific markers (inhibin B and the androgen receptor) by quantitative polymerase chain reaction. The metabolic profile of SCs was established by studying metabolites consumption/production by nuclear magnetic resonance spectroscopy and by analyzing the expression of key transporters and enzymes involved in glycolysis by Western blot. The proliferation of SCs was only affected in the cells exposed to the highest dose (1000 nM) of tributyltin. Notably, SCs exposed to 10 nM tributyltin decreased the consumption of glucose and pyruvate, as well as the production of lactate. The decreased lactate production hampers the development of germ cells. Intriguingly, the lowest levels of tributyltin were more prone to modulate the expression of key players of the glycolytic pathway. This is the first study showing that tributyltin reprograms glucose metabolism of SCs under ex vivo conditions, suggesting new targets and mechanisms through which obesogens modulate the metabolism of SCs and thus male (in)fertility.

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Abbreviations

1H-NMR:

Proton nuclear magnetic resonance

AR:

Androgen receptor

β2MG:

β2-Microglobulin

CAMs:

Cell adhesion molecules

cDNA:

Complementary deoxyribonucleic acid

DMEM:F12:

Dulbecco’s modified eagle medium Ham’s nutrient mixture F12

EtOH:

Ethanol

GLUT1:

Glucose transporter 1

GLUT2:

Glucose transporter 2

GLUT3:

Glucose transporter 3

GLUTs:

Glucose transporters

HBSS:

Hank’s balanced salts solution

ITS:

Insulin–transferrin–sodium selenite

LDH:

Lactate dehydrogenase

MCT4:

Monocarboxylate transporter 4

PBS:

Phosphate-buffered Saline

PFK1:

Phosphofructokinase 1

PPARy:

Peroxisome proliferator-activated receptor gamma

RXR:

Retinoid X receptor

SCs:

Sertoli cells

SRB:

Colorimetric sulforhodamine B

T:

Testosterone

TBT:

Tributyltin

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Acknowledgements

This work is supported by FEDER funds through the POCI—COMPETE 2020—Operational Programme Competitiveness and Internationalisation in Axis I—Strengthening research, technological development and innovation (Project No. 007491) and National Funds by FCT—Foundation for Science and Technology (Project UID/Multi/00709); CICECO (Ref. FCT UID/CTM/50011/2013); UMIB (PEst-OE/SAU/UI0215/2014). NMR data was collected at the UC-NMR facility which is supported in part by FEDER—European Regional Development Fund through the COMPETE Programme (Operational Programme for Competitiveness) and by National Funds through FCT—Fundação para a Ciência e a Tecnologia (Portuguese Foundation for Science and Technology) through Grants REEQ/481/QUI/2006, RECI/QEQ-QFI/0168/2012, CENTRO-07-CT62-FEDER-002012, and Rede Nacional de Ressonância Magnética Nuclear (RNRMN). P.F. Oliveira (PTDC/BBB-BQB/1368/2014 and IFCT2015); M.G. Alves (IFCT2015 and PTDC/BIM-MET/4712/2014)) and L. Rato was funded by Santander-Totta/University of Beira Interior (BIPD/ICI-CICS-BST-UBI).

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Authors and Affiliations

  1. Health Sciences Research Centre (CICS–UBI), University of Beira Interior, Rua Marquês d’Ávila e Bolama, Av. Infante D. Henrique, 6200-001, Covilhã, Portugal

    Ana M. Cardoso, José E. Cavaco & Luís Rato

  2. Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal

    Marco G. Alves, Ivana Jarak & Pedro F. Oliveira

  3. CICECO - Aveiro Institute of Materials, Chemistry Department, University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal

    Ana C. Sousa

  4. Department of Life Sciences, Faculty of Sciences and Technology and Centre for Functional Ecology (CFE), University of Coimbra, Coimbra, Portugal

    Ivana Jarak & Rui A. Carvalho

  5. i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal

    Pedro F. Oliveira

  6. Department of Genetics, Faculty of Medicine, University of Porto, Porto, Portugal

    Pedro F. Oliveira

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  1. Ana M. Cardoso
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  2. Marco G. Alves
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  3. Ana C. Sousa
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  4. Ivana Jarak
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  6. Pedro F. Oliveira
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  7. José E. Cavaco
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  8. Luís Rato
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Correspondence to Luís Rato.

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Cardoso, A.M., Alves, M.G., Sousa, A.C. et al. The effects of the obesogen tributyltin on the metabolism of Sertoli cells cultured ex vivo. Arch Toxicol 92, 601–610 (2018). https://doi.org/10.1007/s00204-017-2091-x

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