Thyroid hormones act as mitogenic and pro survival factors in rat ovarian follicles

R Canipari, C Mangialardo, V Di Paolo, F Alfei, S Ucci, V Russi, M G Santaguida, C Virili, M Segni, S Misiti, M Centanni, C Verga Falzacappa

Research output: Contribution to journalArticle

Abstract

PURPOSE: Thyroid disorders are clinically associated with impaired fertility in women, and these abnormalities can be improved by restoring the euthyroid state. The exact mechanisms of thyroid effect on female fertility are not well known; however, it is conceivable that thyroid hormones (THs) might act on ovarian physiology via receptors in granulosa cells. This work is aimed at evaluating the effects of THs on non-tumoral granulosa cells and follicles.

METHODS: Freshly isolated rat ovarian follicles and granulosa cells were exposed to T3 or T4 (THs). Cell growth and viability were evaluated by cell counting and the MTT assay, respectively, follicle growth was evaluated by volume measurements. Apoptosis was evaluated by the TUNEL assay and active Caspase 3 staining. rGROV cells were exposed to T3, and apoptosis was induced by serum deprivation. Bcl2, Bcl-2-associated X protein (BAX), Akt and pAkt expression were evaluated by western blot.

RESULTS: T3 induced a 40% increase in follicle volume (after 7 days). This increase was presumably due to the observed decrease (33%) in the apoptotic rate of the granulosa cell population. Both T3 and T4 caused a dose-dependent increase in rat granulosa cell number and viability. In addition, THs decreased the cell apoptotic rate in a dose-dependent manner. In both conditions, T3 appeared to be more efficient. In rGROV cells, 100 nM T3 induced cell growth and, in the absence of growth factors, reduced cell apoptosis by 40%, downregulating Caspase 3 and BAX. This effect was associated with an increase in pAkt levels. The involvement of the PI3 K pathway was confirmed by the ability of the PI3 K specific inhibitor (LY-294,002) to abolish T3 pro-survival action.

CONCLUSIONS: THs influence cell survival of ovarian granulosa cells. This effect likely contributes to the TH-induced follicle volume increase.

Original languageEnglish
JournalJournal of Endocrinological Investigation
DOIs
Publication statusE-pub ahead of print - Jun 22 2018

Fingerprint

Ovarian Follicle
Granulosa Cells
Thyroid Hormones
Survival
bcl-2-Associated X Protein
Cell Survival
Apoptosis
Caspase 3
Fertility
Thyroid Gland
Growth
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
In Situ Nick-End Labeling
Thyroxine
Intercellular Signaling Peptides and Proteins
Down-Regulation
Cell Count
Western Blotting
Staining and Labeling
Serum

Cite this

Thyroid hormones act as mitogenic and pro survival factors in rat ovarian follicles. / Canipari, R; Mangialardo, C; Di Paolo, V; Alfei, F; Ucci, S; Russi, V; Santaguida, M G; Virili, C; Segni, M; Misiti, S; Centanni, M; Verga Falzacappa, C.

In: Journal of Endocrinological Investigation, 22.06.2018.

Research output: Contribution to journalArticle

Canipari, R, Mangialardo, C, Di Paolo, V, Alfei, F, Ucci, S, Russi, V, Santaguida, MG, Virili, C, Segni, M, Misiti, S, Centanni, M & Verga Falzacappa, C 2018, 'Thyroid hormones act as mitogenic and pro survival factors in rat ovarian follicles', Journal of Endocrinological Investigation. https://doi.org/10.1007/s40618-018-0912-2
Canipari, R ; Mangialardo, C ; Di Paolo, V ; Alfei, F ; Ucci, S ; Russi, V ; Santaguida, M G ; Virili, C ; Segni, M ; Misiti, S ; Centanni, M ; Verga Falzacappa, C. / Thyroid hormones act as mitogenic and pro survival factors in rat ovarian follicles. In: Journal of Endocrinological Investigation. 2018.
@article{82e5371a370e46f187b0fe134b00fe44,
title = "Thyroid hormones act as mitogenic and pro survival factors in rat ovarian follicles",
abstract = "PURPOSE: Thyroid disorders are clinically associated with impaired fertility in women, and these abnormalities can be improved by restoring the euthyroid state. The exact mechanisms of thyroid effect on female fertility are not well known; however, it is conceivable that thyroid hormones (THs) might act on ovarian physiology via receptors in granulosa cells. This work is aimed at evaluating the effects of THs on non-tumoral granulosa cells and follicles.METHODS: Freshly isolated rat ovarian follicles and granulosa cells were exposed to T3 or T4 (THs). Cell growth and viability were evaluated by cell counting and the MTT assay, respectively, follicle growth was evaluated by volume measurements. Apoptosis was evaluated by the TUNEL assay and active Caspase 3 staining. rGROV cells were exposed to T3, and apoptosis was induced by serum deprivation. Bcl2, Bcl-2-associated X protein (BAX), Akt and pAkt expression were evaluated by western blot.RESULTS: T3 induced a 40{\%} increase in follicle volume (after 7 days). This increase was presumably due to the observed decrease (33{\%}) in the apoptotic rate of the granulosa cell population. Both T3 and T4 caused a dose-dependent increase in rat granulosa cell number and viability. In addition, THs decreased the cell apoptotic rate in a dose-dependent manner. In both conditions, T3 appeared to be more efficient. In rGROV cells, 100 nM T3 induced cell growth and, in the absence of growth factors, reduced cell apoptosis by 40{\%}, downregulating Caspase 3 and BAX. This effect was associated with an increase in pAkt levels. The involvement of the PI3 K pathway was confirmed by the ability of the PI3 K specific inhibitor (LY-294,002) to abolish T3 pro-survival action.CONCLUSIONS: THs influence cell survival of ovarian granulosa cells. This effect likely contributes to the TH-induced follicle volume increase.",
author = "R Canipari and C Mangialardo and {Di Paolo}, V and F Alfei and S Ucci and V Russi and Santaguida, {M G} and C Virili and M Segni and S Misiti and M Centanni and {Verga Falzacappa}, C",
year = "2018",
month = "6",
day = "22",
doi = "10.1007/s40618-018-0912-2",
language = "English",
journal = "Journal of Endocrinological Investigation",
issn = "0391-4097",
publisher = "Springer International Publishing",

}

TY - JOUR

T1 - Thyroid hormones act as mitogenic and pro survival factors in rat ovarian follicles

AU - Canipari, R

AU - Mangialardo, C

AU - Di Paolo, V

AU - Alfei, F

AU - Ucci, S

AU - Russi, V

AU - Santaguida, M G

AU - Virili, C

AU - Segni, M

AU - Misiti, S

AU - Centanni, M

AU - Verga Falzacappa, C

PY - 2018/6/22

Y1 - 2018/6/22

N2 - PURPOSE: Thyroid disorders are clinically associated with impaired fertility in women, and these abnormalities can be improved by restoring the euthyroid state. The exact mechanisms of thyroid effect on female fertility are not well known; however, it is conceivable that thyroid hormones (THs) might act on ovarian physiology via receptors in granulosa cells. This work is aimed at evaluating the effects of THs on non-tumoral granulosa cells and follicles.METHODS: Freshly isolated rat ovarian follicles and granulosa cells were exposed to T3 or T4 (THs). Cell growth and viability were evaluated by cell counting and the MTT assay, respectively, follicle growth was evaluated by volume measurements. Apoptosis was evaluated by the TUNEL assay and active Caspase 3 staining. rGROV cells were exposed to T3, and apoptosis was induced by serum deprivation. Bcl2, Bcl-2-associated X protein (BAX), Akt and pAkt expression were evaluated by western blot.RESULTS: T3 induced a 40% increase in follicle volume (after 7 days). This increase was presumably due to the observed decrease (33%) in the apoptotic rate of the granulosa cell population. Both T3 and T4 caused a dose-dependent increase in rat granulosa cell number and viability. In addition, THs decreased the cell apoptotic rate in a dose-dependent manner. In both conditions, T3 appeared to be more efficient. In rGROV cells, 100 nM T3 induced cell growth and, in the absence of growth factors, reduced cell apoptosis by 40%, downregulating Caspase 3 and BAX. This effect was associated with an increase in pAkt levels. The involvement of the PI3 K pathway was confirmed by the ability of the PI3 K specific inhibitor (LY-294,002) to abolish T3 pro-survival action.CONCLUSIONS: THs influence cell survival of ovarian granulosa cells. This effect likely contributes to the TH-induced follicle volume increase.

AB - PURPOSE: Thyroid disorders are clinically associated with impaired fertility in women, and these abnormalities can be improved by restoring the euthyroid state. The exact mechanisms of thyroid effect on female fertility are not well known; however, it is conceivable that thyroid hormones (THs) might act on ovarian physiology via receptors in granulosa cells. This work is aimed at evaluating the effects of THs on non-tumoral granulosa cells and follicles.METHODS: Freshly isolated rat ovarian follicles and granulosa cells were exposed to T3 or T4 (THs). Cell growth and viability were evaluated by cell counting and the MTT assay, respectively, follicle growth was evaluated by volume measurements. Apoptosis was evaluated by the TUNEL assay and active Caspase 3 staining. rGROV cells were exposed to T3, and apoptosis was induced by serum deprivation. Bcl2, Bcl-2-associated X protein (BAX), Akt and pAkt expression were evaluated by western blot.RESULTS: T3 induced a 40% increase in follicle volume (after 7 days). This increase was presumably due to the observed decrease (33%) in the apoptotic rate of the granulosa cell population. Both T3 and T4 caused a dose-dependent increase in rat granulosa cell number and viability. In addition, THs decreased the cell apoptotic rate in a dose-dependent manner. In both conditions, T3 appeared to be more efficient. In rGROV cells, 100 nM T3 induced cell growth and, in the absence of growth factors, reduced cell apoptosis by 40%, downregulating Caspase 3 and BAX. This effect was associated with an increase in pAkt levels. The involvement of the PI3 K pathway was confirmed by the ability of the PI3 K specific inhibitor (LY-294,002) to abolish T3 pro-survival action.CONCLUSIONS: THs influence cell survival of ovarian granulosa cells. This effect likely contributes to the TH-induced follicle volume increase.

U2 - 10.1007/s40618-018-0912-2

DO - 10.1007/s40618-018-0912-2

M3 - Article

C2 - 29934772

JO - Journal of Endocrinological Investigation

JF - Journal of Endocrinological Investigation

SN - 0391-4097

ER -