Nuclear and Membrane Receptors for Sex Steroids Are Involved in the Regulation of Delta/Serrate/LAG-2 Proteins in Rodent Sertoli Cells.

Sylwia Lustofin,Barbara Bilińska,Joanna Cyran,Anna Hejmej,Małgorzata Brzoskwinia,Alicja Kamińska,Małgorzata Kotula-Balak

doi:10.3390/ijms23042284

Sylwia Lustofin, Barbara Bilińska + Show 5 more

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https://doi.org/10.3390/ijms23042284

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Abstract

Delta/Serrate/LAG-2 (DSL) proteins, which serve as ligands for Notch receptors, mediate direct cell–cell interactions involved in the determination of cell fate and functioning. The present study aimed to explore the role of androgens and estrogens, and their receptors in the regulation of DSL proteins in Sertoli cells. To this end, primary rat Sertoli cells and TM4 Sertoli cell line were treated with either testosterone or 17β-estradiol and antagonists of their receptors. To confirm the role of particular receptors, knockdown experiments were performed. mRNA and protein expressions of Jagged1 (JAG1), Delta-like1 (DLL1), and Delta-like4 (DLL4) were analyzed using RT-qPCR, Western blot, and immunofluorescence. Testosterone caused downregulation of JAG1 and DLL1 expression, acting through membrane androgen receptor ZRT- and Irt-like protein 9 (ZIP9) or nuclear androgen receptor (AR), respectively. DLL4 was stimulated by testosterone in the manner independent of AR and ZIP9 in Sertoli cells. The expression of all studied DSL proteins was upregulated by 17β-estradiol. Estrogen action on JAG1 and DLL1 was mediated chiefly via estrogen receptor α (ERα), while DLL4 was controlled via estrogen receptor β (ERβ) and membrane G-protein-coupled estrogen receptor (GPER). To summarize, the co-operation of nuclear and membrane receptors for sex steroids controls DSL proteins in Sertoli cells, contributing to balanced Notch signaling activity in seminiferous epithelium.

Highlights

A series of studies performed at the beginning of the 21st century and subsequent research clearly demonstrated that proper androgen/estrogen balance is fundamental for normal male sexual development and function in humans and animals [1,2,3,4,5,6]
To further confirm the mechanisms involved in JAG1 regulation by testosterone, androgen receptor (AR) and ZRT- and Irt-like protein 9 (ZIP9) were knocked down in Sertoli cells
We have demonstrated the role of sex steroids and their respective receptors in the control of DSL proteins in rodent Sertoli cells

Summary

Introduction

A series of studies performed at the beginning of the 21st century and subsequent research clearly demonstrated that proper androgen/estrogen balance is fundamental for normal male sexual development and function in humans and animals [1,2,3,4,5,6]. This balance is governed primarily by aromatase, which catalyzes the irreversible conversion of androgenic steroids (testosterone and androstenedione), produced by testicular interstitium, into the estrogens (estradiol and estrone), as well as by the expression of nuclear androgen (AR). Rodent postnatal Sertoli cells show a dynamic pattern of the expression of nuclear and membrane androgen and estrogen receptors, thereby enabling a co-ordinated response to hormonal stimulation [12,19,21,22]

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