Abstract
Platelet-derived growth factor receptor α (PDGFRα)-positive peritubular cells (PTCs) are suggested to be putative stem Leydig cells. At present little is known about their phenotype and steroidogenic potential. We isolated highly purified PDGFRα-positive neonatal PTCs by magnetic cell sorting (MACS) from 8dpp rat testes and characterized them in vitro. We have demonstrated that PDGFRα-positive PTCs have a mixed phenotype. They expressed PTC-specific genes (αSma, Myh11), pluripotency markers (Pou5f1, nestin, Lifr) and genes encoding steroidogenic enzymes. Treatment with the cAMP-analog (Bu)2cAMP for 7 days upregulated steroidogenic enzyme gene expression and significantly increased their steroidogenic potential. The main end-point steroid was progesterone due to rapid inactivation of CYP17 and 17βHSD. Long-term culturing of PDGFRα-positive PTCs increased the expression of Myh11, and treatment with (Bu)2cAMP attenuated this process. All together, our findings support the hypothesis that neonatal PDGFRα-positive PTCs are steroidogenic competent progeny of stem Leydig cells (SLCs) which give rise to the adult Leydig cell lineage.
Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Animals, Newborn
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Cell Differentiation
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Cholesterol Side-Chain Cleavage Enzyme / genetics
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Cholesterol Side-Chain Cleavage Enzyme / metabolism
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Cyclic AMP / metabolism
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Gonadal Steroid Hormones / biosynthesis
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Intermediate Filament Proteins / metabolism
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Leukemia Inhibitory Factor Receptor alpha Subunit / metabolism
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Leydig Cells / enzymology*
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Leydig Cells / metabolism
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Male
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Multienzyme Complexes / genetics
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Multienzyme Complexes / metabolism
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Nerve Tissue Proteins / metabolism
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Nestin
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Phenotype
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Primary Cell Culture
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Progesterone / biosynthesis*
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Progesterone Reductase / genetics
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Progesterone Reductase / metabolism
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Rats
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Rats, Sprague-Dawley
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Receptor, Platelet-Derived Growth Factor alpha / metabolism*
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Second Messenger Systems
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Seminiferous Tubules / cytology
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Stem Cells / physiology
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Steroid 17-alpha-Hydroxylase / genetics
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Steroid 17-alpha-Hydroxylase / metabolism
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Steroid Isomerases / genetics
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Steroid Isomerases / metabolism
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Testosterone / biosynthesis*
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Transcriptome
Substances
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3 beta-hydroxysteroid oxidoreductase-delta(5) 3-ketosteroid isomerase
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Gonadal Steroid Hormones
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Intermediate Filament Proteins
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Leukemia Inhibitory Factor Receptor alpha Subunit
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Lifr protein, rat
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Multienzyme Complexes
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Nerve Tissue Proteins
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Nes protein, rat
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Nestin
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Testosterone
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Progesterone
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Cyclic AMP
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Progesterone Reductase
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CYP17A1 protein, rat
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Steroid 17-alpha-Hydroxylase
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Cholesterol Side-Chain Cleavage Enzyme
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Receptor, Platelet-Derived Growth Factor alpha
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Steroid Isomerases