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Related Experiment Videos

Estrogen receptor messenger ribonucleic acid changes during Leydig cell development

J Zhai1, K D Lanclos, T O Abney

  • 1Department of Physiology and Endocrinology, Medical College of Georgia, August 30912, USA.

Biology of Reproduction
|October 1, 1996
PubMed
Summary
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Mature Leydig cells (LCs) regenerate from precursor Leydig cells (PLCs) after ethane dimethane sulfonate (EDS) treatment. Precursor Leydig cells (PLCs) have higher estrogen receptor (ER) mRNA levels, which decrease during differentiation into LCs, potentially regulating Leydig cell regeneration.

Area of Science:

  • Reproductive biology
  • Endocrinology
  • Cell biology

Background:

  • Leydig cells (LCs) are crucial for testosterone production in the testes.
  • Precursor Leydig cells (PLCs) are believed to differentiate into mature LCs.
  • The role of estrogen receptors (ERs) in Leydig cell development and function is not fully understood.

Purpose of the Study:

  • To investigate the presence and levels of estrogen receptor (ER) mRNA in precursor Leydig cells (PLCs) and Leydig cells (LCs).
  • To examine the changes in ER mRNA levels and testosterone (T) production during Leydig cell regeneration following ethane dimethane sulfonate (EDS) treatment.
  • To elucidate the potential role of ERs in the differentiation of PLCs into LCs.

Main Methods:

  • Mature male Sprague-Dawley rats were treated with ethane dimethane sulfonate (EDS).

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  • Precursor Leydig cells (PLCs) and Leydig cells (LCs) were isolated using collagenase digestion and Percoll gradient centrifugation.
  • Estrogen receptor (ER) mRNA levels were quantified using reverse transcriptase polymerase chain reaction (RT-PCR).
  • In vitro testosterone (T) production was measured by radioimmunoassay (RIA) in response to hCG stimulation.

    • Main Results:

    • ER mRNA was detected in both PLC and LC fractions.
    • PLC fractions exhibited significantly higher ER mRNA levels (20-fold) compared to LC fractions in control rats.
    • EDS treatment led to a decrease in ER mRNA in PLCs, reaching a nadir at 16 days posttreatment, followed by gradual recovery.
    • Testosterone production in PLCs was initially undetectable after EDS treatment, with hCG responsiveness emerging at 16 days and peaking between 4-6 weeks.
    • LCs showed no detectable testosterone production at 2-10 days post-EDS, with gradual recovery of basal and hCG-stimulated production from 16 days onwards.

    Conclusions:

    • Functional Leydig cells (LCs) regenerate from precursor Leydig cells (PLCs) after EDS-induced injury.
    • Both PLCs and LCs express estrogen receptor (ER) mRNA, with higher levels in PLCs.
    • A decrease in ER mRNA in PLCs coincides with their differentiation into LCs, suggesting ER signaling may regulate this process.
    • Local estradiol-17 beta production by LCs might act via ERs to inhibit PLC development, and a decrease in ERs or estradiol could release PLCs for regeneration.