Determination of Δ5 3β‐hydroxysteroid dehydrogenase activity in intact isolated rat Leydig cells

G. F. Paz, J. S.D. Winter, F. T. Reyes, C. Faiman

Research output: Contribution to journalArticlepeer-review

Abstract

A method for the determination of Δ5 3β‐hydroxysteroid dehydrogenaseisomerase (3β‐HSD) activity in intact isolated Leydig cells was established. This method utilizes the conversion of [7‐3H]dehydroepiandrosterone (1.04 μmole) to androstenedione and expresses the activity of the enzyme as μmoles of androstenedione produced/μg DNA/h. The reaction is limited to 0.5‐4 μg DNA of Leydig cells/ml (equivalent to 0.1‐0.8 million of Levdig cells/ml) and to 1 h of incubation at 34°C. The 3β‐HSD activity of 44 suspensions of Leydig cells isolated from adult rats was found to be 1.13 pL 0.03 (SE) μmoles/μg DNA/h. This new method for direct measurement of 3β‐HSD activity in intact Leydig cells was found to be rapid, easy to perform and highly reproducible.

Original languageEnglish
Pages (from-to)74-80
Number of pages7
JournalInternational Journal of Andrology
Volume5
Issue number1
DOIs
StatePublished - Feb 1982
Externally publishedYes

Keywords

  • 3β‐HSD
  • isolated Leydig cells

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