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

doi:10.1111/j.1365-2605.1982.tb00235.x

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

^*Corresponding author for this work

University of Manitoba

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

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 language	English
Pages (from-to)	74-80
Number of pages	7
Journal	International Journal of Andrology
Volume	5
Issue number	1
DOIs	https://doi.org/10.1111/j.1365-2605.1982.tb00235.x
State	Published - Feb 1982
Externally published	Yes

Keywords

3β‐HSD
isolated Leydig cells

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10.1111/j.1365-2605.1982.tb00235.x

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title = "Determination of Δ5 3β‐hydroxysteroid dehydrogenase activity in intact isolated rat Leydig cells",

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.",

keywords = "3β‐HSD, isolated Leydig cells",

author = "Paz, {G. F.} and Winter, {J. S.D.} and Reyes, {F. T.} and C. Faiman",

year = "1982",

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doi = "10.1111/j.1365-2605.1982.tb00235.x",

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AU - Winter, J. S.D.

AU - Reyes, F. T.

AU - Faiman, C.

PY - 1982/2

Y1 - 1982/2

N2 - 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.

AB - 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.

KW - 3β‐HSD

KW - isolated Leydig cells

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

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Keywords

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