Stimulatory effect of lactate on testosterone production by rat leydig cells

Christopher Niyibizi, Patrick Smith, Zhibao Mi, Charlotte L. Phillips, Paul Robbins

Research output: Contribution to journalArticle

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Abstract

Previously we found that the increased plasma testosterone levels in male rats during exercise partially resulted from a direct and luteinizing hormone (LH)-independent stimulatory effect of lactate on the secretion of testosterone. In the present study, the acute and direct effects of lactate on testosterone production by rat Leydig cells were investigated. Leydig cells from rats were purified by Percoll density gradient centrifugation subsequent to enzymatic isolation of testicular interstitial cells. Purified rat Leydig cells (1 × 105 cells/ml) were in vitro incubated with human chorionic gonadotropin (hCG, 0.05 IU/ml), forskolin (an adenylyl cyclase activator, 10-5 M), or 8-bromo-adenosine-3′:5′-cyclic monophosphate (8-Br-cAMP, 10-4 M), SQ22536 (an adenylyl cyclase inhibitor, 10-6-10-5 M), steroidogenic precursors (25-hydroxy-cholesterol, pregnenolone, progesterone, and androstenedione, 10-5 M each), nifedipine (a L-type Ca2+ channel blocker, 10-5-10-4 M), or nimodipine (a potent L-type Ca2+ channel antagonist, 10-5-10-4 M) in the presence or absence of lactate at 34°C for 1 h. The concentration of medium testosterone was measured by radioimmunoassay. Administration of lactate at 5-20 mM dose-dependently increased the basal testosterone production by 63-187% but did not alter forskolin- and 8-Br-cAMP-stimulated testosterone release in rat Leydig cells. Lactate at 10 mM enhanced the stimulation of testosterone production induced by 25-hydroxy-cholesterol in rat Leydig cells but not other steroidogenic precursors. Lactate (10 mM) affected neither 30- nor 60-min expressions of cytochrome P450 side chain cleavage enzyme (P450scc) and steroidogenic acute regulatory (StAR) protein. The lactate-stimulated testosterone production was decreased by administration of nifedipine or nimodipine. These results suggested that the physiological level of lactate stimulated testosterone production in rat Leydig cells through a mechanism involving the increased activities of adenylyl cyclase, cytochrome P450scc, and L-type Ca2+ channel.

Original languageEnglish (US)
Pages (from-to)147-154
Number of pages8
JournalJournal of cellular biochemistry
Volume83
Issue number1
DOIs
StatePublished - Sep 22 2001

Fingerprint

Leydig Cells
Testosterone
Rats
Lactic Acid
Adenylyl Cyclases
Nimodipine
Colforsin
Nifedipine
Cytochrome P-450 Enzyme System
Cholesterol
Pregnenolone
Density Gradient Centrifugation
Centrifugation
Androstenedione
Chorionic Gonadotropin
Enzymes
Luteinizing Hormone
Adenosine
Radioimmunoassay
Progesterone

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Niyibizi, Christopher ; Smith, Patrick ; Mi, Zhibao ; Phillips, Charlotte L. ; Robbins, Paul. / Stimulatory effect of lactate on testosterone production by rat leydig cells. In: Journal of cellular biochemistry. 2001 ; Vol. 83, No. 1. pp. 147-154.
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Stimulatory effect of lactate on testosterone production by rat leydig cells. / Niyibizi, Christopher; Smith, Patrick; Mi, Zhibao; Phillips, Charlotte L.; Robbins, Paul.

In: Journal of cellular biochemistry, Vol. 83, No. 1, 22.09.2001, p. 147-154.

Research output: Contribution to journalArticle

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N2 - Previously we found that the increased plasma testosterone levels in male rats during exercise partially resulted from a direct and luteinizing hormone (LH)-independent stimulatory effect of lactate on the secretion of testosterone. In the present study, the acute and direct effects of lactate on testosterone production by rat Leydig cells were investigated. Leydig cells from rats were purified by Percoll density gradient centrifugation subsequent to enzymatic isolation of testicular interstitial cells. Purified rat Leydig cells (1 × 105 cells/ml) were in vitro incubated with human chorionic gonadotropin (hCG, 0.05 IU/ml), forskolin (an adenylyl cyclase activator, 10-5 M), or 8-bromo-adenosine-3′:5′-cyclic monophosphate (8-Br-cAMP, 10-4 M), SQ22536 (an adenylyl cyclase inhibitor, 10-6-10-5 M), steroidogenic precursors (25-hydroxy-cholesterol, pregnenolone, progesterone, and androstenedione, 10-5 M each), nifedipine (a L-type Ca2+ channel blocker, 10-5-10-4 M), or nimodipine (a potent L-type Ca2+ channel antagonist, 10-5-10-4 M) in the presence or absence of lactate at 34°C for 1 h. The concentration of medium testosterone was measured by radioimmunoassay. Administration of lactate at 5-20 mM dose-dependently increased the basal testosterone production by 63-187% but did not alter forskolin- and 8-Br-cAMP-stimulated testosterone release in rat Leydig cells. Lactate at 10 mM enhanced the stimulation of testosterone production induced by 25-hydroxy-cholesterol in rat Leydig cells but not other steroidogenic precursors. Lactate (10 mM) affected neither 30- nor 60-min expressions of cytochrome P450 side chain cleavage enzyme (P450scc) and steroidogenic acute regulatory (StAR) protein. The lactate-stimulated testosterone production was decreased by administration of nifedipine or nimodipine. These results suggested that the physiological level of lactate stimulated testosterone production in rat Leydig cells through a mechanism involving the increased activities of adenylyl cyclase, cytochrome P450scc, and L-type Ca2+ channel.

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