HPLC determination of an oxytocin-like peptide produced by isolated

HPLC determination of an oxytocin-like peptide produced by isolated
guinea pig Leydig cells: stimulation by ascorbate.
Arch Androl. 1992 Sep-Oct;29(2):185-90.

Kukucka, Mark A. and Misra, Hara P.

Department of Biomedical Sciences, Virginia-Maryland Regional College
of Veterinary Medicine, Virginia Polytechnic Institute and State
University, Blacksburg 24061.

ABSTRACT

Highly purified populations of guinea pig Leydig cells were incubated
with a maximally stimulating dose of 100 ng/mL LH for 24 h in the
presence of increasing concentrations of sodium ascorbate. Sample
supernatants were extracted, concentrated under vacuum, and
reconstituted with acidified absolute ethanol. Samples were analyzed
for oxytocin using high-performance liquid chromatography with
electrochemical detection and known concentrations of an authentic
oxytocin standard. Leydig cells stimulated with 0, 25, and 50 microM
ascorbate produced and secreted 40.1 +/- 1.23, 77.4 +/- 13.8, 74.2 +/-
26.3 pg of an oxytocin-like peptide, respectively, per 1 x 10(6)
cells. These results indicate that guinea pig Leydig cells are capable
of producing an oxytocin-like peptide de novo and that low
concentrations of ascorbate stimulate the production of this peptide
in Leydig cells cultured in vitro.

On Mar 21, 3:19*pm, tocopherol <[Only registered users see links. ]> wrote:

Published in: Systems Biology in Reproductive Medicine, Volume 29,
Issue 2 September 1992 , pages 185 - 190

Previously published as: Archives of Andrology (0148-5016, 1521-0375)
until 2008


HPLC Determination of an Oxytocin-Like Peptide Produced by Isolated
Guinea Pig Leydig Cells: Stimulation by Ascorbate

by Mark A. Kukucka and Hara. P. Misra

Affiliation: Department of Biomedical Sciences, Virginia-Maryland
Regional College of Veterinary Medicine, Virginia Polytechnic
Institute and State University, Blacksburg, VA 24061

ABSTRACT

Highly purified populations of guinea pig Leydig cells were incubated
with a maximally stimulating dose of 100 ng/mL LH for 24 h in the
presence of increasing concentrations of sodium ascorbate. Sample
supernatants were extracted, concentrated under vacuum, and
reconstituted with acidified absolute ethanol. Samples were analyzed
for oxytocin using high-performance liquid chromatography with
electrochemical detection and known concentrations of an authentic
oxytocin standard. Leydig cells stimulated with 0, 25, and 50 M
ascorbate produced and secreted 40.1 ± 1.23, 77.4 ± 13.8, 74.2 ± 26.3
pg of an oxytocin-like peptide, respectively, per 1 ± 106 cells. These
results indicate that guinea pig Leydig cells are capable of producing
an oxytocin-like peptide de novo and that low concentrations of
ascorbate stimulate the production of this peptide in Leydig cells
cultured in vitro.

Keywords: Oxytocin; Leydig cells; Ascorbate; HPLC; Testis; Sodium
ascorbate; Peptide

DOI: 10.3109/01485019208987723