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

by Mark A. Kukucka and Hara P. Misra
Department of Biomedical Sciences
Virginia-Maryland Regional College of Veterinary Medicine
Virginia Polytechnic Institute & 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.


CITATION
Kukucka, Mark A. and Hara P. Misra. 1992. HPLC determination of an
oxytocin-like peptide produced by isolated guinea pig Leydig cells:
stimulation by ascorbate. Arch Androl 29(2):185-90.

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