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HPLC determination of an oxytocin-like peptide produced by isolated

HPLC determination of an oxytocin-like peptide produced by isolated - Cell Biology and Cell Culture

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Old 11-29-2008, 04:01 PM
tocopherol
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Default 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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