Use of protein kinase C-depleted cells for investigation of the role of protein kinase C in stimulus-response coupling in the pituitary

Craig A. McArdle, P. Michael Conn

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

This chapter describes a method for the preparation of pituitary cell cultures that do not contain measurable protein kinase C activity, or specific phorbol-ester binding sites, and do not respond to protein kinase C-activating phorbol esters. These cells are described as “protein kinase C depleted” because an actual loss of this enzyme provides the most likely explanation for these the effects of phorbol 12-myristate 13-acetate (PMA) pretreatment and for similar effects produced in several other cell types. Although the observations with this model do not exclude a role for protein kinase C in gonadotropin- releasing hormone (GnRH) action, they do suggest that the activation of protein kinase C by GnRH is not required for GnRH-stimulated luteinizing hormone (LH) release or for GnRH-induced homologous desensitization or receptor down-regulation. This chapter also presents an example of the manner in which such cells can be used for assessing whether hormonal activation of this enzyme is required for the mediation of other effects of the hormone.

Original languageEnglish
Pages (from-to)287-301
Number of pages15
JournalMethods in Enzymology
Volume168
Issue numberC
DOIs
StatePublished - 1 Jan 1989
Externally publishedYes

Fingerprint

Dive into the research topics of 'Use of protein kinase C-depleted cells for investigation of the role of protein kinase C in stimulus-response coupling in the pituitary'. Together they form a unique fingerprint.

Cite this