Increase of intracellular Ca2+ by purinergic receptors in cultured rat lacrimal gland myoepithelial cells.

ABSTRACT:

Purpose

To isolate and characterize cultured myoepithelial cells (MECs) from rat lacrimal gland and determine which purinergic receptor subtypes are present and functional in MECs.

Methods

Rat lacrimal glands were subjected to collagenase digestion, and MECs were grown. RT-PCR was performed for the purinergic receptors P2X(7), P2Y(1), P2Y(11), and P2Y(13) on RNA isolated from the MECs. Immunofluorescence experiments were performed with antibodies against MEC markers and P2X(7), P2Y(1), P2Y(11), and P2Y(13) purinergic receptors. Proteins from MECs were separated using Western blot analysis techniques. In addition, cells were incubated with Fura 2 tetra acetoxymethyl ester, and intracellular [Ca(2+)] ([Ca(2+)](i)) was determined in response to P2 purinergic agonists.

Result

SUBMITTER: Ohtomo K 

PROVIDER: S-EPMC3341118 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature