Analyzing Ca(2+) dynamics in intact epithelial cells using spatially limited flash photolysis.
Cold Spring Harb Protoc
; 2013(1)2013 Jan 01.
Article
em En
| MEDLINE
| ID: mdl-23282645
ABSTRACT
The production of saliva by parotid acinar cells is stimulated by Ca(2+) activation of Cl(-) and K(+) channels located in the apical plasma membrane of these polarized cells. Here we describe a paradigm for the focal photorelease of either Ca(2+) or an inositol 1,4,5 trisphosphate (InsP(3)) analog. The protocol is designed to be useful for investigating subcellular Ca(2+) dynamics in polarized cells with minimal experimental intervention. Parotid acinar cells are loaded with cell-permeable versions of the caged precursors (NP-EGTA-AM or Ci-InsP(3)/PM). Photolysis is accomplished using a spatially limited, focused diode laser, but the experiment can be readily modified to whole-field photolysis using a xenon flash lamp.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Fotólise
/
Inositol 1,4,5-Trifosfato
/
Cálcio
/
Técnicas Citológicas
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Células Acinares
/
Canais Iônicos
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article