Intracellular pH (pHin) and cytosolic calcium ([Ca2+]cyt) regulation via ATPases: studies in cell populations, single cells, and subcellular compartments

Jose D. Rojas; Shankar C. Sanka; Sandor Gyorke; Donald E. Wesson; Akwasi Minta; Raul Martinez-Zaguilan

doi:10.1117/12.351011

2 July 1999 Intracellular pH (pHⁱⁿ) and cytosolic calcium ([Ca²⁺]^cyt) regulation via ATPases: studies in cell populations, single cells, and subcellular compartments

Jose D. Rojas, Shankar C. Sanka, Sandor Gyorke, Donald E. Wesson, Akwasi Minta, Raul Martinez-Zaguilan

Author Affiliations +

Proceedings Volume 3600, Biomedical Imaging: Reporters, Dyes, and Instrumentation; (1999) https://doi.org/10.1117/12.351011
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

Changes in pHⁱⁿ and (Ca²⁺)^cyt are important in the signal transduction mechanisms leading to many physiological responses including cell growth, motility, secretion/exocytosis, etc. The concentrations of these ions are regulated via primary and secondary ion transporting mechanisms. In diabetes, specific pH and Ca²⁺ regulatory mechanism might be altered. To study these ions, we employ fluorescence spectroscopy, and cell imagin spectroscopy/confocal microscopy. pH and Ca²⁺ indicators are loaded in the cytosol with acetoxymethyl ester forms of dyes, and in endosomal/lysosomal (E/L) compartments by overnight incubation of cells with dextran- conjugated ion fluorescent probes. We focus on specific pH and Ca²⁺ regulatory systems: plasmalemmal vacuolar- type H⁺-ATPases (pm V-ATPases) and sarcoplasmic/endoplasmic reticulum Ca²⁺-ATPases (SERCA). As experimental models, we employ vascular smooth muscle (VSM) and microvascular endothelial cells. We have chosen these cells because they are important in blood flow regulation and in angiogenesis. These processes are altered in diabetes. In many cell types, ion transport processes are dependent on metabolism of glucose for maximal activity. Our main findings are: (a) glycolysis coupling the activity of SERCA is required for cytosolic Ca²⁺ homeostasis in both VSM and microvascular endothelial cells; (b) E/L compartments are important for pH and Ca²⁺ regulation via H⁺-ATPases and SERCA, respectively; and (c) pm-V- ATPases are important for pHⁱⁿ regulation in microvascular endothelial cells.

Citation Download Citation

Jose D. Rojas, Shankar C. Sanka, Sandor Gyorke, Donald E. Wesson, Akwasi Minta, and Raul Martinez-Zaguilan "Intracellular pH (pHⁱⁿ) and cytosolic calcium ([Ca²⁺]^cyt) regulation via ATPases: studies in cell populations, single cells, and subcellular compartments", Proc. SPIE 3600, Biomedical Imaging: Reporters, Dyes, and Instrumentation, (2 July 1999); https://doi.org/10.1117/12.351011

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available