Ca2+ homeostasis in microvascular endothelial cells from an insulin dependent diabetic model: role of endosomes/Lysosomes

Shankar C. Sanka, David C. Bennett, Jose D. Rojas, Geraldine B. Tasby, Cynthia J. Meininger, Guoyao Wu, Donald E. Wesson, Curt Pfarr, Raul Martinez-Zaguilan

Research output: Contribution to journalConference article

1 Scopus citations

Abstract

Cytosolic Ca2+ ([Ca2+]cyt) regulates several functions, e.g. cell growth, contraction, secretion, etc. In many cell types, ion homeostasis appears to be coupled with glucose metabolism. In certain cell types, a strict coupling between glycolysis and the activity of Sarcoplasmic/Endoplasmic Reticulum Ca2+-ATPases (SERCA) has been suggested. Glucose metabolism is altered in diabetes. We hypothesize that: (a) Ca2+ homeostasis is altered in microvascular endothelial cells from diabetic animals due to the dysfunction of glycolysis coupling the activity of SERCA; (b) endosomal/lysosomal (E/L) compartments expressing SERCA are involved in the dysfunction associated with diabetes. Ca2+ ions in E/L compartments can be studied by either spectral imaging/confocal microscopy in single cells, or by fluorescence spectroscopy in cell populations. In cell populations, agonist stimulation elicited greater [Ca2+]cyt increases in cells from diabetic than from normal animals. Simultaneous measurements of [Ca2+]cyt and Ca2+ in E/L compartments ([Ca2+]E/L) using fluorescence spectroscopy and spectral imaging/confocal microscopy, demonstrate that Ca2+ is released from E/L compartments following agonist stimulation. Immunocytochemical studies demonstrate that E/L compartments exhibit the machinery to regulate Ca2+: SERCA and ryanodine receptor (RyR) coupled Ca2+ channels. Our functional studies, using confocal/spectral imaging microscopy, indicated that E/L compartments employ SERCA and RyR coupled Ca2+ channels to refill and release Ca2+, respectively. Glucose modulates refilling of E/L compartments with Ca2+ via SERCA, since SERCA inhibitors suppress this refilling. The regulation of these phenomena are altered in diabetes. These data indicate that E/L compartments are important for Ca2+ homeostasis.

Original languageEnglish (US)
Pages (from-to)56-66
Number of pages11
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume3924
StatePublished - Jan 1 2000
Externally publishedYes
EventMolecular Imaging: Reporters, Dyes, Markers, and Instrumentation - San Jose, CA, USA
Duration: Jan 23 2000Jan 24 2000

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ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

Cite this

Sanka, S. C., Bennett, D. C., Rojas, J. D., Tasby, G. B., Meininger, C. J., Wu, G., Wesson, D. E., Pfarr, C., & Martinez-Zaguilan, R. (2000). Ca2+ homeostasis in microvascular endothelial cells from an insulin dependent diabetic model: role of endosomes/Lysosomes. Proceedings of SPIE - The International Society for Optical Engineering, 3924, 56-66.