Functional identification and monitoring of individual alpha and beta cells in cultured mouse islets of Langerhans
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Functional identification and monitoring of individual alpha and beta cells in cultured mouse islets of Langerhans. / Hjortoe, G M; Hagel, G M; Terry, B R; Thastrup, Ole; Arkhammar, P O G.
In: Acta Diabetologica, Vol. 41, No. 4, 2004, p. 185-93.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Functional identification and monitoring of individual alpha and beta cells in cultured mouse islets of Langerhans
AU - Hjortoe, G M
AU - Hagel, G M
AU - Terry, B R
AU - Thastrup, Ole
AU - Arkhammar, P O G
PY - 2004
Y1 - 2004
N2 - The aim of the present study was to evaluate, by use of fluorescence microscopy and immunofluorescence stainings, the use of a fluorescent membrane potential sensitive probe as a means to identify and monitor changes in membrane potential of individual cell types in whole islets of Langerhans over time. Our work supports the use of the fluorescent probe bis-(1,3 dibutylbarbituric acid) trimethine oxonol (diBAC(4)(3)), in identification of single alpha and beta cells in the periphery of mouse pancreatic islets cultured on extracellular matrix. At a low extracellular glucose concentration (3 mM), heterogeneous staining of the islets was observed. Approximately 97% of the peripheral cells that stained brightly with diBAC(4)(3) were glucagon positive. Additional diBAC(4)(3) studies, demonstrated that an increase in glucose concentration from 3 to 10 mM is paralleled by repolarization of alpha cells and depolarization of beta cells. This suggests that reciprocity of glucagon and insulin release upon glucose stimulation is coupled to divergent changes in membrane potential of these cell types and supports the use of diBAC(4)(3) as a means to detect changes in secretion in both cell types.
AB - The aim of the present study was to evaluate, by use of fluorescence microscopy and immunofluorescence stainings, the use of a fluorescent membrane potential sensitive probe as a means to identify and monitor changes in membrane potential of individual cell types in whole islets of Langerhans over time. Our work supports the use of the fluorescent probe bis-(1,3 dibutylbarbituric acid) trimethine oxonol (diBAC(4)(3)), in identification of single alpha and beta cells in the periphery of mouse pancreatic islets cultured on extracellular matrix. At a low extracellular glucose concentration (3 mM), heterogeneous staining of the islets was observed. Approximately 97% of the peripheral cells that stained brightly with diBAC(4)(3) were glucagon positive. Additional diBAC(4)(3) studies, demonstrated that an increase in glucose concentration from 3 to 10 mM is paralleled by repolarization of alpha cells and depolarization of beta cells. This suggests that reciprocity of glucagon and insulin release upon glucose stimulation is coupled to divergent changes in membrane potential of these cell types and supports the use of diBAC(4)(3) as a means to detect changes in secretion in both cell types.
KW - Animals
KW - Arginine
KW - Barbiturates
KW - Dose-Response Relationship, Drug
KW - Female
KW - Fluorescent Antibody Technique
KW - Fluorescent Dyes
KW - Glucagon
KW - Glucose
KW - Islets of Langerhans
KW - Isoxazoles
KW - Leucine
KW - Membrane Potentials
KW - Mice
KW - Mice, Inbred Strains
KW - Microscopy, Fluorescence
KW - Osmolar Concentration
KW - Staining and Labeling
KW - Time Factors
KW - Tissue Culture Techniques
U2 - 10.1007/s00592-004-0164-9
DO - 10.1007/s00592-004-0164-9
M3 - Journal article
C2 - 15660202
VL - 41
SP - 185
EP - 193
JO - Acta Diabetologica
JF - Acta Diabetologica
SN - 0940-5429
IS - 4
ER -
ID: 43348979