Evaluation of impaired beta-cell function in nonobese-diabetic (NOD) mouse model using bioluminescence imaging.
Insulin-producing pancreatic β cells are functionally impaired or destroyed in diabetes mellitus. The onset of type 1 diabetes (T1D) represents the culmination of a prolonged prediabetic phase of immune-mediated β-cell destruction. To assess the in vivo metabolic status of these cells, we used the ATP-sensitive firefly luciferase bioluminescence imaging approach, as a noninvasive probe to monitor pathological alterations in β-cell function in the nonobese-diabetic (NOD) mouse model of T1D. Hence, we generated the ToIβ-NOD transgenic mice in which doxycycline-inducible luciferase gene is selectively expressed in β cells. A sharp reduction in bioluminescence emitted in vivo from β cells at the early stages, preceded by several weeks of a limited reduction in β-cell mass. Since this decline could be due to the ongoing inflammatory process occurring in vivo, we exposed control islets to inflammatory cytokines and observed a dramatic decrease in luciferase luminescence, which appears to be due in part to a decrease in protein levels and a drop in intracellular ATP levels. This is the first evidence that selective expression of the luciferase gene represents a sensitive method for noninvasive in vivo monitoring of early β-cell dysfunction, subtle metabolic changes, such as endogenous ATP levels, indicative of a pathological condition in a tissue at the cellular level.
Department of Endocrinology, Hadassah University Hospital, P.O. Box 12000, 91120 Jerusalem, Israel.
Dror
D
Sever
Roy
R
Eldor
Gadi
G
Sadoun
Livnat
L
Amior
Daniele
D
Dubois
Christian
C
Boitard
Claude
C
Aflalo
Danielle
D
Melloul
D000375:Aging; D000818:Animals; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D016688:Mice, Inbred NOD; D008822:Mice, Transgenic; D013997:Time Factors
D007328:Insulin; D008156:Luciferases; D005947:Glucose