Mutated ATP synthase induces oxidative stress and impaired insulin secretion in beta-cells of female BHE/cdb rats.
BACKGROUND
Adenosine triphosphate (ATP) is a critical determinant of beta-cell insulin secretion in response to glucose. BHE/cdb rats have a mutation in ATP synthase that limits ATP production, yet develop mild diabetes only with ageing. We investigated the cellular basis for reduced insulin secretion and compensatory mechanisms that mitigate the effects of the ATP synthase mutation.


METHODS
In vitro beta-cell function in isolated islets and expression of key regulatory genes was compared with in vivo oral glucose tolerance and insulin sensitivity in BHE/cdb and control rats.


RESULTS
BHE/cdb rat islets had reduced responsiveness to glucose stimulation and ATP content was 35% lower than in control islets. Oral glucose tolerance was impaired at both 21 and 43 weeks of age because of a reduction in glucose-stimulated insulin secretion (GSIS). An increase in inducible nitric oxide synthase (INOS, 3-fold) and manganese superoxide dismutase (MnSOD, 1.6-fold), detection of nitrotyrosine, beta-cell apoptosis, and nucleocytoplasmic translocation of pancreas duodenum homeobox-1 (PDX-1) in beta-cells indicated increased oxygen radical formation. However, BHE/cdb rats partially compensated for low glucose responsiveness by increasing the number of small islets and beta-cell hypertrophy. There was also an increase in the proportion of mature insulin relative to proinsulin (PI) detected within beta-cell granules. Increased activation of AMP-dependent kinase (AMPK)-regulated pathways was consistent with increased oxidative stress and with induction of apoptosis and reduction of preproinsulin gene transcription.


CONCLUSIONS
The findings are consistent with impaired but partially compensated mechanisms of insulin secretion early in life, but progressive non-compensated impairments due to oxidative stress occurs by age 43 weeks.
Saleh
Monique C
MC
Department of Biomedical Sciences, University of Prince Edward Island, Charlottetown, PE, Canada.
Fatehi-Hassanabad
Zahra
Z
Wang
Rennian
R
Nino-Fong
Rodolpho
R
Wadowska
Dorota W
DW
Wright
Glenda M
GM
Harper
Mary-Ellen
ME
Chan
Catherine B
CB
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D005260:Female; D005951:Glucose Tolerance Test; D007328:Insulin / Q000378:metabolism* / Q000502:physiology; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism*; D007515:Islets of Langerhans / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology*; D010641:Phenotype; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000502:physiology; D011384:Proinsulin / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000235:genetics*; D051381:Rats
D007328:Insulin; D000255:Adenosine Triphosphate; D011384:Proinsulin; C118888:AMP-dependent kinase (ATP-forming); D017856:Phosphotransferases (Phosphate Group Acceptor); D006180:Proton-Translocating ATPases