Insulin secretion from beta-cells is affected by deletion of nicotinamide nucleotide transhydrogenase.

• Nicotinamide nucleotide transhydrogenase (NNT) is an inner mitochondrial membrane transmembrane protein involved in regenerating NADPH, coupled with proton translocation across the inner membrane. We have shown that a defect in Nnt function in the mouse, and specifically within the beta-cell, leads to a reduction in insulin secretion. This chapter describes methods for examining Nnt function in the mouse. This includes generating in vivo models with point mutations and expression of Nnt by transgenesis, and making in vitro models, by silencing of gene expression. In addition, techniques are described to measure insulin secretion, calcium and hydrogen peroxide concentrations, membrane potential, and NNT activity. These approaches and techniques can also be applied to other genes of interest.

Subject headings

NATURVETENSKAP  -- Kemi (hsv//swe)

NATURAL SCIENCES  -- Chemical Sciences (hsv//eng)

Keyword

Animals

Calcium

analysis

Cell Line

Gene Silencing

Hydrogen Peroxide

analysis

Insulin

secretion

Insulin-Secreting Cells

enzymology

secretion

Membrane Potential

Mitochondrial

Mice

Mice

Inbred C3H

Mice

Inbred C57BL

Mice

Transgenic

Microscopy

Confocal

Mitochondria

enzymology

genetics

NADP Transhydrogenase

genetics

metabolism

Point Mutation

Publication and Content Type

ref (subject category)

art (subject category)