A method to determine insulin responsiveness in synaptosomes isolated from frozen brain tissue.

ABSTRACT:

Background

Studying the insulin signaling response at the synapse is an important approach to understand molecular mechanisms involved in disease-related neurodegenerative processes.

New method

We developed a method for studying the insulin responsiveness at the synaptic level by isolating functional synaptosomes from fresh or frozen tissue and exposing them to insulin in the presence of ATP (a critical step) to detect insulin receptor (IR) activation.

Results

We performed an ATP dose-response curve, insulin dose-response curve, and insulin response time course to optimize this method. We also demonstrated that our protocol reflects the degree of insulin responsiveness in vivo by using an animal model of known insulin resistance, AtENPP1-Tg mice.

Comparison with ex

SUBMITTER: Franklin W 

PROVIDER: S-EPMC4749422 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature