Insulin resistance induced by long-term hyperinsulinemia abolishes the effects of acute insulin exposure on cell-surface nicotinic acetylcholine receptor levels and actin cytoskeleton morphology

GARCIA, ANA PAULA; GAYDOU, LUISA; PÉREZ, EUGENIA; BARRANTES, FRANCISCO J.

Biochemical and Biophysical Research Communications

Elsevier B.V.

Lugar: Utrecht; Año: 2023 vol. 685

Using CHO–K1/A5 cells, a clonal cell line that robustly expresses adult muscle-type nicotinic acetylcholine receptor (nAChR), we explored whether insulin resistance in these mammalian cells affects cell-surface expression of the nAChR, its endocytic internalization, and actin cytoskeleton integrity. Acute nanomolar insulin stimulation resulted in a slow increase in nAChR cell-surface levels, reaching maximum levels at ~1 h. Long periods of insulin incubation caused CHO–K1/A5 cells to become insulin resistant, as previously observed with several other cell types. Furthermore, long-term insulin treatment abolished the effects of short-term insulin exposure on cell-surface nAChR levels, suggestive of a desensitization phenomenon. It also affected the kinetics of ligand induced nAChR internalization. Since the integrity of the cortical actin cytoskeleton affects nAChR endocytosis, we also studied the effects of long-term insulin treatment on this meshwork. We found that it significantly affected the cortical actin morphology of CHO–K1/A5 cells and the response of the actin cytoskeleton to a subsequent short-term insulin stimulus. Overall, the present results show for the first time the effects of insulin signaling on cell-surface nAChR expression and actin cytoskeleton-associated internalization.