3D-reconstruction of connexin32 channels distribution in the myelin of Schwann cells by advanced optical microscopy

Connexin 32 (Cx32) is a 32 kDa protein of the connexin family that is expressed in the peripheral nervous system where it localizes in the myelin sheath of Schwann cells. Mutations of Cx32 are the leading cause of the X-linked form of Charcot–Marie–Tooth disease (CMT1X), a peripheral neuropathy for which there is no cure. Alteration in the distribution and function of Cx32 channels are presumed to trigger the neuropathy, but the pathological mechanism is still unknown. In this thesis work we combined two-photon fluorescence microscopy with third harmonic generation of the myelin sheath of Schwann cells to analyze the distribution of Cx32 and 3D render it by a software we developed in Matlab. STED microscopy tests were carried out with the future perspective to obtain high-resolution 3D images of Cx32 distribution in nerve samples of CMT1X patients.