Jump to main content
Jump to site search


A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and in situ affinity constant in cells

Author affiliations

Abstract

Class B G protein-coupled receptors (GPCRs) are involved in a variety of human pathophysiological states. These groups of membrane receptors are less studied than class A GPCRs due to the lack of structural information, delayed small molecule drug discovery, and scarce fluorescence detection tools available. The class B corticotropin-releasing hormone type 1 receptor (CRHR1) is a key player in the stress response whose dysregulation is critically involved in stress-related disorders: psychiatric conditions (i.e. depression, anxiety, and addictions), neuroendocrinological alterations, and neurodegenerative diseases. Here, we present a strategy to label GPCRs with a small fluorescent antagonist that permits the observation of the receptor in live cells through stochastic optical reconstruction microscopy (STORM) with 23 nm resolution. The marker, an aza-BODIPY derivative, was designed based on computational docking studies, then synthesized, and finally tested in biological cells. Experiments on hippocampal neurons demonstrate antagonist effects in similar concentrations as the well-established antagonist CP-376395. A quantitative analysis of two color STORM images enabled the determination of the binding affinity of the new marker in the cellular environment.

Graphical abstract: A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and in situ affinity constant in cells

Back to tab navigation

Supplementary files

Publication details

The article was received on 04 Oct 2018, accepted on 01 Nov 2018 and first published on 01 Nov 2018


Article type: Paper
DOI: 10.1039/C8CP06196C
Citation: Phys. Chem. Chem. Phys., 2018, Advance Article
  •   Request permissions

    A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and in situ affinity constant in cells

    A. M. Szalai, N. G. Armando, F. M. Barabas, F. D. Stefani, L. Giordano, S. E. Bari, C. N. Cavasotto, S. Silberstein and P. F. Aramendía, Phys. Chem. Chem. Phys., 2018, Advance Article , DOI: 10.1039/C8CP06196C

Search articles by author

Spotlight

Advertisements