Development and characterization of in vivo models for Photopharmacology

Abstract

The main aim of this thesis is setting up and using in vivo screening techniques to identify and characterize new photopharmacological compounds targeting endogenous proteins.

In order to achieve that aim, the thesis has been structured in three specific objectives, which are developed in the corresponding parts and chapters indicated below:

Objective I. Photomanipulation of cardiac activity and organism development (part I).

- Establish a procedure to identify photoswitchable compounds affecting zebrafish larvae development based on phenotypic outcomes during embryogenesis and hatching (Chapter 2).

- Identification of relevant morphological, anatomical, and organotypic traits that are affected by the departure compound of photoswitchable drugs (Chapter 2).

- Study of phenotypic markers as a function of photoswitchable drug isoforms (Chapter 2).

- Developing a strategy for real-time recording of early stage tadpoles’ cardiac function under controlled illumination (Chapter 3).

- Developing a computational approach for video extracting readouts and analysis of cardiac parameters (Chapter 3).

- Identification of photoswitchable compounds with reversible effects on tadpoles’ cardiac function (Chapter 3).

Objective II. Photomanipulation of locomotion (part II).

- Establish an experimental and analytical procedure to categorise differences in locomotion outcomes under two wavelengths illumination (Chapter 4).

- Identify swimming profiles related to inhibitory neurotransmission pathways of zebrafish larvae in darkness and illumination (Chapter 4).

- Devise a strategy to screen photoswitchable compounds using zebrafish larvae activity profiles and their alteration under varying light conditions (Chapter 4).

- Identification and characterization of photoswitchable GABAAR drugs using zebrafish locomotion assays (Chapter 5).

- Identify swimming profiles related to inhibitory neural transmission pathways present during locomotion in Xenopus tropicalis tadpoles under long relaxed dark periods and light stimuli (Chapter 6).

- Identification and characterization of a photoswitchable potentiator of glycinergic transmission using tapole swimming assays (Chapter 6).

- Identification and characterization of a photoswitchable ligands of adrenergic receptors using zebrafish swimming assays (Chapter 7).

- Identification and characterization of a photoswitchable ligands of dopaminergic receptors using zebrafish swimming assays (Chapter 8).

Objective III. Photomanipulation of visual function (part III).

- Develop a model of acute blinding in zebrafish larvae by photoinduced retinal degeneration in order to test vision restoration drugs (Chapter 9).

- Identification and characterization of locomotive responses in blinded larvae (Chapter 9).

- Development and validation of an optokinetic visual-motor reflex assay to test vision restoration drugs (Chapter 9).