PUBLICATION
Incorporating phototransduction proteins in zebrafish green cone with pressure-polished patch pipettes
- Authors
- Aquila, M., Dell'Orco, D., Fries, R., Koch, K.W., Rispoli, G.
- ID
- ZDB-PUB-190729-20
- Date
- 2019
- Source
- Biophysical chemistry 253: 106230 (Journal)
- Registered Authors
- Keywords
- Calcium signalling, Guanylate cyclase, Phototransduction, Retinal photoreceptors, Zebrafish retina, cGMP
- MeSH Terms
-
- Animals
- Antibodies, Monoclonal/metabolism
- Calcium/metabolism
- Guanylate Cyclase-Activating Proteins/metabolism*
- Light Signal Transduction*
- Pressure
- Retinal Cone Photoreceptor Cells/metabolism*
- Zebrafish
- PubMed
- 31352142 Full text @ Biophys. Chem.
Citation
Aquila, M., Dell'Orco, D., Fries, R., Koch, K.W., Rispoli, G. (2019) Incorporating phototransduction proteins in zebrafish green cone with pressure-polished patch pipettes. Biophysical chemistry. 253:106230.
Abstract
The neuronal Ca2+-sensor guanylate cyclase-activating protein 3 (zGCAP3) is a major regulator of guanylate cyclase (GC) activity expressed in zebrafish cone cells. Here, the zGCAP3, or a monoclonal antibody directed against zGCAP3, was injected in the cone cytoplasm by employing the pressure-polished pipette technique. This technique allows to perform "real time" zGCAP3 (or of any other phototransduction protein) over-expression or knock-down, respectively, via the patch pipette. Photoresponses were not affected by purified zGCAP3, indicating that GC was already saturated with endogenous zGCAP3. The cytosolic injection of anti-zGCAP3 produced the slowing down kinetics of the flash response recovery, as theoretically expected by a minimal phototransduction model considering the antibody acting exclusively on the maximal GC activation by low Ca2+. However, the antibody produced a progressive current decay toward the zero level, as if the antibody affected also the basal GC activity in the dark.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping