Dear fellow olfactory scientists and/or electrophysiologists,
For one of my projects, I would like to stimulate/activate different primary olfactory transduction cascade stations. As a first attempt, I wanted to use a mixture of IBMX and forskolin for ACIII/PDE2 since it's a well-established paradigm.
I want to record the transduction current using whole-cell patch clamp recordings in MOE slices of juvenile mice (p3-p8). The preparations and patching work fine; the cells look well and healthy and still have cilia. I keep the cells at -75 mV to -80 mV and stimulate them while recording the membrane current continuously (=voltage-clamp mode).
Unfortunately, so far, I was not able to record any responses. No matter the stimulation paradigm, I only get flatlines and no sign of any current at all. I tried various stimulation concentrations & lengths, but so far, I was not successful. I also tried a Ca2+ imaging approach, and there I got clear responses, so I can rule out the reagents not working. I always use an elevated potassium solution to check for cell targeting and viability in my perfusion system. Next, I wanted to try loose-patch/cell-attached recordings as an intermediate step. I also already checked my filter settings, etc., and I cannot find my mistake!
I would be thankful for any advice or ideas you might have!
Best,
Victoria
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- DNA