TRPM4 Immunos Yield Encouraging Results

After fixing the TRPM4 protocol to work with an Alexa 488 flurophore, I was able to take some high quality confocal images of the medullary brainstem. This slide is slightly caudal for the preBotC, but still contains the preBotC on the right portion of the coronal, 60 micron slice. This image shows TRPM4 prevalence in the preBotC as well as near the fourth ventricle.

Screen Shot 2018-07-10 at 4.04.32 PM

The cyan color scheme most accurately resembles the TritC channel for the 488 fluorophore, which shows the medullary slice contains both lateral preBotCs as well as the TRPM4s.

Screen Shot 2018-07-10 at 4.07.57 PM

When using a 40X lens to zoom into the preBotC, the following anatomical data was found.

Screen Shot 2018-07-10 at 4.11.41 PM

While there is some photobleaching of the dye due to high gain settings in the confocal microscope, the TRPM4 channels are clearly prevalent in the preBotC. I am in the process of creating a novel immunoassay to utilize both TRPC3 and TRPM4 antibodies to demonstrate the calcium-dependent nature of respiratory rhythmogenesis in mammalian life.

Speak Your Mind