Lazrak-dataset
Description
Electrophysiology data of ENaC activity in human and mouse alveolar epithelial cells in-situ, in freshly cut lung slices, and in culture using patch clamp techniques and Ussing chamber system for measuring short circuit currents across alveolar cells confluents monolayers. In addition, the project will generate lung tissue images for the distribution and the expression of the calcium sensing receptor in the alveolar epithelium. The expression of ENaC and CaSR proteins will be measured by western blot producing more data to help clarify the mechanism by which hyaluronan fragments activate calcium-sensing receptor and inhibits ions and water transport across the alveolar epithelium causing lung edema. The collected data from the experiments mentioned above will approximatively total 20 to 30 Gigabytes. The following data files will be produced in the course of the project: Axon binary files (ABF files) when measuring ENaC activity in alveolar cells in-situ, in lung slices. These files are usually larges, tens of megabytes each, depending on the sampling rate and the time necessary to collect enough data and test compounds of interest. In addition, the data analysis will produce new files, axon text files (ATF files), and finally the processed data will be transferred to a third-party analysis and presentation programs ( IGOR from Wavemetrics and Origin from Origin labs) that generate the final files suitable for presentation and publication. The imaging files are very large and remain at the same size after analysis. Western blot files and in-vivo data will be presented as an aggregate and do not generate large files excel and PowerPoint will be used to pool the numbers together and put in a presentable form, respectively.
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Steps to reproduce
we used patch clamp electrophysiology and Ussing chamber system for studying epithelial sodium channel (ENaC) amiloride sensitive i AT1 cells in-situ, in freshly cut lung slices, and Ussing chamber to record short circuit current from AT1 cells confluents monolayers. We use pClamp, amplifiers, and converters by molecular devices (San Jose, CA, USA) for patch clamp experiments and hardware and software by physiologic instruments (Reno, NV, USA). for imaging and immunostaining experiments we use a Nikon microscope in conjunction with NIS Element for images acquisition and analysis. Lung Wet/Dry ratio, Alveolar fluid clearance, RTP-CR and western blotting data will be presented as aggregate.