Background of Lysosomal Channels Study
The cell membranes of intracellular organelles make up the majority of the entire cell membrane and contain a large number of transmembrane proteins, including many ion channels. Among these intracellular channels, those located in the endolysosomal membrane are essential controls for many physiological functions, including endolysosomal ion and pH homeostasis, regulation of endolysosomal resting membrane potential, catabolic export of amino acids, vesicle transport and vesicle fusion. In addition, dysfunction or loss of function of these ion channels is closely associated with the pathophysiology of human diseases. The patch-clamp technique is the standard method for analyzing plasma membrane ion channels, but due to the small size and intracellular localization of lysosomes, this method is not suitable for electrophysiological studies of intracellular lysosomes. To overcome this problem, scientists have successfully developed a unique lysosomal patch-clamping technique that can be used to study lysosomal channels in their native environment by directly recording isolated lysosomes.
Fig. 1. Schematic showing ion channels that have been measured with the endolysosomal patch-clamp technique. (Zhang X, et al., 2020)
Lysosomal Channel Study Service Based on Lysosome Patch-Clamp Technique
Typical lysosomes are less than 0.5 nm in diameter and it is difficult to observe and record them under a light microscope. Our biological scientists use pharmacological or genetic methods to expand endolysosomes to a repairable size, and obtain intact and expanded endolysosomes by mechanical separation or differential centrifugation. CD BioSciences has established a proven lysosomal patch-clamping technology platform to directly record isolated lysosomes to study lysosomal channels in their native environment.
Our technology platform allows efficient endolysosomal patch-clamping of ion channels from heterologous expression as well as from native cells such as cardiomyocytes, hippocampal neurons or macrophages. Here, we provide a comprehensive scheme to analyze lysosomal ion channels via the endolysosomal patch-clamping technique.
(1) Expression of endolysosomal channels in heterologous systems.
(2) Preparation of primary cells.
(3) Enlargement of endolysosomal vesicles.
(4) Design and production of high-end glass electrodes.
(5) Isolation and release of intracellular lysosomal vesicles.
(6) Acquisition and analysis of diaphragm clamp data.
Why Choose Us
- Allows direct measurement of endolysosomal ion channels in the native environment.
- Allows efficient and advanced recording of endolysosomal ion channels.
- A visually controlled direct membrane clamp technique.
- A team with basic knowledge and extensive experience in membrane clamp methods.
- Experimental cycle time of 1 week and detailed data analysis.
- No biochemical separation or purification of endolysosomal vesicles is required.
- Suitable for exploring and characterizing already known endolysosomal ion channels.
- No specific equipment is required.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
Reference
- Chen CC, et al. (2017) Patch-clamp technique to characterize ion channels in enlarged individual endolysosomes. Nat Protoc. 12(8):1639-1658.