Nuclear Patch-Clamp Electrophysiology of Lysosomes

Background of Lysosomal Nuclear Patch-Clamp Electrophysiology

Nuclear patch-clamping is very similar to patch-clamping the cytoplasmic membrane, but this method presents unique challenges and limitations. The development of patch clamp, fluorescence microscopy, and high-resolution electron microscopy has redefined the current concept of ion transport across the nuclear envelope. Researchers interested in signal transduction pathways that affect gene transcription and other processes in the nucleus are increasingly looking at the nuclear envelope from an electrophysiological perspective. The field of nuclear electrophysiology has improved prospects with the fusion of electrophysiology and cell/molecular biology approaches. Because the nuclear membrane provides relatively easy access to the cytoplasmic and luminal sides of the membrane, and allows tight control of ionic and ligand conditions, nuclear patch clamp techniques are currently used to analyze lysosomal ion channels, and this method represents a novel heterologous systems to express endolysosomal channels and transporters.

Fig. 1. Membranes, ion channels, and transporters of the nuclear envelope. (Matzke AJ, et al., 2010)

Lysosomal Nuclear Patch-Clamp Electrophysiology Services

Evidence from animal cells suggests the presence of Ca²⁺ channels and transport proteins in the nuclear membrane. In contrast, lysosomes are intracellular Ca²⁺ storage organelles. Our bioscientists use nuclear patch clamp techniques to identify intact ion channels and transporter proteins in the inner lysosomal membrane and with the outer nuclear membrane. At CD Biosciences, we focus on providing our global customers with efficient lysosomal nuclear patch-clamp electrophysiology services to extensively analyze the ion channels in the lysosomal nuclear membrane and the possible ion gating of the nuclear pores.

We successfully applied this technique to characterize the hTPC2 channel of the lysosome. We developed a customized protocol for the lysosomal patch-clamp technique to characterize the lysosomal hTPC2 channel.

(1) The DT40TKO cell line stably expressing hTPC2 lacks two intracellular Ca²⁺ channels, namely functional InsP3R and RyR.

(2) Using this technique, they detected a ~220 pS single-channel current activated by NAADP with K⁺ as the osmotic ion.

(3) Data analysis and reporting.

Why Choose Us

• Easy access to cytoplasm and luminal side.
• Tightly control ligand conditions.
• High time resolution.
• Simple agreement.
• High signal-to-noise ratio.

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

1. Matzke AJ, et al. (2010) Ion channels at the nucleus: electrophysiology meets the genome. Mol Plant. 3(4):642-52.