Electroneutral transporter monitoring solution is the ONLY method for monitoring directly and in-situ electroneutral and transporter activity. The DHM® optical measurement provides quantitative measurement of the co-transporter activity.
Measurements with DHM® are performed in-situ, that is to say in native cells. There is no need of using any fluorescence, nanoparticle or radioactive tracers, or to add a complex and expensive set-up on top of your microscope. It applies to many experimental protocols and many type of cells.
Monitoring of NKCC1 and KCC2, two neuronal co-transporters involved in chloride homeostasis, has been demonstrated and published [Jourdain, 2011]. DHM is the first imaging technique offering the possibility to directly monitor the activities of the two neuronal co-transporters, NKCC1 and KCC2 in-situ.
The electroneutral transporter monitoring solution combines a transmission DHM®, and a software for acquisition and analysis. It can be completed with an optional motorized XYZ stage and with a fluorescence module enabling to compare DHM® measurements with fluorescence data. Output data are compatible with ImageJ for instance for more extensive and detailed analysis.
Use of DHM® for quantitative monitoring of Electroneutral Transporter enables to investigate new research paths. It provides novel research possibilities to be published in high impact factor scientific journals.
The Electroneutral Transporter Monitoring solution is a simple and direct system for monitoring co-transporters. Electroneutral transporters cannot be measured using electrophysiology. Fluorescence methods as well cannot be applied for such problematic. Use of radioactive tracers is the sole alternative method to DHM®.
Use of radioactive tracers requires complex experimental protocols with load and washout. Heavy radioactive protection measures are necessary.
Moreover, for instance in the co-transporter problematic, radio-tracers are linked to only one of the co-transports, for instance to KCC2 in the KCC2 and NKCC1 case.
One can note that. additionally, DHM enables monitoring of electrogenic transporter while radiotracers cannot.
Feature | DHM® | Radiotracers |
Measure of co-transporter activity | ||
No sample preparation | ||
Simple experimental protocol |
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Electrogenic transporter activity |
Electro-neutral transporte activity of several channels of different type of cells has been investigated, and results have been assessed using pharmacology methods and patch clamp.
NKCC1 and KCC2 are two neuronal co-transporters involved in Cl– homeostasis and cell volume regulation. Dysfunction of these two co-transporters has consequences on neuronal excitability, leading, in extreme cases, to pathogenesis of seizures and chronic neuropathic pain. The fact that NKCC1 and KCC2 co-transporters have electro-neutrally positive and negative charges makes the study by electrophysiology impossible. Study is performed using DHM® on primary cultures of cortical neurons prepared from E17 OF1 mice embryos.