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Biology
L'utilizzo di biosensori enzimatica per quantificare endogena ATP o H 2 O 2...
L'utilizzo di biosensori enzimatica per quantificare endogena ATP o H 2 O 2...
JoVE Journal
Biology
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JoVE Journal Biology
Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

L'utilizzo di biosensori enzimatica per quantificare endogena ATP o H 2 O 2 Nel rene

Full Text
12,323 Views
10:00 min
October 12, 2015

DOI: 10.3791/53059-v

Oleg Palygin1, Vladislav Levchenko1, Louise C. Evans1, Gregory Blass1, Allen W. Cowley Jr.1, Alexander Staruschenko1

1Department of Physiology,Medical College of Wisconsin

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article discusses the use of enzymatic microelectrode biosensors for real-time measurement of extracellular cell signaling. The protocols outlined extend the application of these biosensors to detect ATP and hydrogen peroxide in kidney tissues.

Key Study Components

Area of Science

  • Neuroscience
  • Biochemistry
  • Bioengineering

Background

  • Extracellular signaling is crucial for cellular communication.
  • Real-time measurement techniques enhance understanding of cellular processes.
  • ATP and hydrogen peroxide are important signaling molecules in the kidney.
  • Existing methods may not provide the necessary sensitivity or specificity.

Purpose of Study

  • To measure endogenous concentrations of ATP and hydrogen peroxide.
  • To develop protocols for ex vivo and in vivo applications.
  • To improve the understanding of kidney function and signaling.

Methods Used

  • Rehydration and calibration of sensors.
  • Surgical installation of sensors in the kidney.
  • Flushing the kidney through the aorta to remove blood.
  • Perfusion of isolated kidneys with saline for measurements.

Main Results

  • Successful installation of sensors for real-time measurements.
  • Detection of ATP and hydrogen peroxide in kidney tissues.
  • Demonstrated feasibility of ex vivo and in vivo applications.
  • Provided insights into the dynamics of cell signaling in the kidney.

Conclusions

  • Enzymatic microelectrode biosensors are effective for real-time measurements.
  • Protocols can be adapted for various experimental conditions.
  • Further research may enhance understanding of kidney signaling mechanisms.

Frequently Asked Questions

What are enzymatic microelectrode biosensors?
They are devices used to measure specific biochemical substances in real-time.
How are ATP and hydrogen peroxide measured?
Through the installation of sensors in the kidney that detect these molecules.
What is the significance of measuring ATP in the kidney?
ATP plays a key role in cellular energy transfer and signaling.
What does ex vivo mean?
Ex vivo refers to experiments conducted on tissues outside of the living organism.
Can these methods be applied to other organs?
Yes, the protocols may be adapted for use in other biological tissues.
What are the potential applications of this research?
Understanding kidney function, drug effects, and disease mechanisms.
Is this technique suitable for clinical use?
Further validation is needed before clinical applications can be considered.

I biosensori a microelettrodi enzimatici consentono misurazioni in tempo reale della segnalazione cellulare extracellulare in concentrazioni biologicamente rilevanti. I seguenti protocolli estendono le applicazioni dei biosensori alla rilevazione ex vivo e in vivo di ATP e H2O2 nel rene.

L'obiettivo generale di queste procedure è misurare in tempo reale le concentrazioni interstiziali endogene di un TP e del perossido di idrogeno nei reni interi. Ciò si ottiene reidratando e calibrando in primo luogo i sensori per applicazioni ex vivo e in vivo a concentrazioni note di un TP e perossido di idrogeno. Per l'installazione del sensore ex vivo, il rene è stato strumentato chirurgicamente e pulito dal sangue.

Questa procedura viene eseguita lavando il rene attraverso l'aorta. Quindi il rene di ratto viene isolato chirurgicamente, posto in una camera di registrazione e perfuso con soluzione fisiologica. Un TP e il perossido di idrogeno vengono misurati utilizzando sensori ex vivo per la procedura in vivo.

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