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Biology
酵素バイオセンサの使用は、内因性ATPまたはHを定量します 2 O 2腎臓で
酵素バイオセンサの使用は、内因性ATPまたはHを定量します 2 O 2腎臓で
JoVE Journal
Biology
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JoVE Journal Biology
Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

酵素バイオセンサの使用は、内因性ATPまたはHを定量します 2 O 2腎臓で

Full Text
12,326 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.

酵素微小電極バイオセンサーは、生物学的に関連性のある濃度での細胞外細胞シグナル伝達のリアルタイム測定を可能にします。 以下のプロトコルは、バイオセンサーのアプリケーションを、腎臓におけるATPおよびH2O2のex vivoおよびin vivo検出に拡張します。

これらの手順の全体的な目標は、腎臓全体のTPと過酸化水素の内因性間質濃度をリアルタイムで測定することです。これは、まず、ex vivoおよびin vivoアプリケーション用のセンサーを既知のTPおよび過酸化水素濃度に再水和および校正することによって達成されます。ex vivoセンサーの設置では、腎臓を外科的に器具化し、血液から洗浄しました。

この手順は、大動脈を通して腎臓を洗い流すことによって行われます。次に、ラットの腎臓を外科的に分離し、記録チャンバーに入れ、生理食塩水で灌流します。TPと過酸化水素は、in vivo手順のためにex vivoセンサーを使用して測定されます。

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