Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction

Katsunori Yanai; Shohei Kaneko; Hiroki Ishii; Akinori Aomatsu; Kiyonori Ito; Keiji Hirai; Susumu Ookawara; Kenichi Ishibashi; Yoshiyuki Morishita

doi:10.3791/61383

Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ure...

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Genetics

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JoVE Journal Genetics

Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction

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07:36 min

August 27, 2020

DOI: 10.3791/61383-v

Katsunori Yanai¹, Shohei Kaneko¹, Hiroki Ishii¹, Akinori Aomatsu^1,2, Kiyonori Ito¹, Keiji Hirai¹, Susumu Ookawara¹, Kenichi Ishibashi³, Yoshiyuki Morishita¹

¹Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center,Jichi Medical University, ²Division of Intensive Care Unit, First Department of Integrated Medicine, Saitama Medical Center,Jichi Medical University, ³Department of Medical Physiology,Meiji Pharmaceutical University

Overview

This article describes a method for evaluating microRNA expression in the kidneys of mice with unilateral ureteral obstruction (UUO) using quantitative reverse-transcription polymerase chain reaction. The technique allows for high accuracy and sensitivity in profiling microRNA expression in various pathological conditions.

Key Study Components

Area of Science

Neuroscience
Biology
Renal Pathology

Background

MicroRNAs play a crucial role in gene regulation.
Unilateral ureteral obstruction (UUO) is a model for studying kidney disease.
Understanding microRNA profiles can provide insights into renal fibrosis.
Current methods may lack sensitivity and accuracy.

Purpose of Study

To develop a reliable method for assessing microRNA expression in renal tissues.
To facilitate research on kidney diseases and their underlying mechanisms.
To improve the understanding of microRNA roles in pathological conditions.

Methods Used

Quantitative reverse-transcription polymerase chain reaction (qRT-PCR).
Sham surgery to prepare the mouse model.
Use of surgical tools for tissue manipulation.
Application of PBS for tissue handling.

Main Results

The method allows for accurate microRNA profiling.
High sensitivity in detecting microRNA levels in kidney tissues.
Facilitates the study of various renal pathologies.
Reduces technical errors compared to traditional methods.

Conclusions

This method provides a valuable tool for kidney research.
It enhances the understanding of microRNA functions in renal diseases.
Future studies can leverage this technique for broader applications.

Frequently Asked Questions

What is the significance of microRNA in kidney research?

MicroRNA regulates gene expression and is crucial for understanding kidney disease mechanisms.

How does this method improve upon existing techniques?

It offers higher accuracy and sensitivity while minimizing technical errors.

What are the applications of this microRNA profiling method?

It can be used in various studies related to renal pathologies and microRNA functions.

What is unilateral ureteral obstruction (UUO)?

UUO is a model used to study kidney disease and its effects on renal function.

What tools are required for the sham surgery?

Surgical scissors, tweezers, and cotton swabs are needed for the procedure.

Can this method be applied to other pathological conditions?

Yes, it is suitable for studying microRNA expression in various kidney diseases.

We describe a method for evaluating the microRNA expression in the kidneys of mice with unilateral ureteral obstruction (UUO) by quantitative reverse-transcription polymerase chain reaction. This protocol is suitable for studying kidney microRNA expression profiles in mice with UUO and in the context of other pathological conditions.

This method makes it possible to provide microRNA expression in kidneys of mice with a wide range of pathological conditions, such as renal fibrosis. The main advantage of this technique is that it enables microRNA expression profiling with high accuracy and sensitivity by using a simple process that saves time and prevents technical error. To perform the sham surgery, use surgical scissors and tweezers to make an incision in the skin at the abdomen and cut the muscle and peritoneal membrane from the bladder to the left lower edge of the ribs.

Moisten two cotton swabs with PBS and carefully pull the intestines to the side. Place the moistened swabs to identify the left kidney and ureter. Close the peritoneal membrane and then the incision with 4-0 nylon.

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