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Packaged high-flux measurement method for expression change of gene and corresponding protein in hippocampus during nicotine addiction, withdrawal and addiction reconstruction stage

A protein expression, hippocampal technology, applied in the field of life science research

Active Publication Date: 2020-10-27
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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  • Abstract
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Problems solved by technology

However, there is currently no suitable group sample mixing method that can meet the requirements of high throughput and small sample volume to complete this experiment

Method used

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  • Packaged high-flux measurement method for expression change of gene and corresponding protein in hippocampus during nicotine addiction, withdrawal and addiction reconstruction stage
  • Packaged high-flux measurement method for expression change of gene and corresponding protein in hippocampus during nicotine addiction, withdrawal and addiction reconstruction stage
  • Packaged high-flux measurement method for expression change of gene and corresponding protein in hippocampus during nicotine addiction, withdrawal and addiction reconstruction stage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] 1. Instruments and reagents

[0115] Nanodrop2000 ultra-micro spectrophotometer (Thermo Scientific, USA), Roche LC96 fluorescent quantitative PCR instrument (Roche, Switzerland), multi-functional microplate reader (Molecular Devices, USA), Bio-Rad protein electrophoretic transfer system (Bio-Rad, USA) , Fusion Fx7 multifunctional gel imaging system (French VILBERLOMRMAT company).

[0116] Nicotine (TRC, purity 99.5%), normal saline (domestic), fluorescent quantitative reagent FastStart EssentialDNA Green Master (Roche, 6402712001), RNAiso Plus, DNA polymerase, PCR primers etc. (Japan TAKARA company), agar powder (Spain Amresco Packing). Nicotinic acid (TRC, purity 99.5%), normal saline (domestic), Anti-Nicotinic Acetylcholine Receptor alpha 3 antibody (UK abcam, ab103956); Anti-Nicotinic Acetylcholine Receptor alpha 4 antibody (UK abcam, ab124832); Anti-Nicotinic Acetylcholine Receptor alpha 5 antibody (abcam, UK, ab157470); Anti-Nicotinic Acetylcholine Receptor alpha...

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Abstract

The invention discloses a packaged high-flux measurement method for the expression change of eight genes and corresponding nAChRs protein in hippocampus during a rat nicotine addiction, withdrawal andaddiction reconstruction stage. The method comprises the following steps of: on the basis of a CPP (conditioned place preference) device, establishing a rat nicotine addiction, withdrawal and addiction reconstruction model; during a period and finally, obtaining a nicotine addiction, withdrawal and addiction reconstruction rat and a corresponding control rat; and killing the rat, peeling the hippocampus, preparing cDNA and total protein, and detecting the expression changes of eight CHRNs genes and eight nAChRs protein. According to the method, three stages, including smoking addiction, withdrawal and smoking again, can be simulated, the expression change of eight CHRNs genes and corresponding nAChRs protein are detected so as to be favorable for researching signal mechanisms under different addiction states, and the method has an important function for researching a nicotine addiction related molecular mechanism and developing a nicotine withdrawal treatment measure.

Description

technical field [0001] The invention belongs to the field of life science research, and specifically relates to a rat model based on simulating different stages of smoking addiction, stages and relapse, and detecting the expression changes of 8 kinds of CHRNs genes and corresponding 8 kinds of nAChRs proteins in hippocampus tissue Panelized high-throughput assay method. Background technique [0002] Smoking and health is an important research topic in the field of international public health. Nicotine is the main addictive substance in cigarette smoke. It is very important to study the mechanism of nicotine addiction and discover the biomarkers of nicotine addiction for smoking and health issues. [0003] In the process of addiction research, it is first necessary to construct a reliable animal addiction model, and then detect the expression changes of nicotine addiction and withdrawal-related genes based on the constructed animal model, and then explore the mechanism and p...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6851G01N33/68A01K67/02
CPCC12Q1/6883C12Q1/6851G01N33/6893A01K67/02C12Q2600/158G01N2800/307G01N2333/70571A01K2267/03A01K2227/105A01K2207/20C12Q2531/113C12Q2565/125C12Q2545/101C12Q2563/107
Inventor 王红娟陈欢付亚宁韩书磊刘彤侯宏卫胡清源
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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