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Biomarker, detection method thereof and application of biomarker in epilepsy pathology research and anti-epilepsy drug screening

A biomarker and epilepsy technology, applied in the biological field, can solve the problems of low epilepsy incidence and difficult observation, and achieve the effect of efficient judgment

Pending Publication Date: 2021-10-22
浙江赛微思生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the deficiency that the zebrafish epilepsy model with lower incidence and frequency of epilepsy in the prior art is not easy to observe the occurrence of epilepsy, and provides a biomarker and its detection method, which can efficiently judge the pathology of epilepsy This method has the advantages of fast, objective and accurate

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  • Biomarker, detection method thereof and application of biomarker in epilepsy pathology research and anti-epilepsy drug screening

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] A biomarker, the biomarker is related to the pathological characteristics of zebrafish epilepsy, and its detection method comprises the following steps:

[0071] Step 1: Fix 5-7-day-old zebrafish juveniles with their backs facing upwards and place them in the sample chamber of the stage of the confocal imaging system with low-melting point agarose. Use a 5X objective lens to perform dynamic imaging of GCaMP fluorescence. The data acquisition frequency is not less than 20 Frame / second, continuous recording for 3 to 5 minutes.

[0072] Step 2: Import the acquired image data into the image processing software ImageJ, select the optic tectum (optictectum) brain region (such as figure 1 The area marked by the white dotted line) is used as the analysis object, and then the data of the target area is imported into MATLAB for analysis, and the data of the fluorescence intensity of all pixels in the area changing with time are extracted.

[0073] Step 3: Set the activation stat...

Embodiment 2

[0085] Evaluate the efficacy of antiepileptic drugs by using neural excitation clusters as epilepsy biomarkers.

[0086] Drug treatment: treatment with 500 μM n-heptanol solution (heptanol; CAS number: 111-70-6).

[0087] The juvenile zebrafish stxbp1b gene mutant was fixed in the sample chamber of the stage of the confocal imaging system with low-melting point agarose, and the imaging recording was carried out according to the method in Example 1.

[0088] After the recording is completed, add the drug solution to be evaluated on the surface of the sample, wait for 30 minutes, wait for the drug to penetrate into the juvenile fish, and perform imaging recording again according to the above method.

[0089] Export the data recorded twice and analyze as above.

[0090] Such as Figure 4 It was shown that after the epileptic zebrafish model was treated with antiepileptic drugs, the neural excitation clusters were significantly inhibited, and the size and number were restored to...

Embodiment 3

[0093] Evaluate the efficacy of antiepileptic drugs by using neural excitation clusters as epilepsy biomarkers.

[0094] Drug treatment: treatment with 10 μM propofol solution (propofol; CAS number: 2078-54-8).

[0095] The juvenile zebrafish stxbp1b gene mutant was fixed in the sample chamber of the stage of the confocal imaging system with low-melting point agarose, and the imaging recording was carried out according to the method in Example 1.

[0096] After the recording is completed, add the drug solution to be evaluated on the surface of the sample, wait for 30 minutes, wait for the drug to penetrate into the juvenile fish, and perform imaging recording again according to the above method.

[0097] Export the data recorded twice and analyze as above.

[0098] Such as Image 6 It was shown that after the epileptic zebrafish model was treated with antiepileptic drugs, the neural excitation clusters were significantly inhibited, and the size and number were restored to no...

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Abstract

The invention provides a biomarker, a detection method thereof and application of the biomarker in epilepsy pathology research and anti-epilepsy drug screening, the biomarker is related to zebra fish epilepsy pathology characteristics, and the biomarker is a nerve excitation cluster; the nerve excitation cluster is a set of adjacent nerve excitation clusters in time and space obtained when MATLAB analysis is performed on image data obtained by performing GCaMP fluorescent dynamic imaging on zebra fish; and the nerve excitation cluster is a set of at least three connected excitation state pixel points in a single-frame image. According to the invention, a GCaMP dynamic imaging technology and a zebra fish hereditary epilepsy model are combined, data are subjected to visualization and quantitative analysis by adopting an image data processing pipeline based on MATLAB, a local nerve excitation cluster is used as an epilepsy biomarker, efficient judgment of the epilepsy pathology condition of a nervous system is realized, and the biomarker has the advantages of rapidness, objectivity, accuracy and the like and can be used for screening high-throughput antiepileptic drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a biomarker, its detection method and its application in epilepsy pathology research and antiepileptic drug screening. Background technique [0002] At present, the epilepsy model based on zebrafish is widely used in the research of epilepsy pathological mechanism and the screening of antiepileptic drugs. [0003] Zebrafish-based epilepsy models usually use movement trajectories or electrophysiological signals as epilepsy biomarkers to judge epilepsy and evaluate the efficacy of antiepileptic drugs, but such methods are only applicable to models that exhibit stable high-frequency epilepsy , such as the chemically induced epilepsy model and the Dravet syndrome model with scn1Lab mutation, the incidence of epilepsy is about 80%, and the frequency of epilepsy is about 1 time / minute. [0004] However, other zebrafish genetic epilepsy models exhibit lower seizure incidence an...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6456
Inventor 刘静梁萌萌余伟师
Owner 浙江赛微思生物科技有限公司
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