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Transgenic zebrafish with specific myelin sheath removal capacity as well as preparation method and application thereof

A zebrafish and transgenic technology is applied in the field of zebrafish model preparation to achieve the effect of improving transgenic efficiency

Inactive Publication Date: 2014-08-06
GENERAL HOSPITAL OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the animal models currently used for the study of demyelination, myelination injury and / or remyelination mechanism and related drug screening still need to be improved

Method used

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  • Transgenic zebrafish with specific myelin sheath removal capacity as well as preparation method and application thereof
  • Transgenic zebrafish with specific myelin sheath removal capacity as well as preparation method and application thereof
  • Transgenic zebrafish with specific myelin sheath removal capacity as well as preparation method and application thereof

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Embodiment 1

[0074] 1. Experimental materials and methods

[0075] 1. Experimental animals

[0076] Wild-type zebrafish were cultured according to a standardized protocol (see: Westerfield, M. The zebrafish book: a guide for the laboratory use of zebrafish (Brachydanio rerio) 1993, Eugene, OR: M. Westerfield, which is hereby incorporated by reference in its entirety) (AB strain), water temperature 28.5°C, light / dark cycle 14h / 10h, adult zebrafish embryos were collected after spawning, cultured in E3 hatching solution, measured by hours of fertilization (hours post-fertilization, hpf) or days of fertilization (days post-fertilization, dpf) indicates the developmental stage of embryos and juveniles.

[0077] 2. Preparation of transgenic zebrafish

[0078] Transgenic zebrafish Tg(mbp:nfsB::EGFP) was prepared by the Tol2 system.

[0079] 2.1 Construction of mbp-nfsB-EGFP recombinant plasmid

[0080] The promoter of the zebrafish myelin basic protein (mbp) gene was amplified by PCR. The len...

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Abstract

The invention discloses transgenic zebrafish with specific myelin sheath removal capacity as well as a preparation method and an application thereof. The preparation method comprises the following steps: recombinant plasmid mbp-nfsB-EGFP and a Tol 2 transposase mRNA (messenger ribonucleic acid) are jointly introduced into wild zebrafish; and a zebrafish strain with stable inheritance is cultured and obtained, and the zebrafish strain with stable inheritance is the transgenic zebrafish with specific myelin sheath removal capacity, wherein the nucleotide sequence of an mbp gene promoter carried in the recombinant plasmid mbp-nfsB-EGFP is represented as SEQ ID NO.1. With adoption of the method, the transgenic zebrafish which can specifically remove myelin sheath, cause myelinoclasis and has stable inheritance capacity can be prepared and obtained effectively, under the action of metronidazole, oligodendrocyte of juvenile transgenic zebrafish can be removed specifically and the juvenile transgenic zebrafish has pathologic change of myelinoclasis together with decreased motor function, therefore, the transgenic zebrafish can provide a forceful animal model for a regulatory mechanism for viviperception of myelin sheath damage and regeneration and related drug screening.

Description

technical field [0001] The invention relates to a preparation method and application of a zebrafish model, in particular to a transgenic zebrafish capable of specifically removing myelin sheath tissue, and a preparation method and application thereof. More specifically, the present invention relates to a method for preparing a transgenic zebrafish that can specifically remove myelin tissue, a transgenic zebrafish that can specifically remove myelin tissue, and research on the mechanism of demyelination, myelin damage and / or regeneration thereof , and use in the screening of drugs for the treatment of demyelination, myelination injury, and / or for remyelination. Background technique [0002] Myelin sheath is a membrane structure that protects, supports and nourishes axons. In the central nervous system, myelin sheath is formed by mature oligodendrocytes protruding out to surround axons; in the peripheral nervous system, it is composed of Schwann cells . The main function of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N5/10A01K67/027A61K49/00
Inventor 魏世辉李玉皓方杨武
Owner GENERAL HOSPITAL OF PLA
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