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Transgenic zebrafish capable of specifically clearing myelin sheath tissue and its preparation method and application

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

Inactive Publication Date: 2016-08-17
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 capable of specifically clearing myelin sheath tissue and its preparation method and application
  • Transgenic zebrafish capable of specifically clearing myelin sheath tissue and its preparation method and application
  • Transgenic zebrafish capable of specifically clearing myelin sheath tissue and its preparation method and application

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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) (Strain AB), the water temperature was 28.5°C, and the light / dark cycle was 14h / 10h. The embryos were collected from adult zebrafish after spawning, and cultured in E3 incubation solution. Hours post-fertilization (hpf) or fertilization days (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) were prepared by the Tol2 system.

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

[0080] The zebrafish myelin basic protein (mbp) gene promoter was amplified by PCR method, one promo...

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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 that can specifically remove myelin 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 clear myelin tissue, a transgenic zebrafish that can specifically clear myelin tissue, and its mechanism in demyelination, myelin damage and / or regeneration. , and for use in screening drugs for the treatment of demyelination, myelin damage and / or for remyelination. Background technique [0002] Myelin sheath is a membranous structure that protects, supports and nourishes axons. In the central nervous system, myelin sheath is formed by mature oligodendrocytes extending out of protrusions to surround axons; in the peripheral nervous system, it is composed of Schwann cells . The main function of myelin is to insulate an...

Claims

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Application Information

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