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Atohla expression diagram method in zebra fish lateral line hair cell regeneration process

A regeneration process, hair cell technology, applied in biochemical equipment and methods, microbial determination/inspection, instruments, etc., can solve problems such as lack of atoh1 and inability to form hair cells

Pending Publication Date: 2018-10-12
HANGZHOU NORMAL UNIVERSITY
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Problems solved by technology

Previous studies have shown that atoh1 is a hair cell fate determinant, overexpression of atoh1 can induce excess hair cells, and loss of atoh1 hair cells cannot form

Method used

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  • Atohla expression diagram method in zebra fish lateral line hair cell regeneration process
  • Atohla expression diagram method in zebra fish lateral line hair cell regeneration process
  • Atohla expression diagram method in zebra fish lateral line hair cell regeneration process

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Embodiment Construction

[0020] A method for the expression pattern of atoh1a in the regeneration process of zebrafish lateral line hair cells. The method uses atoh1a antisense nucleotide probes to observe cisplatin-induced zebrafish lateral line hair cells by whole embryo in situ hybridization The expression pattern of atoh1a during death and regeneration, and the spatial relationship between atoh1a expressing cell populations and mature hair cells in the center of the neuromast were observed by in situ hybridization and antibody staining and co-staining, and further observed by adding exogenous Notch signaling inhibitors Changes in the number of regenerating hair cells in the presence of increased expression of Atoh1a.

[0021] The synthesis of the atoh1a antisense nucleotide probe is specifically to design primers to amplify the atoh1a mRNA fragment, connect the Ez-T vector, and construct a plasmid. After the plasmid is linearized, it is purified and recovered. Digoxigenin-labeled antisense probes ...

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Abstract

The invention discloses an atohla expression diagram method in a zebra fish lateral line hair cell regeneration process. The method utilizes an atohla antisense nucleotide probe, by using a whole-mount in situ hybridization method, the atohla expression diagram is observed in the process that the fish lateral line hair cells die and regenerate under the induction of cis-platinum, the spatial relationship between expression cell groups of the atohla and nerve-hillock center mature hair cells is observed by using the method of co-dyeing of hybridization in situ and antibody staining, a Notch signal inhibitor is added through an outer source to further observe the change of regenerated hair cell quantity under the circumstance that atohla expression is increased, and in vivo experimental evidence is provided to determine that cell groups marked by the atohla are precursor cells in the zebra fish lateral line hair cell regeneration process.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a method for expressing patterns of atoh1a in the regeneration process of zebrafish lateral line hair cells. Background technique [0002] The neurogenic gene atoh1a, which belongs to the bHLH family of transcription factors, plays a key role in the fate determination of vertebrate hair cells. Previous studies have shown that atoh1 is a hair cell fate determinant, overexpression of atoh1 can induce excess hair cells, and loss of atoh1 hair cells cannot form. During the formation of the lateral line system in zebrafish, FGF signaling can initiate the expression of Notch signaling pathway ligand delta A. Both Delta A and Delta D activate the Notch signaling pathway in adjacent cells and inhibit the expression of atoh1a, and the cells expressing atoh1a will eventually develop into hair cells (hair cells), and the adjacent Notch signal-activated cells will develop into helper ce...

Claims

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Application Information

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IPC IPC(8): C12Q1/6841C12Q1/6888G01N33/569
CPCG01N33/56966C12Q1/6841C12Q1/6888C12Q2600/158
Inventor 芈肖肖
Owner HANGZHOU NORMAL UNIVERSITY
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