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Preparation and use method of improved injection mixture

A technology of injecting mixed and solution, applied in the field of injection, can solve the problems of insignificant effect, promotion of follicle neogenesis, low degree of zebrafish testicular differentiation, etc., and achieve the effect of increasing the effect of medication and reducing waste

Inactive Publication Date: 2019-05-31
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Each injection of exogenous gene is single, and can only regulate the differentiation of zebrafish testis in a certain direction
[0008] When a single exogenous gene is injected, the degree of testicular differentiation in zebrafish is low, and the effect is not significant
[0009] There is no good way to promote follicle regeneration

Method used

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  • Preparation and use method of improved injection mixture
  • Preparation and use method of improved injection mixture
  • Preparation and use method of improved injection mixture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0061] 1), Zebrafish preparation

[0062] (1) Dilute MS222 (anesthetic) to 1x with filtered water.

[0063] (2) Prepare 5-month-old and 18-month-old zebrafish, and use 1x MS222 solution to anesthetize the zebrafish.

[0064] (3) After the zebrafish is incapacitated, fix the zebrafish under a microscope using a grooved foam board.

[0065] 2) In vitro gonad localization of zebrafish:

[0066] The body surface projection position of the injection site should be about 1mm above the intersection of the horizontal line at the upper end of the pectoral fin and the vertical line at the front of the pelvic fin (red dotted line). figure 2 shown.

[0067] 3), injection:

[0068] The injection solution is prepared according to the configuration method provided in the embodiment of the present invention, and the mixed solution of different proportions and the existing Sox9a / b exogenous gene injection solution.

[0069] (1) Mixed solution 1: Prepare mixed solution 1 according to the ...

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Abstract

The invention belongs to the technical field of injection, and discloses a preparation and use method of an improved injection mixture. The improved injection mixture is composed of a transfected liposome vector, a foreign plasmid and an exogenous miRNAs solution in proportion. The preparation method comprises diluting the constructed foreign plasmid using the sterilized ddH2O to the concentrationof 500 ng / [mu]L; diluting the prepared miRNAs to the concentration of 20 pmol / [mu]L using the sterilized ddH2O; then preparing the transfected liposome vector; preparing 2-4[mu]L of mixed solution according to the proportion that the transfected liposome vector: Sox9a exogenous plasmid: exogenous miRNA (one or mixture of more) = 2:1:1 ratio. The use method includes adopting in vitro microinjection of gonadal glands combined with electrotransfection. The differentiation of the testis and the ovary of 6-month zebrafish and the gonad regeneration of the adult fish can be controlled by adding different proportions and different kinds of exogenous miRNAs to the original mixed solution.

Description

technical field [0001] The invention belongs to the technical field of injections, in particular to an improved method for preparing and using an injection mixture. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] In the past decade, zebrafish has become a model of high research value as a carrier for various experiments. Zebrafish is closer to mammals in evolution, and has homology with some mammals such as humans and mice. Gene. [0004] Zebrafish exhibit a certain degree of plasticity in the process of sex differentiation, and at the same time have a strong regulatory mechanism. miRNAs and FSH / cAMP / MAPK / Sox9 signaling pathway can regulate the development and sex differentiation of vertebrate gonads, but the specific regulatory network is not clear. Through a zebrafish in vitro projection injection technique, exogenous injection of genes can affect the zebrafish Regulation of gonadal differentiati...

Claims

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Application Information

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IPC IPC(8): A61K9/08A61K9/00A61K48/00A61K31/7105A61P15/08
Inventor 严继舟杜新路郭会萍王成辉
Owner SHANGHAI OCEAN UNIV
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