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Application of phosphodiesterase 5 activity inhibitor as ripening agent for inducing maturation of zebrafish oocyte

An activity inhibitor, phosphodiesterase technology, applied in the field of phosphodiesterase 5 activity inhibitor and inducing zebrafish oocyte maturation agent, which can solve the problems of poor cell quality and low oocyte fertilization rate.

Pending Publication Date: 2019-10-08
NORTHWEST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some laboratories have achieved success in in vitro maturation-in vitro fertilization-in vitro culture, the oocytes matured in vitro have problems such as low fertilization rate and poor cell quality, so by further research on the oocyte maturation mechanism Research, the in vitro maturation system will be more perfect

Method used

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  • Application of phosphodiesterase 5 activity inhibitor as ripening agent for inducing maturation of zebrafish oocyte
  • Application of phosphodiesterase 5 activity inhibitor as ripening agent for inducing maturation of zebrafish oocyte
  • Application of phosphodiesterase 5 activity inhibitor as ripening agent for inducing maturation of zebrafish oocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the application of Sildenafil Citrate in inducing zebrafish oocyte maturation

[0025] After the zebrafish were anesthetized to death, 10-15 female zebrafish were dissected, and the ovaries were taken out and placed in a petri dish containing 60% by mass Leibovitz L-15 medium diluted with sterile water, and artificially separated and grown completely. Ovum (FG stage; approximately 0.65 mm in diameter). The isolation process usually lasts 3–5 h at room temperature, incubated at 28°C, and treated with drugs. The concentration of the drug Sildenafil Citrate was 10uM, and the treatment time was 14 hours. Compared with the control group, the induced oocyte maturation rate increased by 1.83 times. The maturation rate of oocytes in the control group was about 40%, and that of the Sildenafil Citrate-treated group was about 73%.

Embodiment 2

[0026] Embodiment 2, the application of Tadalafil in inducing zebrafish oocyte maturation

[0027] The concentration of the drug Tadalafil was 10uM, and the treatment time was 14 hours. Others are the same as in Example 1. (Compared with the control group, the maturation rate of induced oocytes increased by 2.3 times. The maturation rate of the control group was about 33%, and the oocyte maturation rate of the Sildenafil Citrate treatment group was about 75%.

Embodiment 3

[0028] Example 3, Application of Avanafil in Inducing Zebrafish Oocyte Maturation

[0029] The concentration of the drug Avanafil was 1 mM, and the treatment time was 14 hours. Others are the same as in Example 1. (Compared with the control group, the induced oocyte maturation rate increased by 1.5 times. The maturation rate of the control group was about 52%, and the oocyte maturation rate of the Sildenafil Citrate treatment group was about 78.5%.

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Abstract

The present invention provides a novel use of a phosphodiesterase 5 activity inhibitor, namely, as a ripening agent for inducing maturation of zebrafish oocyte, which belongs to the technical field ofaquatic products. The phosphodiesterase 5 activity inhibitor comprises Sildenafil Citrate, Tadalafil and Avanafil. The experiment shows that Sildenafil Citrate, Tadalafil and Avanafil exhibit a dose-dependent, time-dependent and period-dependent manner in promoting oocyte maturation, and moreover, the efficiency of inducing oocyte maturation in zebrafish is higher. In addition, the phosphodiesterase 5 activity inhibitor can be used not only for inducing maturation of zebrafish oocyte, but also for inducing maturation of other fish oocytes, so that the phosphodiesterase 5 activity inhibitor has good application prospect.

Description

technical field [0001] The invention relates to a new application of a phosphodiesterase 5 activity inhibitor—that is, as an agent for inducing zebrafish oocyte maturation, and belongs to the technical field of aquatic products. Background technique [0002] Phosphodiesterases (PDEs) have the function of hydrolyzing intracellular second messengers (cAMP, cyclic adenosine monophosphate or cGMP, cyclic guanosine monophosphate), and degrade intracellular cAMP or cGMP, thereby terminating the biochemical effects transmitted by these second messengers . Among them, phosphodiesterase 5 is a phosphodiesterase that specifically hydrolyzes cGMP, and its inhibitors include Sildenafil, Vardenafil, Tadalafil and Avanafil, and are mainly used for the treatment of sexual dysfunction. In recent years, it has been found in mammals that PDE5 is involved in the regulation of mammalian oocyte maturation, but no similar reports have been found in fish. This patent mainly discloses the applica...

Claims

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Application Information

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IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/01
Inventor 李建真白琳
Owner NORTHWEST NORMAL UNIVERSITY
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