A method for cryopreservation and activation of spermatozoa of ovoviviparous fish Scorpene xushi

An ultra-low temperature preservation and ovoviviparous technology, which is applied in the field of marine organisms and achieves the effects of simple preservation and activation methods, good stability and easy operation.

Active Publication Date: 2021-07-27
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

At present, the research on sperm maturation, activation of in vivo fertilization and sperm preservation in Scorpius xu's is still blank

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Cryopreservation:

[0018] 1) In December 2017, 2 males of flounder flounder in mating period were collected in cages in Yantai, and their venous blood was obtained. The blood menstrual flow will be obtained at 8000r / min, and the supernatant will be centrifuged to obtain serum. At the same time, 0.5 semen during mating period will be obtained ml, stored in a refrigerator at 4°C.

[0019] 2) Take 1 ul of the above-mentioned semen and mix it with 400 ul of the above-mentioned serum for activation. Observe the motility rate of the sperm under a microscope, which is above 95%, and use it for cryopreservation experiments;

[0020] 3) Prepare the diluent, 0.7% NaCl and the above step 1) and mix the obtained serum uniformly at a volume ratio of 1:5 as the diluent, and then add DMSO to the diluent to make the final concentration of DMSO reach 10% (volume ratio) , obtain antifreeze solution and store it in a refrigerator at 4°C;

[0021] 4) Mix the antifreeze solution and seme...

Embodiment 2

[0025] 1) In December 2018, 4 gonad-developed males of Flounderus xushii were purchased in Qingdao market, and their venous blood was obtained. The blood menstrual flow will be obtained at 8000r / min, and the supernatant will be centrifuged to obtain serum. At the same time, 0.8ml of semen during mating period will be obtained. , were stored in 0-4 ℃ refrigerator.

[0026] 2) Take 1 ul of the above-mentioned semen and mix it with 400 ul of the above-mentioned serum for activation. Observe the motility rate of the sperm under a microscope, which is above 95%, and use it for cryopreservation experiments;

[0027] 3) Prepare the diluent, 0.7% NaCl and the above step 1) to obtain the serum in a volume ratio of 1:5 and mix it evenly as the diluent, then add DMSO to the diluent to make the final concentration of DMSO reach 12% (volume ratio) , obtain antifreeze solution and store it in a refrigerator at 4°C;

[0028] 4) Mix the antifreeze solution and semen obtained above at a volum...

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Abstract

The invention belongs to the technical field of marine biology, and in particular relates to a method for ultra-low temperature preservation and activation of sperm of the ovoviviparous fish Scorpionus scorpionus. The sperm obtained from the mature stage of the sperm of the male scorpionfish of Xu's flatfish is evenly mixed with the freezing liquid in the ratio of 1:4-1:5, and then the gradient cooling is carried out for freezing, and then the frozen mixed liquid is moved to liquid nitrogen, that is, the sperm freezing is realized. Ultra-low temperature long-term storage; the gradient cooling freezing is that the mixed solution is pre-cooled at 0-4°C for 2-5min, then frozen to -80-100°C at a speed of 10-12°C / min, and then 15-18°C / min Freeze at ‑150°C. The invention establishes a method for ultra-low temperature preservation of ovoviviparous fish sperm and a sperm activation method, and provides a new method and a new idea for cultivating new varieties of marine fish.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, in particular to a method for cryopreservation and activation of the sperm of ovoviparous fish Scorpius xushii. Background technique [0002] Sebastes schlegelii (Hilgendorf, 1880), belongs to the family Scorpidae of Scorpiomorpha. Sebastes schlegelii is a kind of warm-temperate demersal ovoviviparous fish, and it is an important economically farmed fish species in northern my country. Xu's flat scorpionfish is a typical oviparous fish, fertilized in vivo, embryos develop in vivo to larvae and then produced outside the body. Xu's flat scorpionfish mates every autumn. At this time, the sperm of the male fish is mature, and the next spring is fertilized, and the embryo develops and fertilizes in vivo. Because of the special development, maturation and fertilization methods of the Scorpius spermatozoa, the sperm cryopreservation and activation methods are very different from those of ovipa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
Inventor 刘清华李军王学颖徐世宏
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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