Ivestigation Of Fertile Ability Of Sperm Derived From HSL Knock Out Mice Testis By ICSI

Hormone sensitive lipase plays an important role in fat metabolism,not only has the decomposition activity of triacylglycerol,but also has the activity of lipase of diacylglycerol, acylgiycerol and cholesterol ester.According the analyzing of the physiological function of HSL deficient male mice,we found that HSL also plays an important role during the process of spermatogenesis.HSL deficient male mice have few sperm,biopsy showed that there is almost no sperm in their epididymis but some cell debris.However,there arc a few of sperms in their testis.The key course of infertility of HSL deficient male mice is the scare number of sperm. However,whether the knock out of HSL affects the fertilization ability of sperms is not known, and whether the few sperms in the testis could fertilize the normal eggs is not reported in literatures yet.In order to test the fertilization ability of sperm derived from the testis of HSL deficient mice,Intracytoplamic sperm injection(ICSI) technology that is inject the sperm into the cytoplasmic of oocyte was used in this experiment and the development of zygotes as a testing standard fertilization ability of sperm.After ICSI,if the zygotes developed into 2-cell stage embryos,it will demonstrate that the sperm of HSL deficient mice have the ability of fertilization,the deficient of HSL does not affects the function of sperm and the infertility of HSL deficient mice just because of the scare number of sperm.On the other hand,if the zygotes are not fertilized,it will suggest that the deficiency of HSL makes the sperm dysfunctional.A total of five HSL-/- male mice were sacrificed,each experiment sacrifice one mouse to obtain sperm in order to do ICSI and the normal oocytes are derived from KM female mice.A total of 73 oocytes are injected in five injection cycle and 8 of them are developed into 2-cell stage embryos.One of the 8 embryos is good in morphology,but rests are cleavage unequally and the blastomeres are in different size.The result of the experiment reveals that the sperm of HSL-/- deficient male mice are functional which means it has the ability of fertilization and the zygotes could develop into 2-cell stage embryo in vitro.However,whether the embryos have full developmental potential need to carry out follow-up experiment,such as embryo transfer can be confirmed.