精子SSP411蛋白在受精卵首次卵裂中的功能机制研究

本团队在国际上首次发现并证明ssp411是精子特异表达基因，其缺失表达导致精子异常及受精卵首次卵裂延迟。首次卵裂是胚胎发育的起始，需精卵协作，但精子的贡献甚少被研究，蛋白含量低不易被检测是原因之一。我们用基因编辑技术构建了HA-tag 转基因鼠，解决了微量蛋白检测难题，并用该技术重构了基因敲除鼠模型，解决了原敲除鼠的技术缺陷。在以上工具鼠研究基础上，我们提出“SSP411是精子特异的MTOC组分，在受精卵首次卵裂中发挥重要作用”。我们将通过HA-tag 转基因小鼠，鉴定SSP411在配子及受精卵不同期中的表达及与MTOC共定位，证明SSP411是来自精子的受精卵MTOC组分；通过基因敲除模型，证明精子SSP411蛋白缺失致MTOC功能缺陷是首次卵裂延迟的机制。以上研究，我们将解析精子提供SSP411蛋白是受精卵MTOC执行首次卵裂功能所必须，为首次卵裂的精子贡献提供有力的实验证据。

The first cleavage is the beginning of embryonic development and requires the cooperation of sperm and egg, but the contribution of sperm is rarely studied. As testis and sperm specifically expression protein, SSP411 was first reported by our team in the world on 2004. Lack the expression of SSP411, causes sperm abnormalities and fertilized eggs first cleavage delay. That means this protein is important in sperm and embryonic development. We construct the HA-tag transgenic mouse with CRISPR/CAS9 to trace protein detection, and construct the gene knockout mouse model with the same technology defects of the original knockout mouse. Based on the above mouse models, we propose that SSP411 is a component of epididymis sperm-specific cytoskeletal protein in MTOC, which plays an important role in the first cleavage of fertilized eggs. We are planning to express SSP411 protein and co-localization with MTOC in gametes and embryos of each stage using the HA-tag transgenic mouse model, to prove that SSP411 is a component of MTOC of fertilized eggs derived from sperm. Then, we are going to use SSP411 gene knockout model to prove one of the mechanism of the first cleavage delay is lack of SSP411 protein. Through the above research, we are going to analyze the SSP411 protein function for proving: sperm is necessary for MTOC to perform the cleavage function in fertilized egg, and sperm proteins contribute to the first cleavage in fertilized egg.