| 研究実績の概要 |
The purpose of this research is to understand the mechanism of mammalian sperm hyperactivation that is required for sperm to successfully fertilize oocytes. Hyperactivation is characterized by increased sperm motility that is caused by calcium influx and the cAMP pathway. I will study two testis-specific genes implicated in the cAMP pathway, Tcp11 and Tcp11l3, in mice. Tcp11 was previously reported to form a complex in mouse sperm with Protein Kinase A, a key mediator of cAMP signaling. My research demonstrated that Tcp11 is not present in mature mouse sperm from the epididymis, but is present in germ cells of the testis. Deletion of Tcp11 causes a subfertile phenotype due to slow moving sperm, which is consistent with Tcp11 functioning in the cAMP pathway. In contrast, deletion of Tcp11l3 leads to no discernable effect on male fertility, which suggest Tcp11 may compensate for Tcp11l3 function and not vice versa. Some of this work was published in Biology of Reproduction (Castaneda et al 2020, PMID: 31837139).
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| 今後の研究の推進方策 |
I have successfully analyzed Tcp11 knockout males and generated Tcp11l3 knockout males and have determined that Tcp11l3 is not required for spermatogenesis. Tcp11 KO males are subfertile, and I suspect that Tcp11l3 might slightly compensate for the absence of Tcp11. My future plan is to generate the Tcp11/Tcp11l3 double knockout and examine fertility of these males and characterize spermatogenesis in these mice. My future plan also includes examining the proteome of TCP11 and TCP11L3 with immunoprecipitation (IP) and mass spectrometry (MS). TCP11 and TCP11L3 are proteins with unknown functions. The proteome of these two proteins will allow me to hypothesize about their molecular function. Specifically, IP and MS results will allow me to conclude whether these two proteins have a direct role in the cAMP pathway. I have successfully generated an antibody to TCP11 (Figure 4), that I can use for IP from mouse testis.
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