In vitro analysis of mammalian meiosis has remained a significant challenge for many years. These difficulties have arisen due to the interactions that occur between germ cells and the supporting Sertoli cells. Through these interactions, Sertoli cells regulate spermatogenesis: spermatogonial stem cell renewal, entry of spermatogonia into meiosis, and meiotic progression. Due to the importance of meiotic processes in mammalian development, it is important to develop efficient methods for in vitro analyses. A visual marker is needed to monitor the progression of meiosis in vitro. H1t is a testis-specific linker histone selectively expressed in mid to late pachytene spermatocytes (meiotic germ cells) and round spermatids. The goals my project included characterizing these transgenic mice by confirming the expression of the H1t-GFP fusion protein in testis. Secondly, I worked to identify at what point during the meiotic process this transgene is expressed and determine its exact onset within these mice. Additionally, I used okadaic acid (OA), a phosphatase inhibitor, to induce meiotic progression towards metaphase as a method of increasing H1t-GFP expression.
Throughout my experience, I learned how to apply my knowledge of the scientific method into the current field of medicine in an effort to further the understanding of mammalian meiotic processes. I also learned a lot about the breeding and development of transgenic mice. I believe that the experience I have gained from this research will translate into my educational studies, as the problem-solving skills I have gained throughout this project have the potential to be applicable to my future academic career
In vitro analysis of mammalian meiosis has remained a significant challenge for many years. These difficulties have arisen due to the interactions that occur between germ cells and the supporting Sertoli cells. Through these interactions, Sertoli cells regulate spermatogenesis: spermatogonial stem cell renewal, entry of spermatogonia into meiosis, and meiotic progression. Due to the importance of meiotic processes in mammalian development, it is important to develop efficient methods for in vitro analyses. A visual marker is needed to monitor the progression of meiosis in vitro. H1t is a testis-specific linker histone selectively expressed in mid to late pachytene spermatocytes (meiotic germ cells) and round spermatids. The goals my project included characterizing these transgenic mice by confirming the expression of the H1t-GFP fusion protein in testis. Secondly, I worked to identify at what point during the meiotic process this transgene is expressed and determine its exact onset within these mice. Additionally, I used okadaic acid (OA), a phosphatase inhibitor, to induce meiotic progression towards metaphase as a method of increasing H1t-GFP expression.
Throughout my experience, I learned how to apply my knowledge of the scientific method into the current field of medicine in an effort to further the understanding of mammalian meiotic processes. I also learned a lot about the breeding and development of transgenic mice. I believe that the experience I have gained from this research will translate into my educational studies, as the problem-solving skills I have gained throughout this project have the potential to be applicable to my future academic career