Fluorescence microscope with image analysis system for the cytogenetic studies of germ cells.

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We request a high-performance fluorescence microscope with an image analysis system. This microscope makes it possible to visualize specific molecules and structures within a cell, aided by the binding of fluorescent dyes. It is fully automated and integrated with its own computer system, which processes images for recording and analysis. Its function of overlaying multiple images allows the user to investigate the association between various molecules and structures. This requested equipment will facilitate and extend the following three research programs, which will use mouse models with the aim of understanding how genetic material is transmitted through the germ cells of parents to their babies: (1) A great many female germ cells (named oocytes) are initially produced, but the majority of them are eliminated before birth, thus limiting the reserve of future oocytes. Our first goal is to identify the cause and mechanism of this major oocyte loss. (2) Embryos inherit DNA from both parents. Paternal and maternal DNAs, however, differ with respect to their epigenetic marks. These differences are maintained in somatic cells but erased in primordial germ cells (future sperm or oocytes) in embryos, and new epigenetic marks are established during germ cell differentiation depending on the carrier's sex. Our second goal is to determine the timing of the establishment of epigenetic marks in the germ cells and identify the enzymes involved. (3) Mitochondria are cytoplasmic organelles essential for respiration in all living cells. They carry their own DNAs (mtDNAs) and are exclusively inherited from the mother. mtDNA proliferation, active in oocytes, stops after fertilization and restarts in embryos before their implantation. Our third goal is to define the regulatory mechanism of mtDNA proliferation in oocytes and embryos. Any errors in these three biological processes would result in infertility, embryonic death, or genetic disease. The trainees in our laboratories will get the opportunity to carry out experiments involving image analyses, which will help them to acquire expertise in any field of biology.