| 研究実績の概要 |
I investigated the morphological changes of both the uterus and the extraembryonic tissues through analysis of 3D rendered images of post-implantation embryos. Proof of principle was determined by making 3D reconstructions of heart tissues from serially sectioned 9.5 dpc embryos, scanned at high resolution, aligned by unique and precise automated software, and then delineated from surrounding tissues by manual tracing throughout 241 serial sections using the free open-source software FIJI/TrakEM2. Gene expression analysis with reverse-transcription quantitative PCR of iron regulatory genes expression was performed, and analysis through immunofluorescence of non-pregnant and pregnant stages of 4.5-9.5 days. SLC40a1 protein was upregulated in non-pregnant females is particularly high in the LE and GE of the uterus. Female SLC40a1 transgenic uteri were collected at several stages of the uterine cycle and stained for LacZ. This showed that SLC40a1 is most highly expressed during the estrus cycle. High expression of Lacz staining is exhibited in the luminal epithelium and glandular epithelium with little or no expression in the underlying stromal tissue. Concurrently, analysis of the ovary and oviduct revealed consistent expression of SLC40a1-driven LacZ independent of estrus cycle, suggesting a differential expression mechanism of ovarian SlC40a1 vs uterine SLC40a1. DAB staining of both non-pregnant and pregnant female WT mice exhibited differential expression patterns consistent with promoter driven Lacz staining and immunofluorescence.
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