Thumdee, Patama: The prenatal expression of mRNA and protein of the prion protein gene, PRNP, in sheep. - Bonn, 2007. - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn.
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author = {{Patama Thumdee}},
title = {The prenatal expression of mRNA and protein of the prion protein gene, PRNP, in sheep},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2007,
note = {The expression of the prion protein gene both on mRNA and protein levels were investigated in ovine female reproductive organs and in various tissues of their foetuses during the prenatal stage. Reproductive organs such as ovary, oviduct, endometrium, myometrium and caruncle were collected at the 1st, 3rd and 5th month of pregnancy. Foetal tissues were the whole foetuses at 1 month of age, brain, cotyledon, heart, intestine, kidney, liver, lung and muscle of 2-month-old foetuses. At 3 and 5 months of age the spinal cord and spleen were added. Sheep were categorized as resistant (R1) or high susceptible (R5) to scrapie according to their PRNP genotype. In both genotype groups, the gene transcript was detectable in all stages and all tissues examined by RT-PCR. The gene expression profiles of R1 and R5 groups were similar. Comparisons between reproductive organs demonstrated the highest expression level in caruncle tissue of ewes, whereas the level was high in brain and low in liver of their foetuses. In addition, real-time RT-PCR was performed in immature oocytes, mature oocytes, in vivo embryos at morula stage and in 1-month-old foetuses. The results showed that the relative expression levels of PRNP mRNA in mature oocytes and morula-stage embryos were significantly lower than those in immature oocytes and 1-month-old foetuses (p≤0.05). Fluorescent in situ hybridisation in adult ovaries and 1-month-old foetuses demonstrated the presence of the gene transcript in oocytes, granulosa cells, theca cells, ovarian cortex, ovarian medulla and corpus lutuem of the ovaries, and in brain, vertebral column, dermatome, heart, liver and kidney of the foetuses of both groups. Western blot analyses revealed the immunoreactive bands corresponding to PrPC in all female reproductive tissues as well as their foetuses collected at the 1st month gestation. The PrPC was also detected in all tissues of 2-month-old foetuses. In addition, immunohistochemical staining implicated localisation of PrPC in brain, heart and kidney of 1-month-old foetuses. The PrPC was also found in ovarian cortex and ovarian medulla of the two groups however, it was undetectable in oocytes, granulosa cells, theca cells and corpus luteum, in this study.},
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