Expression and Localization of Keap1 During Amelogenesis in the Developing Molar Germ of Rats

서지정보

ㆍ발행기관 : 대한구강생물학회 ㆍ수록지정보 : International Journal of Oral Biology / 43권 / 4호
ㆍ저자명 : Sun-Hun Kim, Yong-Ouk You, Hyun-Mi Ko, Hyun-Jin Kim

목차

Introduction
Materials and Methods
Animals
Preparation of tissue sections and morphologicalanalysis
Isolation of RNA from the maxillary molar tooth germs
RT-PCR
real-time PCR
Western blot analysis
Immunofluorescent staining
Results
Histological findings
Keap1 mRNA expression
Keap1 Western blot
Immunofluorescent findings
Discussion
References

영어 초록

The objective of this study was to examine the expression pattern of Kelch-like ECH-associated protein 1 (Keap1) in the maxillary 2nd molar germs of rats. We used the maxillary 2nd molar germs in rats’ pup at postnatal day 3 (bell stage), 6 (crown formation stage) and 9 (root formation stage). The investigation on mRNA and protein levels were done using reverse transcription - polymerase chain reaction and western blot. Localization of Keap 1 in the maxillary 2nd molar germs were revealed through immunofluorescence staining. Keap1 from the maxillary 2nd molar germs were mostly manifested on postnatal day 3 and dramatically decreased on postnatal day 6 and 9 at mRNA and protein levels, while amelogenin and ameloblastin increased during the development of maxillary 2nd molar germs. During immunofluorescence analysis, the strong immunoreactivity against Keap1 was detected in the apical side of ameloblasts at the presecretory and secretory stages. However, Keap1 expression was hardly observed in the ameloblasts at the maturation stage. These results shows that Keap1 is strongly expressed in the presecretory and secretory ameloblasts of amelogenesis, and suggest that Keap1 may be a crucial molecule for the regulatory mechanisms tasked with the formation of enamel layer.

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