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The Expression of Imprinting Gene Inpp5f-v3 during Mouse Brain Development

Author: HouJing
Tutor: ChenYan
School: Harbin Institute of Technology
Course: Biochemistry and Molecular Biology
Keywords: development of brain Inpp5f-v3 in situ hybridization Northern blotting real-time RT-PCR
CLC: Q75
Type: Master's thesis
Year: 2009
Downloads: 7
Quote: 0
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Abstract


Inpp5f-v3 is a paternally expressed imprinted gene; locate in distal mouse chromosome 7. Inpp5f-v3 is a transcriptional variant of Inpp5f. Inpp5f is an inositol phosphatase gene that is biallelically expressed in the mouse. Inositol phosphatases are involved in endocytosis and exocytosis at nerve terminals of mammal. Inpp5f perform important functions during the mouse development. In order to study the expression and regulation effects of Inpp5f in the development of mouse brain, the primer of Inpp5f-v3 was designed at first, the Inpp5f-v3 fragment was obtained by RT-PCR and subcloned into plasmids pBluescriptⅡK S, then synthesized DIG-UTP-labeled single-stranded RNA probes. Northern blotting analysis reveals a 2.7 kb transcript that is expressed only in brain. We used in situ hybridization and real-time RT-PCR to characterize the expression of Inpp5f-v3 gene during mouse embryonic development. During the mouse embryonic development, Inpp5f-v3 gene was expressed in the olfactory bulb, cerebral cortex, hindbrain and external germinal layer (EGL) of cerebellum. After birth, the expression was enhanced in cerebellum and hippocampus. Strong signal was observed both in the EGL and the internal granule layer (IGL), weak signal was observed in the purkinje cells. In the hippocampus, Inpp5f-v3 gene expressed in the pyramidal cells of the CA1 and CA3 and the granule cells of the dentate gyrus. In addition, Inpp5f-v3 was also expressed in granule cell layer, mitral cell layer and periglomerular cells of the olfactory bulb. These results indicate that its spatial and temporal expression patterns are developmentally regulated, Inpp5f-v3 may play an important role in the development of brain and nervous system, Inpp5f-v3 may involve in the growth, differentiation and maturity of the cerebellar granule cells.

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