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P2‐049: MELATONIN TREATMENT ATTENUATES GLUCOSE DYSREGULATION AND GLIOSIS IN HYPERGLYCEMIC NON‐TG MICE
Author(s) -
Robinson Alice,
Asuni Ayodeji A.,
Salman Rania
Publication year - 2014
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2014.05.722
Subject(s) - endocrinology , medicine , carbohydrate metabolism , melatonin , gliosis , glucose transporter , neurodegeneration , neuroinflammation , insulin resistance , circadian clock , insulin , glut3 , circadian rhythm , biology , glut1 , inflammation , neuroscience , disease
frontotemporal dementia. It is still unclear how this mutation causes disease, however altered expression of C9ORF72 transcripts are present in C9ORF72 patients suggesting that loss of function is a potential pathogenic mechanism of disease. C9ORF72 encodes a protein of unknown function. In this study we have examined the expression and localization of C9ORF72 over a developmental timecourse both in vivo and in vitro in order to provide some insight into the normal role of the protein. Methods: Primary neuron and glia cultures were derived from embryonic and postnatal rodents, grown for up to 21 days and fixed in PFA or harvested for Western blotting over a timecourse of 1, 3,7 14 and 21 days. Brains were harvest from mice at E15, E18, P1, P7, P14 and P28 and processed for Western blotting or fixed in PFA. Immunolabelling and Western blotting were performed with antibodies against C9ORF72 in addition to cytoskeletal proteins, axonal and dendritic markers and cellular organelles. Results: C9ORF72 was consistently expressed in both neurons and glia from early in development. C9ORF72 was predominantly absent from the nucleus but had a punctate expression in the cytoplasm and throughout the neuropil in vivo. In vitro the protein was localized to discrete vesicles (approximate diameter 0.6mm), which extended into both axons and dendrites. C9ORF72 immunoreactive-vesicles extended beyond the microtubule cytoskeleton into actin rich structures, such as filapodia. C9ORF72 was not localized to Golgi apparatus or mitochondria but shared some co-localization with components of the endosome/lysosome system. C9ORF72 immunoreactivity rarely localized to synaptic puncta in vitro or in vivo. Conclusions: These data suggest that C9ORF72 is a vesicular protein that is expressed throughout development and into adulthood. Protein expression is present in both neuron and glial populations. C9ORF72 immunoreactive vesicles are found throughout the cell including in actin rich structures and may be involved in membrane trafficking. Determining the normal role of C9ORF72 protein may help to determine the role it plays in disease.