A novel 65 kDa RNA-binding protein in squid presynaptic terminals

Author Posting. © The Author(s), 2009. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Neuroscience 166 (2010): 73-83, doi:10.1016/j.neuroscience.2009.12.005.A polyclonal antibody (C4), raised against the head domain of chicken myosin Va, reacted strongly towards a 65 kDa polypeptide (p65) on western blots of extracts from squid optic lobes but did not recognize the heavy chain of squid myosin V. This peptide was not recognized by other myosin Va antibodies, nor by an antibody specific for squid myosin V. In an attempt to identify it, p65 was purified from optic lobes of Loligo plei by cationic exchange and reverse phase chromatography. Several peptide sequences were obtained by mass spectroscopy from p65 cut from SDS-PAGE gels. BLAST analysis and partial matching with ESTs from a Loligo pealei data bank indicated that p65 contains consensus signatures for the hnRNP A/B family of RNA-binding proteins. Centrifugation of post mitochondrial extracts from optic lobes on sucrose gradients after treatment with RNase gave biochemical evidence that p65 associates with cytoplasmic RNP complexes in an RNA-dependent manner. Immunohistochemistry and immunofluorescence studies using the C4 antibody showed partial co-labeling with an antibody against squid synaptotagmin in bands within the outer plexiform layer of the optic lobes and at the presynaptic zone of the stellate ganglion. Also, punctate labeling by the C4 antibody was observed within isolated optic lobe synaptosomes. The data indicate that p65 is a novel RNA-binding protein located to the presynaptic terminal within squid neurons and may have a role in synaptic localization of RNA and its translation or processing.REL, JCR and JEM received financial support from the Fundação de Amparo à Pesquisa do Estado de Sao Paulo (FAPESP), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and the Fundação de Apoio ao Ensino, Pesquisa e Assistência do Hospital das Clínicas da FMRP-USP (FAEPA). JAD received financial support from the RI-INBRE Program Grant #P20RR016457 from the Nation Center for Research Resources, NIH, Bethesda, MD. DTPL, LC, SBFT, EJRV and MMAB were recipients of research fellowships from FAPESP and CNPq. REL and JEM received Productivityin- Research fellowships from CNPq

Randomised controlled trial of transanal endoscopic microsurgery versus endoscopic mucosal resection for large rectal adenomas (TREND Study)

Cellular mechanisms in basic and clinical gastroenterology and hepatolog

New insights into V-ATPase functioning: the role of its accessory subunit Ac45 and a novel brain-specific Ac45 paralog

Contains fulltext : 91340.pdf (publisher's version ) (Open Access)Radboud Universiteit Nijmegen, 19 december 2011Promotor : Martens, G.J.M.194 p

Molecular Probing of the Secretory Pathway in Peptide Hormone-Producing Cells

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Novel insights into V-ATPase functioning: distinct roles for its accessory subunits ATP6AP1/Ac45 and ATP6AP2/(pro) renin receptor

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Bone attenuation on routine chest CT correlates with bone mineral density on DXA in patients with COPD

Chronic obstructive pulmonary disease (COPD), although primarily a disease of the lungs, is associated with extrapulmonary effects such as muscle weakness and osteoporosis. Fractures owing to osteoporosis cause significant morbidity and mortality, particularly in patients with COPD. To prevent osteoporotic fractures, it is important to diagnose osteoporosis in an early stage and to start anti-osteoporotic therapy in at-risk patients. Because routine chest computed tomography (CT) is increasingly used to assess the extent of emphysema and airways disease in patients with COPD, we investigated whether simple attenuation measurement of the thoracic spine on routine chest CT may provide useful information on bone health in patients with COPD. Fifty-eight patients with moderate to very severe COPD were included in our study. The average attenuation of thoracic vertebrae 4, 7, and 10 on chest CT was correlated with the lowest bone mineral density (BMD) of the hip and lumbar spine (L-1 to L-4) on dual-energy X-ray absorptiometry (DXA) in patients with COPD. The inter- and intra-observer variabilities of the attenuation measurements were low as shown by Bland-Altman plots. Pearson's correlation coefficient between the average attenuation of the three thoracic vertebrae and the lowest BMD of the hip and lumbar spine was high (r = 0.827, p <0.001). A receiver-operating characteristic (ROC) analysis of the area under the curve for osteoporosis was 0.969 (p <0.001), corresponding to an attenuation threshold of 147 Hounsfield Units (HU). In conclusion, our data demonstrated that bone attenuation measured on routine chest CT correlated strongly with BMD assessed on DXA in patients with COPD. Routine chest CT may provide useful information on bone health in patients with COPD. (C) 2012 American Society for Bone and Mineral Research

Assessing the utility of Magneto to control neuronal excitability in the somatosensory cortex

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Novel vertebrate- and brain-specific driver of neuronal outgrowth

Contains fulltext : 235666 .pdf (Publisher’s version ) (Open Access