38 research outputs found
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
Systems genetics identifies a role for Cacna2d1 regulation in elevated intraocular pressure and glaucoma susceptibility
Glaucoma is a multi-factorial blinding disease in which genetic factors play an important role. Elevated intraocular pressure is a highly heritable risk factor for primary open angle glaucoma and currently the only target for glaucoma therapy. Our study helps to better understand underlying genetic and molecular mechanisms that regulate intraocular pressure, and identifies a new candidate gene, Cacna2d1, that modulates intraocular pressure and a promising therapeutic, pregabalin, which binds to CACNA2D1 protein and lowers intraocular pressure significantly. Because our study utilizes a genetically diverse population of mice with kno
Cross-ancestry genome-wide association analysis of corneal thickness strengthens link between complex and Mendelian eye diseases
Central corneal thickness (CCT) is a highly heritable trait associated with complex eye diseases such as keratoconus and glaucoma. We perform a genome-wide association meta-analysis of CCT and identify 19 novel regions. In addition to adding support for known connective tissue-related pathways, pathway analyses uncover previously unreported gene sets. Remarkably, >20% of the CCT-loci are near or within Mendelian disorder genes. These included FBN1, ADAMTS2 and TGFB2 which associate with connective tissue disorders (Marfan, Ehlers-Danlos and Loeys-Dietz syndromes), and the LUM-DCN-KERA gene complex involved in myopia, corneal dystrophies and cornea plana. Using index CCT-increasing variants, we find a significant inverse correlation in effect sizes between CCT and keratoconus (r =-0.62, P = 5.30 × 10-5) but not between CCT and primary open-angle glaucoma (r =-0.17, P = 0.2). Our findings provide evidence for shared genetic influences between CCT and keratoconus, and implicate candidate genes acting in collagen and extracellular matrix regulation