PURPOSE. RP is a retinal degeneration disorder that is caused by mutations of various genes, including semaphorin-4A (SEM-A4A). A number of retinal diseases, including RP, are associated with light exposure, oxidative stress, and endoplasmic reticulum (ER) stress. In this study, we investigated whether mutant SEMA4A causes retinal dysfunction via light exposure, oxidative stress, and ER stress.METHODS. Mutant SEMA4A (D345H or F350C) was overexpressed in a human retinal epithelium cell line ARPE19. Intracellular localization of mutant SEMA4A was investigated using confocal laser scanning microscopy. The ARPE-19 cells were also irradiated with white light, and expression of 78 kDa glucose-regulated protein (GRP78), a marker of ER stress, and phagocytosis were measured. The cells were treated with an ER stress inducer, tunicamycin, or an oxidative stressor, H 2 O 2 , and cell death was measured. Human SEMA4A mutants were expressed in zebrafish embryos with tunicamycin and mRNA of DNA damage-inducible transcript 3 (ddit3) was measured as an ER stress marker.RESULTS. Mutant SEMA4A was localized in the ER, whereas wild type (WT) SEMA4A was observed in cell membranes. The expression of GRP78 was increased by mutant SEMA4A following light irradiation, and phagocytosis was suppressed in mutant SEMA4A-transfected cells. Mutant SEMA4A induced susceptibility to ER stress and oxidative stress. In zebrafish, human mutant SEMA4A increased ddit3 mRNA compared with WT under the ER stress condition.
CONCLUSIONS.Our results suggest that mutations in SEMA4A may cause susceptibility to light exposure, oxidative stress, and ER stress, which may be involved in the progression and pathology of RP. (Invest Ophthalmol Vis Sci. 2012;53:6729-6737)
A notable advantage of zebrafish as a model organism is the ease of gene knockdown using morpholino antisense oligonucleotide (MO). However, zebrafish morphants injected with MO for a target protein often show heterogeneous phenotypes, despite controlling the injection volume of the MO solution in all embryos. We developed a method for estimating the quantity of MO injected into each living morphant, based on the co-injection of a control MO labeled with the fluorophore lissamine. By applying this method for knockdown of cardiac troponin T (tnnt2a) in zebrafish, we could efficiently select the partial tnnt2a-depleted zebrafish with a decreased heart rate and impairment of cardiac contraction. To investigate cardiac impairment of the tnnt2a morphant, we performed fluorescent cardiac imaging using Bodipy-ceramide. Cardiac image analysis showed moderate reduction of tnnt2a impaired diastolic distensibility and decreased contraction and relaxation velocities. To the best of our knowledge, this is the first report to analyze the role of tnnt2a in cardiac function in tnnt2a-depleted living animals. Our combinatorial approach can be applied for analyzing the molecular function of any protein associated with human cardiac diseases.Electronic supplementary materialThe online version of this article (doi:10.1007/s12033-013-9664-6) contains supplementary material, which is available to authorized users.
Hemangioblastomas of the cerebellopontine angle (CPA) that emerge extra-axially from the peripheral nervous system are extremely rare. We report a case of hemangioblastoma of the CPA evaluated by pseudocontinuous arterial spin labeling (pCASL). The high rate of tumor blood flow determined using pCASL provided additional useful information for the differential diagnosis of the CPA tumors in this patient.
Objective
This study evaluated the efficacy of 3-dimensional fluid-attenuated inversion recovery (3D FLAIR) for detecting intradural ecchordosis physaliphora (EP).
Methods
We retrospectively determined the presence or absence of intradural EP on 3D FLAIR for 3888 consecutive patients, classifying the EP as “classical” or “possible” and analyzing the prevalence, size, and presence or absence of an intraosseous stalk. Where available, magnetic resonance cisternography images were compared with the 3D FLAIR images.
Results
Intradural EP was identified in 50 patients (1.3%): 36 (0.9%) classical and 14 (0.4%) possible. The classical EPs were significantly larger than the possible EPs (P < 0.01). Nine EPs (18.0%) showed an osseous stalk. Magnetic resonance cisternography was performed for 19 EPs (16 classical, 3 possible), detecting all 16 classical EPs but none of the possible EPs.
Conclusions
Classical EPs were detected by 3D FLAIR as with magnetic resonance cisternography. The 3D FLAIR findings suggested a new type of possible EP variant previously unreported.
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