Ronald S. Petralia and Ya-Xian Wang have run the Advanced Imaging Core facility of NIDCD since October 2011. This core is dedicated primarily to training and collaborating with other scientists in NIDCD in studies utilizing electron microscopy (EM). Also, to a limited extent, some work is done with other NIH institutes or outside NIH. In addition, Ronald Petralia regularly reviews papers for professional journals and reviews many various documents for NIDCD PIs, reviews grants, writes review papers and letters of recommendation, serves on several NIH committees and helps coordinate lab+office safety for NIDCD (including the Clean Sweep this year). This year, we experienced a significant interruption (for about 2 months) due to the move from building 50 to 35A. In addition, we switched to a new, advanced transmission electron microscope that includes EM Tomography. Installation and set-up of this EM took considerable time. And this has entailed extensive, ongoing training on the new microscope system and methods. Projects (RCDC=Neuro, Aud Sys): NIDCD: Matt Kelley: We worked with fellow, Helen May-Simera (now in NEI), on the ciliary proteins, BBS8 and IFT20, as part of a study on 'ciliary'proteins and their association with the PCP protein Vangl2;The Core has been performing EM immunogold labeling plus EM on the developing mouse cochlea. For this study, the Core also has been involved in the extensive cochlea tissue preparation, prepared previously in collaboration with Kate Szarama. A manuscript received favorable reviews and is currently in revision for the journal Development. We also are performing studies on another ciliary protein. Finally, we recently started planning a study with Weise Chang. (1F-Dev-Aud Sys) Bechara Kachar: The Core recently has been training the new fellow, Robstein Chidavaenzi, on electron microscope tissue preparation, ultramicrotomy, immunolabeling, and electron microscope operation. The Core also has been involved in several projects either recently published or in preparation, with Dr. Kachars lab. The first was with Taro Fujikawa on the kainate receptors of the outer hair cells and their afferent terminals;it involved previous training and research. The findings were published in Hearing Research this year. The second project was with Janaina Brusco, on EM ultrastructure of synapses in the posterodorsal medial amygdala. This paper was published in the Journal of Comparative Neurology this year. The third project is with Seham Ebrahim. In late 2013, we performed tissue preparation, ultramicrotomy, immunolabeling, staining, and EM of auditory and vestibular tissue, especially looking at the structure of normal stereocilia and comparing this to the structure of stereocilia in espin mutant mice. The Core also has been working with Dr. Kachar on studies involving the hair cell synaptic ribbon, especially of a mutant mouse for Vglut3, and we hope to develop this further with the use of EM Tomography. The Core also gives help with other projects of the lab. (1G&2B-Per Aud/Vest S&F) Tom Friedman/Inna Belyantseva: The Core currently is training Inna Belyantseva on use of the new electron microscope and related techniques. (1G&2B-Per Aud/Vest S&F) Stephan Brenowitz: The Cores previous extensive work with postdoctoral fellows from Dr. Brenowitzs laboratory, Shan He and Milos Sedlacek (including mentoring of a summer student) was terminated when Dr. Brenowitz left NIH in late 2013, to start a new position at Janelia. However, for the study with Dr. He, on ion channels of the cartwheel cell synapses from the dorsal cochlear nucleus, we did publish a paper in The Journal of Neuroscience this year. (1A-Cen Aud S&F) Lisa Cunningham/Lindsey May/Andrew Breglio: The Core has been working extensively now on 2 separate projects for this laboratory. We have been performing morphological analyses of utricle exosomes with EM for a study with Lindsey May. As a part of this study, the Core helped to mentor a summer student, Tyra Martinez;she presented a summer poster of her findings in August. Also, recently, we performed EM Tomography on EM material from this study. We also have been preparing samples of cochlea for Serial Block Face Imaging using a scanning electron microscope with a built-in Ultramicrotome. (2B-Vest S&F) Katie Kindt: The Core has been working on structural changes in ribbon synapses from Zebrafish hair cells. Recently, this has included study of these synapses using EM Tomography. Yong Zeng/Vijay Camasamudram/Hongman Song/Ron Bush NIDCD/NEI: The Core has prepared samples and examined retinal tissue with EM for 2 different studies on retinal diseases, involving Yong/Vijay and Hongman, respectively. The Core currently has worked with Gail Seabold (formerly NIDCD/currently in the Graduate Partnerships Program/OITE), on synaptic adhesion-like molecule 1 (SALM1) trafficking in neurons in collaboration with a lab in France and involves a SALM1-associated human pathology (manuscript in preparation). Other NIH institutes: NIA /Mark Mattson/Pamela Yao lab: Projects include: endocytic accessory proteins in neurons;another looks at sonic hedgehog in synapse development and function. We will present a poster at the Society for Neuroscience National Meeting on this in November. In addition, we published 2 review papers (Petralia is first author on both), on aging synapses and the evolution of aging mechanisms in animals. The Core continues to be involved in research studies with this group. NICHD Chris McBain/Ken Pelkey/Megan Wyeth lab: The Core worked on a project looking at the role of a kainate receptor auxiliary protein in hippocampal circuitry. This involved preparation of tissue (KO and WT mice), immunogold labeling, and EM, as well as training of the fellow, Megan. Earlier this year, we published a paper on this in The Journal of Neuroscience. Currently, we are working on another research project related to this topic. NHGRI Ellen Sidransky/Nahid Tayebi: Currently, we are working on an ultrastructural study of changes in different regions of the brain in mice with combinations of Parkinsons and Gauchers diseases. NHLBI John Hammer lab: Last year, we studied the localization of the motor protein, myosin My18A in the cerebellum, utilizing EM methods. It is not clear when this study will be completed. NIHMS: Last year we worked on a study with Song Jiao and Zheng Li, on autophagy and synaptic vesicles affected by BAD-BAX-caspase;we expect that this study will be completed and published in the near future. Outside NIH: We published a paper with Paul Worley, Jay Baraban and Irving Reti of Johns Hopkins University, on Narp in nociception in press in the Journal of Neuroimmunology. Others include: with Gavin Rumbaugh (Scripps Institute) on SynGAP mutant effects on synapses of somatosensory cortex (associated with schizophrenia), in press in Biological Psychiatry;one with Jaroslav Blahos of the Institute of Molecular Genetics in the Czech Republic examining associations between 2 variants of the metabotropic glutamate receptor, mGluR1, in press in Neuropharmacology;and one with Houhui Xia from LSU on the neuronal phosphoproteome. Finally, we also have a paper with Nathalie Sans of the University of Bordeaux in France in the final stage of acceptance for Cell Reports, concerning the role of Scribble1 in NMDA receptor recycling. In addition, we recently submitted an invited review paper on NMDA receptor trafficking with Nathalie Sans and Martin Horak (Czech Republic), to Frontiers in Neuroscience. Finally, we are working on a new project with Martin on NMDA receptor trafficking.
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