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, presents workshops and talks, serves on several NIH committees and helps coordinate lab+office safety for NIDCD (including annual inspections). Projects (RCDC=Neuro, Aud Sys): NIDCD: Bechara Kachar lab: The Core recently has been training and collaborating extensively with the new fellow, Robstein Chidavaenzi, on electron microscope tissue preparation, ultramicrotomy, immunolabeling, and electron microscope operation. We have been working together with Robstein and Dr. Kachar to design individual experiments related to his main current project examining spectrins in stereocilia rootlets. The Core also has been involved in several projects either recently published or in preparation, with Dr. Kachar's lab. In addition, the Core also gives help with other projects of Dr. Kachar, including preparing tissue and cutting sections of the cochlea and of the rhizarian protist, Allogromia. We also assist and work with Evan Krystofiak on some aspects of use of the electron microscope and related instruments. (1G&2B-Per Aud/Vest S&F) Paul Sieving/Yong Zeng/Vijay Camasamudram/Hongman Song/Ron Bush NIDCD/NEI: The Core prepares samples and examines retinal tissue with EM for 2 different studies on retinal diseases, and consults on possible functions, involving Bush/Yong/Vijay and Bush/Hongman, respectively. Matt Kelley lab: Previously we worked with fellow, Helen May-Simera (Matt Kelley lab; now in the University of Mainz in Germany), on the ciliary proteins, BBS8 and IFT20, as part of a study on 'ciliary' proteins and their association with the PCP protein Vangl. The Core performed EM immunogold labeling plus EM on the developing mouse cochlea. One paper was published but a second one is still in preparation. In addition, we advised his lab on protocols for perfusing mice and helped Hanna Sherrill with a perfusion earlier this year. (1F-Dev-Aud Sys) Tom Friedman/Inna Belyantseva lab: The Core currently is training and advising Inna Belyantseva on use of the new electron microscope and related techniques, to optimize ultrastructural studies of hair cells and stereocilia. In addition, we have an earlier collaboration that is part of a manuscript in preparation, with Melanie Barzik, Tom Friedman (NIDCD) and John Hammer NHLBI: we studied the localization of the motor protein, Myo18A in the cerebellum, utilizing EM methods. (1G&2B-Per Aud/Vest S&F) Richard Chadwick/Alexander Cartagena-Rivera: Earlier this year Ron Petralia trained Alex in the various techniques of Scanning Electron Microscopy and discussed strategies of using the methodology in his studies. We also have discussed possible collaborations in the near future. (1F-Dev-Aud Sys) Doris Wu lab: We recently began planning a study with Doris Wu and Tao Jiang to look at changes in EM structure of stereocilia and kinocilia and the cuticular plate in the utricle of mutant mice. We hope to process samples for them soon. (1G&2B-Per Aud/Vest S&F) Catherine Weisz will join the institute in October and we have discussed possible collaborations. 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 Julien Thevenon and Laurence Faivre (Universit de Bourgogne, Dijon, France) and involves a SALM1-associated human pathology (manuscript in press). 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. Currently, we are examining ectosome/exosome involvement in sonic hedgehog secretion and in the effects of sonic hedgehog on mitochondria and other structures in neurons. We also have one sonic hedgehog paper in revision at this time. In addition, we published a recent review paper (Petralia is first author), on invaginating projections in synapses of animals. NICHD Chris McBain/Ken Pelkey/Megan Wyeth lab: This year we published a paper in Neuron on pentraxins and glutamate receptors associated with parvalbumin neurons in the hippocampus. Currently, we continue our studies on glutamate receptors at inhibitory synapses in the hippocampus. NHGRI Ellen Sidransky lab: Currently, we are working on two studies using EM ultrastructure. We are doing a study with Nahid Tayebi on ultrastructural changes in different regions of the brain in mice with combinations of Parkinson's and Gaucher diseases. In addition, we recently began looking at EM changes in neuronal cultures with Elma Aflaki. NINDS Jeff Diamond/Jun Zhang. We are collaborating on a methods paper for improved immunogold localization of glutamate receptors in the retina; this will be a video paper for JoVE. We also advise and assist Jun Zhang on preparation techniques for EM, and may have further collaborations in these studies. NIHMS Song Jiao and Zheng Li: We continue to work on a study with them on autophagy and synaptic vesicles; currently we are working out new ways to determine if synaptic vesicles are incorporated into autophagosomes. NICHD Dax Hoffman/Lin Lin: We now are planning a new study on localization of DPP6, a potassium channel auxiliary protein with EM immunogold; we published an earlier paper with them on this protein (Sun et al., 2011). Outside NIH: We are working with Maria Rubio of the Auditory Research Group at the University of Pittsburgh on deafness effects on auditory synapses in the cochlear nucleus using EM Tomography. Specifically, we are comparing synapses at auditory nerve/bushy cell-endbulb synapses in normal versus ear-plugged animals and animals after earplug removal. We also have discussed doing a collaboration with Rebecca Seal of the University of Pittsburgh and Catherine Weisz (new to NIDCD) on synaptic changes in hair cells in VGLUT3 knockout mice. We recently published a paper with Gavin Rumbaugh/Massimiliano Aceti (Scripps Institute) on SynGAP mutant effects on synapses of somatosensory cortex (associated with schizophrenia), (one of our electron micrographs made the cover photo of the journal issue of Biological Psychiatry). Now we are working on a new study of SynGAP in the cortex. We recently published both a research paper and review paper on NMDA receptor trafficking with Martin Horak of the Czech Republic. We expect to continue some research with his lab in the next fiscal year. We also train and advise Kazuyo Takeda (FDA/CBER/Microscopy and Imaging Core Facility, and NHLBI guest scientist) on various EM methods, and we are discussing a possible collaboration. In addition to collaborations, Ron Petralia currently serves as a consultant on several studies, and some of these might turn into collaborations eventually: Taro Fujikawa (Japan)-glutamate receptor function in hair cell synapses; Hugh Xia (LSU Health Sciences Center)-PP1/I-2 location in neuronal compartments; Wenqin Luo (Dept. Neurosci., Perelman Sch. Med., Univ. Penn.) and Yuan-Xiang Tao (New Jersey Med. Sch., Rutgers)- glutamate receptor changes in the spinal cord dorsal horn; Jeffrey Erickson (LSU Health Sciences Center)-hippocampal synapses.
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