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). NIDCD: Doris Wu lab: We have been working extensively on studies with Doris Wu, Sharda Yadav, and Kazuya Ono, to look at changes in TEM and SEM structure of sterocilia and kinocilia and the cuticular plate in the utricle of mutant mice. With Sharda Yadav, we have been working on TEM of OMX2 knockout mouse, examining changes in the kinocilium basal body in the utricle and cochlea; this material has been difficult to obtain and study because these mice die at or shortly before birth. We also are working with Sharda on an OMX2 overexpressing mouse, examining changes in patterns of stereocilia and kinocilia in the utricle and cochlea with SEM. With Kazuya Ono, we have been concentrating on using SEM to study changes in the pattern of stereocilia and kinocilia of an OMX2 mutant mouse. Lisa Cunningham lab: Recently, we have returned to a TEM project that we started 2 years ago, looking at effects of heat shock on the release/uptake of exosomes in the utricle. We are doing these studies in collaboration with the postBac IRTA fellow, Nora Welsh, and Andrew Breglio from the lab. We also are working with Shimon Francis on an SEM preparation that will be used for a cover photo. Katie Kindt lab: The Core continues periodically to use TEM to study the hair cells and associated supporting cells of the zebrafish neuromasts and utricle. Recently we have been performing a detailed search for gap junctions in these structures. Bechara Kachar lab: The Core continues to collaborate extensively with the fellow, Robstein Chidavaenzi; We have been examining spectrins and associated proteins in stereocilia rootlets. We work specifically together on various TEM studies involving immunogold labeling and alternative fixation protocols; we also discuss regularly the LM data overall. The Core also has been involved in several projects either recently published or in preparation, with the Kachar lab. In the late Fall, 2015, the Core assisted Jun Zhao to obtain TEM images for his project. In the Spring, 2016, we provided extensive assistance to Willy Sun and Evan Krystofiak, on ultramicrotomy and identification of hair cell structures, for their TEM. Paul Sieving/Yong Zeng/Vijay Camasamudram/Hongman Song/Ron Bush NIDCD/NEI: The Core prepares samples and examines retinal tissue with EM for different studies on retinal diseases, and consults on possible functions, involving Bush/Yong/Vijay and Bush/Hongman, respectively. Earlier this year, we published a paper. Matt Kelley lab: In previous years, we worked with fellow, Helen May-Simera (now in U. Mainz, Ger.) on ciliary proteins. The Core performed TEM immunogold labeling on developing mouse cochlea. 1 paper published; 2nd is in review. Tom Friedman/Inna Belyantseva lab: The Core continues to train/advise Inna Belyantseva on TEM techniques, to optimize ultrastructural studies of hair cells and stereocilia. We assist Inna with tissue preparation, e.g., freeze substitution. Also, the Core is involved in a collaboration with Inna on TEM tomography. The Core worked with Gail Seabold (formerly NIDCD/currently in OITE), on SALM1 trafficking in neurons in collaboration with Thevenon & Faivre (U. Bourgogne, France) and involves a SALM1-associated human pathology (published last Fall). More recently, we have been working with Gail to complete work that she did when she was in NEI on retinal SALMs. Other NIH institutes: NIA /Mark Mattson/Pamela Yao lab: We published a paper on sonic hedgehog and axon growth (end of 2015; in collaboration with Jennifer Lippincott-Schwartz). We recently published studies on ectosomes/exosomes in sonic hedgehog secretion, and we have a paper in review on effects of sonic hedgehog on mitochondria (with Jennifer Lippincott-Schwartz). In addition, we published a comprehensive review on diversity of spine synapses of animals. NINDS Katherine Roche/Richa Lomash lab: We recently began a pilot study using TEM to examine distribution of neurolastin. We are looking at structural changes in the neuron structure in the brain of the mutant. NICHD Chris McBain/Ken Pelkey/Megan Wyeth lab: Last year we published a paper in Neuron on pentraxins and glutamate receptors. Although Megan moved to California, we continue to collaborate on studies of inhibitory neurons in the hippocampus. -- Jennifer Lippincott-Schwartz/Carolyn Ott: collaborations with Mark Mattson/ Pamela Yao of NIA. Jennifer/ Carolyn recently moved to HHMI/Janelia. NHGRI Ellen Sidransky lab: Currently, we are working on studies using EM ultrastructure, mainly with Nahid Tayebi, related to Parkinsons and Gauchers diseases. We also did TEM in neuronal cultures with Elma Aflaki. NINDS Jeff Diamond/Jun Zhang. We published a method for improved immunogold localization of glutamate receptors in retina (JoVE video). We also advise/assist Jun Zhang on preparation techniques for EM, and expect further collaborations. NIHMS Song Jiao and Zheng Li: We worked with them on synaptic vesicles incorporation into autophagosomes (and may continue). NICHD Dax Hoffman/Lin Lin: We did extensive testing of immunogold labeling for localization of DPP6, a potassium channel auxiliary protein with EM immunogold. Outside NIH: Rumbaugh/Massimiliano Aceti (Scripps Institute): We recently submitted a paper on function of Syngap1 for review, based on work that we did with them earlier this year. Last year, we published a paper with them on SynGAP effects on synapses (associated with schizophrenia) (incl. cover photo of Biol. Psych.). We recently began a study with Jaroslav Blahos (Inst. Exp. Med. ASCR/Czech Rep.) on distribution of a protein believed to associate with cannabinoid receptors at synapses in the brain. We are planning a study on Myosin 15 with Nathalie Sans/ Mireille Montcouququil (U. Bordeaux, France), related to recent work that they have done with Jonathan Bird (Friedman lab). We have been collaborating with Jennifer Lippincott-Schwartz and Carolyn Ott (now at Janelia) as part of our collaborations with Mark Mattson and Pamela Yao of NIA. We also train/advise/prepare samples for TEM for Kazuyo Takeda (FDA/CBER/Micr. & Imag. Core). We expect that this work will lead to collaborations/publications in near future. We have worked with Maria Rubio (Audi. Res. Gr./U. Pitt.) on deafness effects on auditory synapses in the cochlear nucleus using EM Tomography. We expect that this will continue sometime next year. We also had agreed to a collaboration with Rebecca Seal (Dep. Neurobiol. Otolaryng./U. Pitt.) on synaptic changes in hair cells in VGLUT3 knockout mice. We expect that this will continue sometime next year. Last year, we published research+review papers on NMDA receptor trafficking with Martin Horak of the Czech Republic. We may do some research with his lab next year. In addition to collaborations, Ron Petralia serves as a consultant (since last year) on studies: (T. Fujikawa (Japan)-GluRs in hair cell synapses; H. Xia (LSU)-PP1/I-2; Wenqin Luo (U. Penn.); Y.-X. Tao (NJ Med. Sch./Rutgers)- GluR changes in spinal cord; J. Erickson (LSU)-SNAT8.
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