The lab has continued to do some neuron cell culture preparations as needed by these researchers. We also oversee maintenance of the JEOL 1010 transmission electron microscope and related equipment, as well as the Zeiss LSM710 laser confocal microscope and Leica Biowave System, which continued to be used here by some of these researchers, and others from adjacent NIDCD labs, in FY12. Ronald Petralia and Ya-Xian Wang continued on several projects begun in FY11 and which were described in last years annual report, and began some new ones in FY12. These are described in detail in the annual report of the new Advanced Imaging Core. However, we have included the associated published works here also in this report, since most of them were begun in FY11 in this project. Otherwise, we will summarize here specifically the work of Gail Seabold, Kai Chang, and Chan-Ying Zheng. Gail Seabold worked to finish two projects while she was here in FY12. She completed the revisions of her major paper on the synaptic adhesion-like molecule, SALM1, and the role of dileucine and PDZ-binding motifs in its trafficking in hippocampal neurons. This work was a collaboration with Ronald Petralia, Robert Wenthold, and Ya-Xian Wang, along with former members of the laboratory, Philip Wang and Kai Chang. It finally was published in the Journal of Biological Chemistry in early 2012, and was featured for the cover photo of the issue. She also worked with Ronald Petralia (and Martin Horak) on a large review paper on glutamate receptor trafficking and plasticity that was published on-line in the Spring of 2012. Gail also worked with Ronald Petralia on some preliminary new studies that are in progress. In addition, Gail was of great benefit in the laboratory, helping to organize and cull through antibodies, DNA plasmids, and other materials in the laboratory, as well as helping out in the laboratory in general. Kai Chang has used techniques of molecular biology, cell biology and histology, to characterize melanomas from several strains of transgenic mice for the metabotropic glutamate receptor, mGluR5 (Choi K.Y and Chang K., et al. PNAS 108 (37): 15219-15224, 2011). In the next phase of study on mGluR5-induced melanoma, he focused on the molecular mechanisms that lead to tumorigenesis of melanocytes by mGluR5 ectopic expression. His major accomplishments in this include: unraveled the signal pathways of ERK1/2 activation in mGluR5-transfected melanocytes, characterized proteins involved in melanomagenesis induced by mGluR5 (potentially useful in diagnosis and therapy), elucidated the function of different mGluR5 domains and showed that ectodomain shedding was associated with cell proliferation, established a melan-a transforming assay and stable melan-a cell lines expressing mGluRs, useful for in vitro tumorigenesis assays, and studied non-receptor tyrosine kinases that may bind to the C-tail of mGluRs and function as co-receptors. Chan-Ying Zheng had three papers with Ronald Petralia and Ya-Xian Wang during 2011, including 2 research papers published in collaboration with Drs. Wenthold and Kachar. These were follow-up papers to one published by these three in 2010. One was a description of fluorescence recovery after photobleaching of pEGFP vector in spines of cultured hippocampal neurons, published in the Journal of Visual Experiments. Another was a study on super resolution microscopy that revealed the slightly different localization of the MAGUKs (synaptic scaffolding proteins), SAP102 and PSD-95 a difference that required super resolution to identify. The third was a review article on the scaffolding proteins, MAGUKs, and their role in synaptic development and plasticity. This was published in the Neuroscientist, and also was used for the cover photo for that issue. After Chan-Ying joined the laboratory of Dr. Katherine Roche in NINDS, she began studying the trafficking and function of two AMPA receptor binding proteins, Gamma 8 and Cornichon 2. These projects are ongoing. Recently, she received two knockout (KO) mice. The Gamma 2 KO mice were purchased from Jackson Laboratories and Gamma 8 KO mice were gifts from Dr. Roger Nicolls group. Chan-Ying is working on Gamma 8 and Cornichon 2 projects using imaging and biochemistry methods. She recently contributed to a Cornichon 2 paper, which has been submitted by the Nicoll group last month. She is the third author of that paper. Also this year, Chan-Ying has been actively reviewing papers submitted to journals by other research groups. She has reviewed 10 manuscripts since November 2011. Half of them were referred by Ronald Petralia, who helped to review her comments in her reviews. The journals that she reviewed include: 2011-- Journal of Molecular Cell Biology, Science China, Frontiers in Biology. 2012-Plos One*(2 papers), Communicative &Integrative Biology, Neuropharmacology, International Journal of Developmental Neuroscience*(2 papers), Acta Pharmacologica Sinica. Chan-Ying will continue her work with Katherine Roche into FY13.
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