On May 6, 2009, the X-ray imaging and biomedical application beamline(BL13W1) at Shanghai Synchrotron Radiation Facility(SSRF) officially opened to users, with 8–72.5 ke V X-rays. The experimental station is equipped...On May 6, 2009, the X-ray imaging and biomedical application beamline(BL13W1) at Shanghai Synchrotron Radiation Facility(SSRF) officially opened to users, with 8–72.5 ke V X-rays. The experimental station is equipped with four sets of X-ray CCD detectors of different pixel size(0.19–24 μm) for on-line phase-contrast imaging and micro-CT imaging with 0.8 μm spatial resolution and 1 ms temporal resolution. An in vivo microCT experiment for a living insect was realized in 4 s. An X-ray fluorescence detector is equipped for X-ray fluorescence mapping imaging and X-ray fluorescence micro-CT imaging with 50 μm spatial resolution. In order to meet different requirements from the users, several experimental methods, such as X-ray spiral micro-CT, Xray local micro-CT, X-ray fast micro-CT, X-ray grating-based differential micro-CT, X-ray fluorescence microCT and X-ray quantitative micro-CT have been developed, and nearly 60 papers related to those developments for this beamline have been published. Moreover, the beamline has realized the remote fast CT reconstruction,providing a great convenience for the users to process experimental data at their offices. As of August 2014,the beamline has offered the user beamtime of(23 145 h), from which 232 user papers have been published,including 151 SCI papers and 55 papers with SCI impact factor > 3. The quantity and quality of the user paper outcome keep a steady increase. Some typical user experimental results are introduced.展开更多
Objective To explore the possible differential trafficking properties of the dopamine D 1-like receptor subtypes, D 1 receptor and D5 receptor. Methods To visualize distributions of dopamine D 1-like receptor subtypes...Objective To explore the possible differential trafficking properties of the dopamine D 1-like receptor subtypes, D 1 receptor and D5 receptor. Methods To visualize distributions of dopamine D 1-like receptor subtypes at subcellular level, the yellow and cyan variants of green fluorescent protein （GFP） were used to tag D1 and D5 receptors. After transfection with the tagged dopamine receptors, the neuroblastoma cells NG108-15 were treated with D1 agonist SKF38393 or acetylcholine （ACh）. Then we observed the subcellular distributions of the tagged receptors under the confocal microscopy and tried to determine trafficking properties by comparing their distribution patterns before and after the drug treatment. Results In resting conditions, D 1 receptors located in the plasma membrane of NG108-15 cells, while D5 receptors located in both plasma membrane and cytosol. With the pre-treatment of SKF38393, the subcellular distribution of D1 receptors was changed. The yellow particle-like fluorescence of tagged D 1 receptors appeared in the cytosol, indicating that D 1 receptors were internalized into cytosol from the cell surface. Same situation also occurred in ACh pre-treatment. In contrast, the subcellular distribution of D5 receptors was not changed after SKF38393 or ACh treatment, indicating that D5R was not translocated to cell surface. Interestingly, when D1 and D5 receptors were co-expressed in the same cell, both kept their distinct subcellular distribution patterns and the trafficking properties. Conclusion Our present study reveals that in NG108-15 nerve cells, dopamine D1 and D5 receptors exhibit differential subcellular distribution patterns, and only D1 receptor has a marked trafficking response to the drug stimulation. We further discuss the potential role of the differential trafficking properties of D1-like receptors in complex modulation of DA signaling.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.11475248 and 11105213)
文摘On May 6, 2009, the X-ray imaging and biomedical application beamline(BL13W1) at Shanghai Synchrotron Radiation Facility(SSRF) officially opened to users, with 8–72.5 ke V X-rays. The experimental station is equipped with four sets of X-ray CCD detectors of different pixel size(0.19–24 μm) for on-line phase-contrast imaging and micro-CT imaging with 0.8 μm spatial resolution and 1 ms temporal resolution. An in vivo microCT experiment for a living insect was realized in 4 s. An X-ray fluorescence detector is equipped for X-ray fluorescence mapping imaging and X-ray fluorescence micro-CT imaging with 50 μm spatial resolution. In order to meet different requirements from the users, several experimental methods, such as X-ray spiral micro-CT, Xray local micro-CT, X-ray fast micro-CT, X-ray grating-based differential micro-CT, X-ray fluorescence microCT and X-ray quantitative micro-CT have been developed, and nearly 60 papers related to those developments for this beamline have been published. Moreover, the beamline has realized the remote fast CT reconstruction,providing a great convenience for the users to process experimental data at their offices. As of August 2014,the beamline has offered the user beamtime of(23 145 h), from which 232 user papers have been published,including 151 SCI papers and 55 papers with SCI impact factor > 3. The quantity and quality of the user paper outcome keep a steady increase. Some typical user experimental results are introduced.
文摘Objective To explore the possible differential trafficking properties of the dopamine D 1-like receptor subtypes, D 1 receptor and D5 receptor. Methods To visualize distributions of dopamine D 1-like receptor subtypes at subcellular level, the yellow and cyan variants of green fluorescent protein （GFP） were used to tag D1 and D5 receptors. After transfection with the tagged dopamine receptors, the neuroblastoma cells NG108-15 were treated with D1 agonist SKF38393 or acetylcholine （ACh）. Then we observed the subcellular distributions of the tagged receptors under the confocal microscopy and tried to determine trafficking properties by comparing their distribution patterns before and after the drug treatment. Results In resting conditions, D 1 receptors located in the plasma membrane of NG108-15 cells, while D5 receptors located in both plasma membrane and cytosol. With the pre-treatment of SKF38393, the subcellular distribution of D1 receptors was changed. The yellow particle-like fluorescence of tagged D 1 receptors appeared in the cytosol, indicating that D 1 receptors were internalized into cytosol from the cell surface. Same situation also occurred in ACh pre-treatment. In contrast, the subcellular distribution of D5 receptors was not changed after SKF38393 or ACh treatment, indicating that D5R was not translocated to cell surface. Interestingly, when D1 and D5 receptors were co-expressed in the same cell, both kept their distinct subcellular distribution patterns and the trafficking properties. Conclusion Our present study reveals that in NG108-15 nerve cells, dopamine D1 and D5 receptors exhibit differential subcellular distribution patterns, and only D1 receptor has a marked trafficking response to the drug stimulation. We further discuss the potential role of the differential trafficking properties of D1-like receptors in complex modulation of DA signaling.
