Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0027819 (neuroblastoma)
 27,800 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Near-field optics (NFO) overcomes the diffraction limit of light microscopes and permits visualization of single molecules. However, despite numerous applications of NFO in the physical sciences, there is still a paucity of applications in the neurosciences. In this work, the authors have developed NFO probes to image intracellular dynamic processes in living cells. This is the first time a NFO probe has been inserted inside a living cell to deliver light to a spatially controlled region for optical measurements and to record cellular responses to external stimuli. Two different optical detection systems (CCD camera and avalanche photon detection) were developed to monitor cellular responses to drug administration in two different cell types. NG108-15 neuroblastoma cells and vascular smooth muscle cells (VSMC) were penetrated with NFO probes. Intracellular Ca2+ increases post drug stimulation were detected by NFO probes. The cells were loaded with either fura-2/AM or fluo-3/AM calcium dyes. VSMC were stimulated with angiotensin II, resulting in a precise area of intracellular Ca2+ increase. Different response profiles of Ca2+ increases were observed after ionomycin and bradykinin administration in NG108-15 cells. Responsive heterogeneities due to ionomycin among different cells of the same type were recorded. The results show that NFO probes make possible real-time visualization of intracellular events. With refinement, intracellular NFO probes offer the potential of probing cell function with fast temporal and excellent spatial resolutions.
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PMID:Probing intracellular dynamics in living cells with near-field optics. 1047 78

A method is described to enable the recording of transient intracellular calcium changes in deep brain structures in anesthetized and awake animals using a fluorescent indicator combined with in vivo optical detection methods. Optrodes were fabricated using a bifurcated fiber-optic cable with an attached infusion guide cannula. After intracranial implantation of an optrode, animals were prepared in the following manner, (1) rats (intra-striatal) and monkeys (intra-putamen) were infused with the fluorescent calcium indicator, Oregon Green, to load intrinsic cells; or (2) rats were intra-striatally transplanted with a slurry of dye-loaded IMR-32 neuroblastoma cells via pipette ejection. Excitation light from an argon-ion laser was launched through the optrode and passed into the tissue. The resulting calcium-induced fluorescence signals were captured by the optrode, then detected and processed by externalized photomultiplier- and CCD-based spectrometer electronics. In approximately 25% of all intrinsic cell recordings, the baseline fluorescence intensity was relatively stable over time whereas in the remainder, large amplitude oscillations were observed with a frequency in the range of 0.5-2 Hz. These Ca(2+) transients were inhibited by local infusion of 10 microM omega-conotoxin MVIIC and 1 microM TTX. Extracellular electrophysiological recordings that were made adjacent to the optrode tip revealed that the Ca(2+) oscillations were in phase with the burst firing of striatal neurons. This suggested that the optical signals had a neuronal origin, most likely from medium spiny neurons. Baseline fluorescence intensity increased during infusion of high [K(+)](o), the calcium ionophore, A-23187, or during temporary bilateral carotid artery occlusion. Monkey (Saimiri sciureus) putamen recordings also affirmed the presence of similar calcium-related transients in a non-human primate. In the transplant preparations, the IMR-32 cells displayed a stable, non-oscillating baseline fluorescence. They were similarly responsive to high [K(+)](o) challenge and appeared viable for at least several hours. Similar optical recording approaches might be applied to monitor other fluorescent, chemiluminescent or bioluminescent events from almost any brain structure. Moreover, transplanted transfected cells expressing a single specific receptor or ion-channel protein may effectively serve as biosensing elements for the measurement of extracellular neurochemical signaling.
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PMID:In vivo spectrometric calcium flux recordings of intrinsic Caudate-Putamen cells and transplanted IMR-32 neuroblastoma cells using miniature fiber optrodes in anesthetized and awake rats and monkeys. 1093 38