Publications (7)43.04 Total impact
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Article: Characterizing ligand-gated ion channel receptors with genetically encoded Ca2++ sensors.
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ABSTRACT: We present a cell based system and experimental approach to characterize agonist and antagonist selectivity for ligand-gated ion channels (LGIC) by developing sensor cells stably expressing a Ca(2+) permeable LGIC and a genetically encoded Förster (or fluorescence) resonance energy transfer (FRET)-based calcium sensor. In particular, we describe separate lines with human α7 and human α4β2 nicotinic acetylcholine receptors, mouse 5-HT(3A) serotonin receptors and a chimera of human α7/mouse 5-HT(3A) receptors. Complete concentration-response curves for agonists and Schild plots of antagonists were generated from these sensors and the results validate known pharmacology of the receptors tested. Concentration-response relations can be generated from either the initial rate or maximal amplitudes of FRET-signal. Although assaying at a medium throughput level, this pharmacological fluorescence detection technique employs a clonal line for stability and has versatility for screening laboratory generated congeners as agonists or antagonists on multiple subtypes of ligand-gated ion channels. The clonal sensor lines are also compatible with in vivo usage to measure indirectly receptor activation by endogenous neurotransmitters.PLoS ONE 01/2011; 6(1):e16519. · 4.09 Impact Factor -
Article: An in vivo biosensor for neurotransmitter release and in situ receptor activity.
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ABSTRACT: Tools from molecular biology, combined with in vivo optical imaging techniques, provide new mechanisms for noninvasively observing brain processes. Current approaches primarily probe cell-based variables, such as cytosolic calcium or membrane potential, but not cell-to-cell signaling. We devised cell-based neurotransmitter fluorescent engineered reporters (CNiFERs) to address this challenge and monitor in situ neurotransmitter receptor activation. CNiFERs are cultured cells that are engineered to express a chosen metabotropic receptor, use the G(q) protein-coupled receptor cascade to transform receptor activity into a rise in cytosolic [Ca(2+)] and report [Ca(2+)] with a genetically encoded fluorescent Ca(2+) sensor. The initial realization of CNiFERs detected acetylcholine release via activation of M1 muscarinic receptors. We used chronic implantation of M1-CNiFERs in frontal cortex of the adult rat to elucidate the muscarinic action of the atypical neuroleptics clozapine and olanzapine. We found that these drugs potently inhibited in situ muscarinic receptor activity.Nature Neuroscience 12/2009; 13(1):127-32. · 15.53 Impact Factor -
Article: MPScope: a versatile software suite for multiphoton microscopy.
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ABSTRACT: MPScope is a software suite to control and analyze data from custom-built multiphoton laser scanning fluorescence microscopes. The acquisition program MPScan acquires, displays and stores movies, linescans, image stacks or arbitrary regions from up to four imaging channels and up to two analog inputs, while plotting the intensity of regions of interest in real-time. Bidirectional linescans allow 256 x 256 pixel frames to be acquired at up to 10 fps with typical galvanometric scanners. A fast stack mode combines movie acquisition with continuous z-focus motion and adjustment of laser intensity for constant image brightness. Fast stacks can be automated by custom programs running in an integrated scripting environment, allowing a 1 mm(3) cortical volume to be sampled in 1 billion voxels in approximately 1 h. The analysis program MPView allows viewing of stored frames, projections, automatic detection of cells and plotting of their average intensity across frames, direct frame transfer to Matlab, AVI movie creation and file export to ImageJ. The combination of optimized code, multithreading and COM (Common Object Model) technologies enables MPScope to fully take advantage of custom-built two-photon microscopes and to simplify their realization.Journal of Neuroscience Methods 10/2006; 156(1-2):351-9. · 1.98 Impact Factor -
Article: Positive feedback in a brainstem tactile sensorimotor loop.
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ABSTRACT: The trigeminal loop in the brainstem comprises the innermost level of sensorimotor feedback in the rat vibrissa system. Anatomy suggests that this loop relays tactile information from the vibrissae to the motoneurons that control vibrissa movement. We demonstrate, using in vitro and in vivo recordings, that the trigeminal loop consists of excitatory pathways from vibrissa sensory inputs to vibrissa motoneurons in the facial nucleus. We further show that the trigeminal loop implements a rapidly depressing reflex that provides positive sensory feedback to the vibrissa musculature during simulated whisking and contact. On the basis of these findings, we propose that the trigeminal loop provides an enhancement of vibrissa muscle tone upon contact during active touch.Neuron 03/2005; 45(3):447-57. · 14.74 Impact Factor -
Article: Developmental regulation of active and passive membrane properties in rat vibrissa motoneurones.
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ABSTRACT: We characterized the electrophysiological properties of vibrissa motoneurones (vMNs) in rat. Intracellular recordings of vMNs in brainstem slices from animals aged P4 to P5 and P9 to P11, i.e. newborn animals, showed that the subthreshold membrane impedance has the form of passive decay. In particular, the impedance follows the 1/ radical f signature for long dendrites beyond a cut-off frequency of f(c)= 8 Hz. In contrast, the impedance has the form of a resonant filter in vMNs from slices prepared from animals aged P17 to P23, i.e. young animals. The resonance has a peak near 4 Hz and an amplitude of 1.2 times that at low frequencies (f approximately 0.1Hz). The low frequency onset of the resonance is shown to depend on a hyperpolarization-activated depolarizing current, I(h). This current functions as a high-pass filter. The high frequency cut-off of the resonance results from passive decay in long dendrites, similar to the case with newborn animals but with f(c)= 20Hz. In addition to a resonance in subthreshold properties, an enhanced resonance in spiking is observed in young as opposed to newborn animals. The transition from solely passive decay in vMNs from newborn animals to resonance in young animals coincides with the onset of whisking. Further, the width of the resonance encompasses the 4-15Hz range of exploratory whisking. Nonetheless, it remains to be shown if there is a causal relation between the regulation of currents in vMNs and the onset of whisking. In particular, we further observed that the membrane impedance of hypoglossal motoneurones from both newborn and young animals exhibits a subthreshold resonance that also peaks near 4Hz. The amplitude of this resonance increases from 1.1 to 1.4 times that at low frequencies in newborn versus young animals. We conjecture that resonance properties in vibrissa, hypoglossal, and potentially other motoneurones, may serve to transiently and purposely synchronize different orofacial behaviours.The Journal of Physiology 05/2004; 556(Pt 1):203-19. · 4.72 Impact Factor -
Article: e-Phys: a suite of intracellular neurophysiology programs integrating COM (component object model) technologies.
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ABSTRACT: Current computer programs for intracellular recordings often lack advanced data management, are usually incompatible with other applications and are also difficult to adapt to new experiments. We have addressed these shortcomings in e-Phys, a suite of electrophysiology applications for intracellular recordings. The programs in e-Phys use Component Object Model (COM) technologies available in the Microsoft Windows operating system to provide enhanced data storage, increased interoperability between e-Phys and other COM-aware applications, and easy customization of data acquisition and analysis thanks to a script-based integrated programming environment. Data files are extensible, hierarchically organized and integrated in the Windows shell by using the Structured Storage technology. Data transfers to and from other programs are facilitated by implementing the ActiveX Automation standard and distributed COM (DCOM). ActiveX Scripting allows experimenters to write their own event-driven acquisition and analysis programs in the VBScript language from within e-Phys. Scripts can reuse components available from other programs on other machines to create distributed meta-applications. This paper describes the main features of e-Phys and how this package was used to determine the effect of the atypical antipsychotic drug clozapine on synaptic transmission at the neuromuscular junction.Journal of Neuroscience Methods 10/2003; 128(1-2):21-31. · 1.98 Impact Factor -
Article: The enantiomer of strychnine is a weak antagonist of the inhibitory glycine receptor
Bioorganic & Medicinal Chemistry Letters. 5(7):749-752.
Top co-authors
Top Journals
- Journal of Neuroscience Methods (2)
- The Journal of Physiology (1)
- Neuron (1)
- Nature Neuroscience (1)
- PLoS ONE (1)
Institutions
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2003–2009
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University of California, San Diego
- Department of Physics
San Diego, CA, USA
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