@article {46744, title = {Visualization and neuronal cell targeting during electrophysiological recordings facilitated by quantum dots.}, journal = {Proceedings of SPIE}, volume = {9305}, year = {2015}, month = {2015 Mar 10}, pages = {9305Y}, abstract = {

The simultaneous visualization, identification and targeting of neurons during patch clamp-mediated electrophysiological recordings is a basic technique in neuroscience, yet it is often complicated by the inability to visualize the pipette tip, particularly in deep brain tissue. Here we demonstrate a novel approach in which fluorescent quantum dot probes are used to coat pipettes prior to their use. The strong two-photon absorption cross sections of the quantum dots afford robust contrast at significantly deeper penetration depths than current methods allow. We demonstrate the utility of this technique in multiple recording formats both in vitro and in vivo where imaging of the pipettes is achieved at remarkable depths (up to 800 microns). Notably, minimal perturbation of cellular physiology is observed over the hours-long time course of neuronal recordings. We discuss our results within the context of the role that quantum dot nanoprobes may play in understanding neuronal cell physiology.

}, doi = {10.1117/12.2076934}, author = {Field, Lauren D and Andrasfalvy, Bertalan K and Gali{\~n}anes, Gregorio L and Huber, Daniel and Barbic, Mladen and Macklin, John J and Susumu, Kimihiro and Delehanty, James B and Huston, Alan L and Makara, Judit K and Medintz, Igor L} } @article {45812, title = {Quantum dot-based multiphoton fluorescent pipettes for targeted neuronal electrophysiology.}, journal = {Nature Methods}, volume = {11}, year = {2014}, month = {2014 Dec}, pages = {1237-41}, abstract = {

Targeting visually identified neurons for electrophysiological recording is a fundamental neuroscience technique; however, its potential is hampered by poor visualization of pipette tips in deep brain tissue. We describe quantum dot-coated glass pipettes that provide strong two-photon contrast at deeper penetration depths than those achievable with current methods. We demonstrated the pipettes{\textquoteright} utility in targeted patch-clamp recording experiments and single-cell electroporation of identified rat and mouse neurons in vitro and in vivo.

}, issn = {1548-7105}, doi = {10.1038/nmeth.3146}, author = {Andrasfalvy, Bertalan K and Gali{\~n}anes, Gregorio L and Huber, Daniel and Barbic, Mladen and Macklin, John J and Susumu, Kimihiro and Delehanty, James B and Huston, Alan L and Makara, Judit K and Medintz, Igor L} } @article {5923, title = {Ion channel gradients in the apical tuft region of CA1 pyramidal neurons.}, journal = {PLoS one}, volume = {7}, year = {2012}, month = {2012}, pages = {e46652}, abstract = {

Dendritic ion channels play a critical role in shaping synaptic input and are fundamentally important for synaptic integration and plasticity. In the hippocampal region CA1, somato-dendritic gradients of AMPA receptors and the hyperpolarization-activated cation conductance (I(h)) counteract the effects of dendritic filtering on the amplitude, time-course, and temporal integration of distal Schaffer collateral (SC) synaptic inputs within stratum radiatum (SR). While ion channel gradients in CA1 distal apical trunk dendrites within SR have been well characterized, little is known about the patterns of ion channel expression in the distal apical tuft dendrites within stratum lacunosum moleculare (SLM) that receive distinct input from the entorhinal cortex via perforant path (PP) axons. Here, we measured local ion channels densities within these distal apical tuft dendrites to determine if the somato-dendritic gradients of I(h) and AMPA receptors extend into distal tuft dendrites. We also determined the densities of voltage-gated sodium channels and NMDA receptors. We found that the densities of AMPA receptors, I(h,) and voltage-gated sodium channels are similar in tuft dendrites in SLM when compared with distal apical dendrites in SR, while the ratio of NMDA receptors to AMPA receptors increases in tuft dendrites relative to distal apical dendrites within SR. These data indicate that the somato-dendritic gradients of I(h) and AMPA receptors in apical dendrites do not extend into the distal tuft, and the relative densities of voltage-gated sodium channels and NMDA receptors are poised to support nonlinear integration of correlated SC and PP input.

}, issn = {1932-6203}, doi = {10.1371/journal.pone.0046652}, author = {Bittner, Katie C and Andrasfalvy, Bertalan K and Magee, Jeffrey C} } @article {4284, title = {Two-photon single-cell optogenetic control of neuronal activity by sculpted light.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {107}, year = {2010}, month = {2010 Jun 29}, pages = {11981-6}, abstract = {

Recent advances in optogenetic techniques have generated new tools for controlling neuronal activity, with a wide range of neuroscience applications. The most commonly used approach has been the optical activation of the light-gated ion channel channelrhodopsin-2 (ChR2). However, targeted single-cell-level optogenetic activation with temporal precessions comparable to the spike timing remained challenging. Here we report fast (\< or = 1 ms), selective, and targeted control of neuronal activity with single-cell resolution in hippocampal slices. Using temporally focused laser pulses (TEFO) for which the axial beam profile can be controlled independently of its lateral distribution, large numbers of channels on individual neurons can be excited simultaneously, leading to strong (up to 15 mV) and fast (\< or = 1 ms) depolarizations. Furthermore, we demonstrated selective activation of cellular compartments, such as dendrites and large presynaptic terminals, at depths up to 150 microm. The demonstrated spatiotemporal resolution and the selectivity provided by TEFO allow manipulation of neuronal activity, with a large number of applications in studies of neuronal microcircuit function in vitro and in vivo.

}, keywords = {Action Potentials, Animals, Cell Line, Dendrites, Electrophysiological Phenomena, Hippocampus, Humans, Lasers, Mice, Nerve Tissue Proteins, Neurons, Optical Phenomena, Patch-Clamp Techniques, Photic Stimulation, Photons, Presynaptic Terminals, Rats, Recombinant Fusion Proteins, Rhodopsin, Sprague-Dawley, Transgenic}, issn = {1091-6490}, doi = {10.1073/pnas.1006620107}, author = {Andrasfalvy, Bertalan K and Zemelman, Boris V and Tang, Jianyong and Vaziri, Alipasha} } @article {2134, title = {Changes in AMPA receptor currents following LTP induction on rat CA1 pyramidal neurones.}, journal = {The Journal of Physiology}, volume = {559}, year = {2004}, month = {2004 Sep 1}, pages = {543-54}, abstract = {

In the CA1 region of the hippocampus, LTP is thought to be initiated by a transient activation of NMDA receptors and is expressed as a persistent increase in synaptic transmission through AMPA receptors. To investigate the postsynaptic modifications of AMPA receptors involved in this enhanced synaptic transmission, the channel density and single-channel properties of extrasynaptic AMPA receptors located in synaptically active dendritic regions were examined following the induction of LTP. Following tetanic stimulation an outside-out patch was excised from the apical dendrite near the point of stimulation and saturating concentrations of glutamate were rapidly applied to the patch. AMPA current amplitude and duration were increased significantly in patches pulled from dendrites that expressed LTP. Non-stationary fluctuation analysis of AMPA currents indicated that AMPA channel number was nearly twofold larger than in controls, while single channel conductance and maximum open-probability were unchanged. Furthermore, while subtle changes in AMPA channel kinetics could also be observed, we did not find any evidence that receptor affinity or rectification properties were altered by LTP induction. Very similar results were found when CaMK-II activity was increased through the intracellular application of Ca/CaM. Together, we interpret our data to indicate that the stimuli used here produce an increased delivery of AMPA receptors to synaptically active regions of the apical dendrite without inducing any significant changes in their basic biophysical properties and that such delivery is a key element in this form of synaptic plasticity.

}, keywords = {AMPA, Animals, Calcium-Calmodulin-Dependent Protein Kinase Type 2, Calcium-Calmodulin-Dependent Protein Kinases, Glutamic Acid, Long-Term Potentiation, Pyramidal Cells, Rats, Receptors, Sprague-Dawley}, issn = {0022-3751}, doi = {10.1002/cbic.201000254}, author = {Andrasfalvy, Bertalan K and Magee, Jeffrey C} } @article {2137, title = {Impaired regulation of synaptic strength in hippocampal neurons from GluR1-deficient mice.}, journal = {The Journal of Physiology}, volume = {552}, year = {2003}, month = {2003 Oct 1}, pages = {35-45}, abstract = {

Neurons of the central nervous system (CNS) exhibit a variety of forms of synaptic plasticity, including associative long-term potentiation and depression (LTP/D), homeostatic activity-dependent scaling and distance-dependent scaling. Regulation of synaptic neurotransmitter receptors is currently thought to be a common mechanism amongst many of these forms of plasticity. In fact, glutamate receptor 1 (GluR1 or GluRA) subunits containing L-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors have been shown to be required for several forms of hippocampal LTP and a particular hippocampal-dependent learning task. Because of this importance in associative plasticity, we sought to examine the role of these receptors in other forms of synaptic plasticity in the hippocampus. To do so, we recorded from the apical dendrites of hippocampal CA1 pyramidal neurons in mice lacking the GluR1 subunit (GluR1 -/-). Here we report data from outside-out patches that indicate GluR1-containing receptors are essential to the extrasynaptic population of AMPA receptors, as this pool was nearly empty in the GluR1 -/- mice. Additionally, these receptors appear to be a significant component of the synaptic glutamate receptor pool because the amplitude of spontaneous synaptic currents recorded at the site of input and synaptic AMPA receptor currents evoked by focal glutamate uncaging were both substantially reduced in these mice. Interestingly, the impact on synaptic weight was greatest at distant synapses such that the normal distance-dependent synaptic scaling used by these cells to counter dendritic attenuation was lacking in GluR1 -/- mice. Together the data suggest that the highly regulated movement of GluR1-containing AMPA receptors between extrasynaptic and synaptic receptor pools is critically involved in establishing two functionally diverse forms of synaptic plasticity: LTP and distance-dependent scaling.

}, keywords = {AMPA, Animals, Hippocampus, Inbred C57BL, Kinetics, Long-Term Potentiation, Mice, Mutant Strains, Neuronal Plasticity, Patch-Clamp Techniques, Pyramidal Cells, Receptors, Synapses}, issn = {0022-3751}, doi = {10.1002/cbic.201000254}, author = {Andrasfalvy, Bertalan K and Smith, Mark A and Borchardt, Thilo and Sprengel, Rolf and Magee, Jeffrey C} }