High Precision and Fast Functional Mapping of Brain Circuitry through Laser Scanning Photostimulation and Fast Dye Imaging

Xu, Xiangmin

doi:10.5772/14943

Cited by 5 publications

(8 citation statements)

References 37 publications

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“…The VSD imaging can monitor neuronal activity in a large area simultaneously, with changes in optical signals closely correlating with membrane potential depolarization of neuronal ensembles (Xu et al 2010;Xu 2011). During VSD imaging experiments, living hippocampal slices were stained with VSD; the CA1 region was stimulated with either electrical stimulation or laser photostimulation, and circuit activation directly visualized and measured by fast VSD imaging ( Fig.…”

Section: Resultsmentioning

confidence: 99%

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Cell-type specific inactivation of hippocampal CA1 disrupts location-dependent object recognition in the mouse

et al. 2013

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The allatostatin receptor (AlstR)/ligand inactivation system enables potent regulation of neuronal circuit activity. To examine how different cell types participate in memory formation, we have used this system through Cre-directed, celltype specific expression in mouse hippocampal CA1 in vivo and examined functional effects of inactivation of excitatory vs. inhibitory neurons on memory formation. We chose to use a hippocampus-dependent behavioral task involving location-dependent object recognition (LOR). The double transgenic mice, with the AlstRs selectively expressed in excitatory pyramidal neurons or inhibitory interneurons, were cannulated, targeting dorsal hippocampus to allow the infusion of the receptor ligand (the allatostatin [AL] peptide) in a time dependent manner. Compared to control animals, AL-infused animals showed no long-term memory for object location. While inactivation of excitatory or inhibitory neurons produced opposite effects on hippocampal circuit activity in vitro, the effects in vivo were similar. Both types of inactivation experiments resulted in mice exhibiting no long-term memory for object location. Together, these results demonstrate that the Cre-directed, AlstR-based system is a powerful tool for cell-type specific manipulations in a behaving animal and suggest that activity of either excitatory neurons or inhibitory interneurons is essential for proper long-term object location memory formation.

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Section: Resultsmentioning

confidence: 99%

“…Our overall imaging system was described previously (Xu et al 2010;Xu 2011). The solution was fed into the slice recording chamber through a pressure-driven flow system with pressurized 95% O 2 -5% CO 2 with a perfusion flow rate of about 2 mL/ min.…”

Section: Slice Imaging Experimentsmentioning

confidence: 99%

Cell-type specific inactivation of hippocampal CA1 disrupts location-dependent object recognition in the mouse

et al. 2013

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“…Our overall system of electrophysiological recording, photostimulation, and imaging was described previously (Xu et al, 2010 ; Xu, 2011 ). Electrophysiological experiments were conducted at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Distinct physiological and developmental properties of hippocampal CA2 subfield revealed by using anti‐Purkinje cell protein 4 (PCP4) immunostaining

Antonio

Liban

Ikrar

et al. 2014

J of Comparative Neurology

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The hippocampal CA2 subfield was initially identified by Lorente de Nó as an anatomically distinct region based on its cytoarchitectural features. While there is an enormous body of literature on other hippocampal subfields (CA1 and CA3), relatively little is known about the physiological and developmental properties of CA2. Here we report identification of the CA2 region in the mouse by immunostaining with a Purkinje cell protein 4 (PCP4) antibody, which effectively delineates CA3/CA2 and CA2/CA1 borders and agrees well with previous cytoarchitectural definitions of CA2. The PCP4-immunostaining delineated CA2 neurons have distinguishable differences in cell morphology, physiology and synaptic circuit connections as compared to distal CA3 and proximal CA1 regions. The average somatic sizes of excitatory cells differ across CA1-3, with the smallest to largest somatic size being CA1

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“…The slices were prepared in a cutting solution (containing in mM: 92 NaCl, 2.5 KCl, 30 NaHCO 3 , 0. Our overall system of electrophysiological recording, photostimulation, and imaging was described previously (Xu 2011;Xu et al 2010). Electrophysiological recordings and photostimulation were similarly performed as in references (Kuhlman et al 2013;Shi et al 2010;Xu et al 2010).…”

Section: Methodsmentioning

confidence: 99%

High-resolution and cell-type-specific photostimulation mapping shows weak excitatory vs. strong inhibitory inputs in the bed nucleus of the stria terminalis

Ikrar

Sun

et al. 2016

Journal of Neurophysiology

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Xu X, Ikrar T, Sun Y, Santos R, Holmes TC, Francesconi W, Berton F. High-resolution and cell-type-specific photostimulation mapping shows weak excitatory vs. strong inhibitory inputs in the bed nucleus of the stria terminalis. J Neurophysiol 115: 3204 -3216, 2016. First published April 6, 2016; doi:10.1152/jn.01148.2015.-The bed nucleus of the stria terminalis (BNST) is a key component of the extended amygdala and has been implicated in anxiety and addiction. As individual neurons function within neural circuits, it is important to understand local microcircuits and larger network connections of identified neuronal types and understand how maladaptive changes in the BNST neural networks are induced by stress and drug abuse. However, due to limitations of classic anatomical and physiological methods, the local circuit organization of synaptic inputs to specific BNST neuron types is not well understood. In this study, we report on the application of high-resolution and cell-type-specific photostimulation methodology developed in our laboratory to local circuit mapping in the BNST. Under calibrated experimental conditions, laser photostimulation via glutamate uncaging or channelrhodopsin-2 photoactivation evokes spiking of BNST neurons perisomatically, without activating spikes from axons of passage or distal dendrites. Whole cell recordings, combined with spatially restricted photostimulation of presynaptic neurons at many different locations over a large region, allow high-resolution mapping of presynaptic input sources to single recorded neurons in the BNST. We constructed maps of synaptic inputs impinging onto corticotrophin-releasing hormone-expressing (CRHϩ) BNST neurons in the dorsolateral BNST and found that the CRHϩ neurons receive predominant local inhibitory synaptic connections with very weak excitatory connections. Through cell-typespecific optogenetic stimulation mapping, we generated maps of somatostatin-expressing neuron-specific inhibitory inputs to BNST neurons. Taken together, the photostimulation-based techniques offer us powerful tools for determining the functional organization of local circuits of specific BNST neuron types. extended amygdala; synaptic connections; glutamatergic; GABAergic; glutamate uncaging; optogenetic stimulation NEW & NOTEWORTHY In this study, we have applied photostimulation-based approaches to local bed nucleus of the stria terminalis circuit mapping of defined neuronal types. We performed physiological and anatomical calibrations for laser scanning photostimulation via glutamate uncaging or channelrhodopsin-2-mediated optogenetic stimulation, to establish experimental conditions allowing for the accuracy of synaptic input mapping analysis. This work will help provide a template for other investigators who wish to apply these techniques to functional mapping of nuclei and other nonlayered brain structures.THE BED NUCLEUS OF THE STRIA TERMINALIS (BNST) is a relatively small structure in the basal forebrain, but it is a major component of the extended amygdala which shar...

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High Precision and Fast Functional Mapping of Brain Circuitry through Laser Scanning Photostimulation and Fast Dye Imaging

Cited by 5 publications

References 37 publications

Cell-type specific inactivation of hippocampal CA1 disrupts location-dependent object recognition in the mouse

Cell-type specific inactivation of hippocampal CA1 disrupts location-dependent object recognition in the mouse

Distinct physiological and developmental properties of hippocampal CA2 subfield revealed by using anti‐Purkinje cell protein 4 (PCP4) immunostaining

High-resolution and cell-type-specific photostimulation mapping shows weak excitatory vs. strong inhibitory inputs in the bed nucleus of the stria terminalis

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