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Mapping inhibitory neuronal circuits by laser scanning photostimulation.
J Vis Exp 2011; (56)JV

Abstract

Inhibitory neurons are crucial to cortical function. They comprise about 20% of the entire cortical neuronal population and can be further subdivided into diverse subtypes based on their immunochemical, morphological, and physiological properties. Although previous research has revealed much about intrinsic properties of individual types of inhibitory neurons, knowledge about their local circuit connections is still relatively limited. Given that each individual neuron's function is shaped by its excitatory and inhibitory synaptic input within cortical circuits, we have been using laser scanning photostimulation (LSPS) to map local circuit connections to specific inhibitory cell types. Compared to conventional electrical stimulation or glutamate puff stimulation, LSPS has unique advantages allowing for extensive mapping and quantitative analysis of local functional inputs to individually recorded neurons. Laser photostimulation via glutamate uncaging selectively activates neurons perisomatically, without activating axons of passage or distal dendrites, which ensures a sub-laminar mapping resolution. The sensitivity and efficiency of LSPS for mapping inputs from many stimulation sites over a large region are well suited for cortical circuit analysis. Here we introduce the technique of LSPS combined with whole-cell patch clamping for local inhibitory circuit mapping. Targeted recordings of specific inhibitory cell types are facilitated by use of transgenic mice expressing green fluorescent proteins (GFP) in limited inhibitory neuron populations in the cortex, which enables consistent sampling of the targeted cell types and unambiguous identification of the cell types recorded. As for LSPS mapping, we outline the system instrumentation, describe the experimental procedure and data acquisition, and present examples of circuit mapping in mouse primary somatosensory cortex. As illustrated in our experiments, caged glutamate is activated in a spatially restricted region of the brain slice by UV laser photolysis; simultaneous voltage-clamp recordings allow detection of photostimulation-evoked synaptic responses. Maps of either excitatory or inhibitory synaptic input to the targeted neuron are generated by scanning the laser beam to stimulate hundreds of potential presynaptic sites. Thus, LSPS enables the construction of detailed maps of synaptic inputs impinging onto specific types of inhibitory neurons through repeated experiments. Taken together, the photostimulation-based technique offers neuroscientists a powerful tool for determining the functional organization of local cortical circuits.

Authors+Show Affiliations

Department of Anatomy and Neurobiology, School of Medicine, University of California, Irvine, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Video-Audio Media

Language

eng

PubMed ID

22006064

Citation

Ikrar, Taruna, et al. "Mapping Inhibitory Neuronal Circuits By Laser Scanning Photostimulation." Journal of Visualized Experiments : JoVE, 2011.
Ikrar T, Olivas ND, Shi Y, et al. Mapping inhibitory neuronal circuits by laser scanning photostimulation. J Vis Exp. 2011.
Ikrar, T., Olivas, N. D., Shi, Y., & Xu, X. (2011). Mapping inhibitory neuronal circuits by laser scanning photostimulation. Journal of Visualized Experiments : JoVE, (56), doi:10.3791/3109.
Ikrar T, et al. Mapping Inhibitory Neuronal Circuits By Laser Scanning Photostimulation. J Vis Exp. 2011 Oct 6;(56) PubMed PMID: 22006064.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mapping inhibitory neuronal circuits by laser scanning photostimulation. AU - Ikrar,Taruna, AU - Olivas,Nicholas D, AU - Shi,Yulin, AU - Xu,Xiangmin, Y1 - 2011/10/06/ PY - 2011/10/19/entrez PY - 2011/10/19/pubmed PY - 2012/5/19/medline JF - Journal of visualized experiments : JoVE JO - J Vis Exp IS - 56 N2 - Inhibitory neurons are crucial to cortical function. They comprise about 20% of the entire cortical neuronal population and can be further subdivided into diverse subtypes based on their immunochemical, morphological, and physiological properties. Although previous research has revealed much about intrinsic properties of individual types of inhibitory neurons, knowledge about their local circuit connections is still relatively limited. Given that each individual neuron's function is shaped by its excitatory and inhibitory synaptic input within cortical circuits, we have been using laser scanning photostimulation (LSPS) to map local circuit connections to specific inhibitory cell types. Compared to conventional electrical stimulation or glutamate puff stimulation, LSPS has unique advantages allowing for extensive mapping and quantitative analysis of local functional inputs to individually recorded neurons. Laser photostimulation via glutamate uncaging selectively activates neurons perisomatically, without activating axons of passage or distal dendrites, which ensures a sub-laminar mapping resolution. The sensitivity and efficiency of LSPS for mapping inputs from many stimulation sites over a large region are well suited for cortical circuit analysis. Here we introduce the technique of LSPS combined with whole-cell patch clamping for local inhibitory circuit mapping. Targeted recordings of specific inhibitory cell types are facilitated by use of transgenic mice expressing green fluorescent proteins (GFP) in limited inhibitory neuron populations in the cortex, which enables consistent sampling of the targeted cell types and unambiguous identification of the cell types recorded. As for LSPS mapping, we outline the system instrumentation, describe the experimental procedure and data acquisition, and present examples of circuit mapping in mouse primary somatosensory cortex. As illustrated in our experiments, caged glutamate is activated in a spatially restricted region of the brain slice by UV laser photolysis; simultaneous voltage-clamp recordings allow detection of photostimulation-evoked synaptic responses. Maps of either excitatory or inhibitory synaptic input to the targeted neuron are generated by scanning the laser beam to stimulate hundreds of potential presynaptic sites. Thus, LSPS enables the construction of detailed maps of synaptic inputs impinging onto specific types of inhibitory neurons through repeated experiments. Taken together, the photostimulation-based technique offers neuroscientists a powerful tool for determining the functional organization of local cortical circuits. SN - 1940-087X UR - https://www.unboundmedicine.com/medline/citation/22006064/Mapping_inhibitory_neuronal_circuits_by_laser_scanning_photostimulation_ L2 - https://dx.doi.org/10.3791/3109 DB - PRIME DP - Unbound Medicine ER -