Two-photon photostimulation and imaging of neural circuits

Nikolenko, Volodymyr; Poskanzer, Kira E.; Yuste, Rafael

doi:10.1038/nmeth1105

Cited by 238 publications

(226 citation statements)

References 28 publications

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“…These widely studied structures were not previously accessible using high-resolution optical techniques. By providing optical access to deep fissures, the methods presented herein open up these structures for study at cellular resolution in behaving animals using the powerful, rapidly expanding palette of optical tools for perturbing and measuring network structure and function (39,40). Finally, with further optimization, our current method can be used to image not only the mPFC and MEC, but also other brain regions within fissures, such as the parasubiculum and retrosplenial granular cortex.…”

Section: Discussionmentioning

confidence: 99%

Cellular resolution optical access to brain regions in fissures: Imaging medial prefrontal cortex and grid cells in entorhinal cortex

Low

Tank

2014

Proc. Natl. Acad. Sci. U.S.A.

114

153

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In vivo two-photon microscopy provides the foundation for an array of powerful techniques for optically measuring and perturbing neural circuits. However, challenging tissue properties and geometry have prevented high-resolution optical access to regions situated within deep fissures. These regions include the medial prefrontal and medial entorhinal cortex (mPFC and MEC), which are of broad scientific and clinical interest. Here, we present a method for in vivo, subcellular resolution optical access to the mPFC and MEC using microprisms inserted into the fissures. We chronically imaged the mPFC and MEC in mice running on a spherical treadmill, using two-photon laser-scanning microscopy and genetically encoded calcium indicators to measure network activity. In the MEC, we imaged grid cells, a widely studied cell type essential to memory and spatial information processing. These cells exhibited spatially modulated activity during navigation in a virtual reality environment. This method should be extendable to other brain regions situated within deep fissures, and opens up these regions for study at cellular resolution in behaving animals using a rapidly expanding palette of optical tools for perturbing and measuring network structure and function.two-photon imaging | medial prefrontal cortex | medial entorhinal cortex | grid cell O ptical tools provide powerful means to measure and perturb the structure and function of neural circuits (1). However, a key set of brain regions remains outside the reach of cellularresolution optical methods: those situated within deep fissures. In the rodent brain, these areas include the medial prefrontal and medial entorhinal cortex (mPFC and MEC), two of the most widely studied regions supporting cognitive functions. The mPFC is centrally important to planning, executive function, learning, and memory (2, 3). Understanding how the mPFC implements these functions at a mechanistic, circuit level remains a longstanding goal of the field. The entorhinal cortex forms the interface between the neocortex and hippocampus and is believed to play an essential role in episodic memory and spatial information processing. These functions are particularly associated with MEC grid cells, which fire on a hexagonal lattice as an animal moves through an environment (4). Grid cells have galvanized a large body of work investigating their computational role and the mechanisms underlying grid formation, which remain important, open questions (4, 5). Moreover, mPFC or MEC dysfunction is associated with addiction, depression, schizophrenia, Alzheimer's disease, and epilepsy.The study of the mPFC, MEC, and neighboring regions could be greatly advanced by in vivo, high-resolution optical tools. However, tissue properties and geometry present significant challenges to deploying these tools within deep fissures. For example, the depth of two-photon microscopy is limited by scattering and out-of-plane excitation (6). Access in the mammalian brain is therefore typically restricted to superficial regions (<500 ...

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Section: Discussionmentioning

confidence: 99%

Cellular resolution optical access to brain regions in fissures: Imaging medial prefrontal cortex and grid cells in entorhinal cortex

Low

Tank

2014

Proc. Natl. Acad. Sci. U.S.A.

114

153

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“…LSPS combined with whole cell recordings is an effective method for mapping local circuit inputs to single neurons, as the simultaneous recording from a postsynaptic neuron with photostimulation of clusters of presynaptic neurons at many different locations provides quantitative measures of spatial distribution of excitatory and inhibitory inputs impinging onto single recorded neurons (Callaway & Katz, 1993;Schubert et al, 2003;Shepherd & Svoboda, 2005;Xu & Callaway, 2009). LSPS has also been combined two-photon calcium imaging to generate detailed functional maps of inputs to individual cells with single-cell and three-dimensional precision (Nikolenko et al, 2007). Different from the aforementioned approaches, our newly developed technique is intended to assess circuit activation and network connectivity at the neuronal population level through fast VSD imaging and photostimulation.…”

Section: Discussionmentioning

confidence: 99%

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

Xu¹

2011

Laser Scanning, Theory and Applications

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“…This results in the release of free neurotransmitter, which will activate synapses in the vicinity of the uncaging location (Petitt et al, 1997). The glutamate uncaging method has already been shown to be useful for single neuron input activation (Nikolenko et al, 2007).…”

Section: Optical Methods and Scalingmentioning

confidence: 99%