期刊
JOURNAL OF NEUROPHYSIOLOGY
卷 99, 期 3, 页码 1535-1544出版社
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/jn.01127.2007
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资金
- NIGMS NIH HHS [R01 GM053395, GM 65473, R01 GM053395-12] Funding Source: Medline
- NIMH NIH HHS [MH 07175-05] Funding Source: Medline
- ONDIEH CDC HHS [ND-S 44564] Funding Source: Medline
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM053395, R24GM065473] Funding Source: NIH RePORTER
To understand the precise microarchitecture of the cortical circuitry, it is crucial to know the distribution of synaptic connections and their synaptic strengths at the level of a single cell, rather than a group of cells. Here, we describe a new application of two-photon photolysis of caged glutamate that enabled us to induce an action potential in only a small number (about five) of pyramidal neurons by increasing the volume of two-photon excitation by reducing the effective numerical aperture of the objective. We performed whole cell patch-clamp recordings from layer 2/3 pyramidal neurons in the rat visual cortex and stimulated many neurons in a large three-dimensional space (similar to 600 x 600 x 100 mu m) including neurons in layers 2/3 and 4 using this new technique. We mapped the density and amplitude of unitary excitatory postsynaptic currents and found that the basic microarchitecture of excitatory synaptic connections consists of two regions: a columnar, dense core region with a radius of 150 mu m and an outer, sparse region. The dense core region includes the majority of strong synaptic connections in layer 2/3. Our results reveal the columnar organization of synaptic connectivity in the rat visual cortex, where functional columns have not been clearly demonstrated. Thus this technique will be a uniquely powerful tool for quantifying synaptic connectivity and manipulating neural activity at the single-cell level.
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