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Ca 2+ Na + Presynaptic neuron Postsynaptic neuron Synaptic cleft.

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Presentation on theme: "Ca 2+ Na + Presynaptic neuron Postsynaptic neuron Synaptic cleft."— Presentation transcript:

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2 Ca 2+ Na + Presynaptic neuron Postsynaptic neuron Synaptic cleft

3 Ca 2+ Na + Presynaptic neuron Synaptic cleft Postsynaptic neuron

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5 Calcium indicators Aequorea victoria Ca 2+ binding sites Coelenterazine Colenteramide Shimomura et al., 1962

6 Calcium indicators Fura-2 Grynkiewicz et al., 1985 Ferrari et al., 2012

7 Calcium indicators Yellow Cameleon Calmodulin binding peptide Nagai et al., 2004 Von Der Behrens’ lab

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9 Förster Resonance Energy Transfer (FRET)

10 Calcium indicators Nakai et al., 2001 Chen et al., 2012

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14 Kitamura et al., 2008 “Shadow patching” technique

15 Svoboda et al., 1997

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17 Eilers and Konnerth, 2009

18 Judkewitz et al., 2009

19 Stosiek et al., 2003

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24 Advantages of 2 photon microscopy: 1.2 photons of half the energy  less damage. 2.Image just the focal plane  better z-resolution (and also less damage). 3.No need of a pinhole  more efficient in light collection. Figure 4

25 Murayama M, Pérez-Garci E, Lüscher HR, Larkum ME. Fiberoptic system for recording dendritic calcium signals in layer 5 neocortical pyramidal cells in freely moving rats. J Neurophysiol. (2007) Grienberger C, Adelsberger H, Stroh A, Milos RI, Garaschuk O, Schierloh A, Nelken I, Konnerth A Sound-evoked network calcium transients in mouse auditory cortex in vivo. J Physiol. (2012) Figure 4

26 Sawinski J, Wallace DJ, Greenberg DS, Grossmann S, Denk W, Kerr JN. Visually evoked activity in cortical cells imaged in freely moving animals. PNAS (2009) Figure 4

27 Yuste R and Denk W Dendritic spines as basic functional units of neuronal integration. Nature (1995) Takechi H, Eilers J, Konnerth A A new class of synaptic response involving calcium release in dendritic spines. Nature (1998) Kreitzer AC, Gee KR, Archer EA, Regehr WG Monitoring presynaptic calcium dynamics in projection fibers by in vivo loading of a novel calcium indicator. Neuron (2000) Koester HJ, Sakmann B Calcium dynamics associated with action potentials in single nerve terminals of pyramidal cells in layer 2/3 of the young rat neocortex. J Physiol. (2000) Figure 5

28 Svoboda K, Denk W, Kleinfeld D, Tank DW. In vivo dendritic calcium dynamics in neocortical pyramidal neurons. Nature (1997) Figure 6

29 Chen X, Leischner U, Rochefort NL, Nelken I, Konnerth A. Functional mapping of single spines in cortical neurons in vivo. Nature (2011) 1000 frames per second Figure 6

30 Stosiek C, Garaschuk O, Holthoff K, Konnerth A. In vivo two-photon calcium imaging of neuronal networks. PNAS (2003) Figure 7

31 Ohki K, Chung S, Ch'ng YH, Kara P, Reid RC. Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex. Nature (2005) Figure 7

32 Figure 8 Komiyama T, Sato TR, O'Connor DH, Zhang YX, Huber D, Hooks BM, Gabitto M, Svoboda K. Learning-related fine-scale specificity imaged in motor cortex circuits of behaving mice. Nature (2010)

33 Figure 8 Dombeck DA, Harvey CD, Tian L, Looger LL, Tank DW. Functional imaging of hippocampal place cells at cellular resolution during virtual navigation. Nature Neuroscience (2010)

34 Dmitriy Aronov, David W. Tank Engagement of Neural Circuits Underlying 2D Spatial Navigation in a Rodent Virtual Reality System. Neuron (yesterday)

35 Our Set-up

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