I Did Not Realise That!: Top Fifteen BMS-754807 Of This Year

Figure 1��figure supplement 1A). Calcium imaging analysis YC2.60-expressing PNs were visually identified either by high-average somatic fluorescence or high-somatic fluorescence CV, which captures relatively quiescent neurons with intermittent, sparse spiking activity and whose average somatic fluorescence remained low. Boxcar regions of interest (ROIs) were manually drawn around the somatic region BLZ945 in vivo of labeled neurons for which the nucleus was clearly visible within the cross-sectional somatic area. The boxcar was aligned with the outer perimeter of the neuron, and all pixels within the boxcar were taken for analysis. For YC2.60, the ratio, R, of cpVenus fluorescence to ECFP fluorescence was calculated for each ROI and each frame. The ratio measurement requires neuronal signals to be anti-correlated in the two wavelength bands, which allows for easy identification of non-signal artifact, for example, from small animal movements. A continuous function of fluorescence was calculated as ?R/R = (RROI ? R0)/R0, where RROI denotes the average fluorescence ratio within the ROI. The baseline fluorescence ratio, R0, was defined as the median of R. ?R/R was low-pass filtered (Kampa et al., 2011) (3��t; symmetrical Gaussian kernel), and the instantaneous firing rate estimate, ��, in arbitrary units was obtained for each ROI using fast, non-negative PFKM deconvolution (Vogelstein et al., 2010) (OOPSI package, Matlab, MathWorks Inc., Natick, MA). The decay time constant of the non-negative deconvolution for YC2.60 was ?1�C1.5 s, as estimated from simultaneous cell-attached recording and 2-PI (n = 8 neurons). This value was also obtained by minimizing the summed square error between R and a reconstructed R��, made IGF-1R inhibitor by convolving an exponential decaying kernel with �� over the range of �� = ��t ? 15 �� ��t for neuronal populations recorded in vivo and in vitro (Figure 1��figure supplement 1C) and is in line with previous reports (Yamada et al., 2011). The a priori rate estimate for spiking was set to 1 Hz based on a meta-study (Barth and Poulet, 2012), and lower rates were examined systematically in the form of thresholding on �� (�� > ��thr). The standard deviation parameter was estimated for each ?R/R using calcium amplitudes less than 5% ?R/R, corresponding to the average amplitude for single spike detection of YC2.60. Deconvolution parameters were determined, first, from loose-patch recordings (Figure 1��figure supplement 1A,B) and second, from minimizing the residual error in reconstructed ��R/R traces from �� (Figure 1��figure supplement 1C). Both methods yielded an optimal deconvolution time constant of ?1.5 s, in line with previous reports (Yamada et al., 2011). ROIs with ��avg