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2, and a contrast of 92%. The velocity of the moving pattern depended on the type of experiment and is indicated in the figure legends. Electrophysiological recording: Electrodes were pulled on a Brown-Flaming micropipette puller (P-97, Sutter Instruments) using thin-wall glass capillaries with a diameter of 1 mm (GC100TF-10, Clark). When filled with 2 M KAc, 0.5 M KCl, and 8.8 mM calcium green, they had resistances of about 30 to 40 MΩ. A SEC-10L amplifier (npi-electronics) was used throughout the experiments and was operated in the Bridge or discontinuous current clamp mode. For data analysis, the output signal of the amplifier was fed to a PC 486 via a 12-bit A/D converter (CIO-DAS16, ComputerBoards) at a sampling rate of 3 kHz and stored to hard disc. The motor control unit for the rotating cylinder was controlled by a PC 468. Optical recording: We used an upright epifluorescent microscope (Axiotech Vario, Zeiss) with the fluorescein isothiocyanate filter set 9 from Zeiss (excitation filter, 450 to 490 nm; beam splitter, 510 nm; barrier filter, 516 to 565 nm), an Epiplan × 10/0.20 objective, and a charge-coupled device (CCD) camera (PXL, Photometrics) connected to a Power-Mac (Apple). Images were taken at 1 Hz (Fig. 1) or at 4 Hz (Figs. 3 and 4) at 128 × 128 pixel resolution and were evaluated with the IPLab software (Signal Analytics). The first frame of each image series was taken as the reference frame, which was subtracted from each following image. This resulted in a series of difference images (Δf), which were subsequently divided by the reference frame (Δf/f). The Δf/f time courses shown in Figs. 1, 3, and 4 were obtained by averaging the pixel values in different areas in the Δf/f image series.
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The decrease in calcium concentration during hyperpolarization is indicative of a partial activation of VACCs at the cell's resting potential. This has in fact been directly measured for the CH cells in voltage clamp experiments (15). The calcium current showed little inactivation and its activation characteristic was rather flat, with about 30% of maximum current at resting potential.
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15
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3543124359
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Four different LPTCs were visually stimulated by gratings moving at a constant velocity of 15.6°, 52°, 156°, and 234° per second. Although each of the cells showed differences in response strength, the relationship between membrane potential and dendritic calcium concentration was fairly linear with an average correlation coefficient of 0.92 ± 0.05 (mean ± SEM).
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2. The pattern had a spatial wavelength of 32°, a vertical extent of 64°, and an effective contrast of 3, moving at a constant velocity of 16°/s to result in a temporal frequency of 0.5 Hz.
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Fourier analysis of calcium signals resulted in the following amplitudes relative to the constant response (mean ± SEM; n = 4): 21% ± 4% for the first harmonic and 11% ± 2% for the second harmonic. The corresponding values for the simulated dendritic membrane potentials in Fig. 2 are 18% and 8%, respectively.
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i ratios. However, there was no effect on the cancellation of the local input modulations by dendritic integration, resulting in a smooth axonal signal in each case.
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We are grateful to H. Nguyen for excellent technical assistance and to T. Brotz, J. Haag, and T. Oertner for critically reading the manuscript.
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