Dendritic signals play an essential role in processing visual information in the retina. To study them in neurites too small for electrical recording, we developed an instrument that combines a multi-photon (MP) microscope with a through-the-objective high-resolution visual stimulator. An upright microscope was designed that uses the objective lens for both MP imaging and delivery of visual stimuli to functionally intact retinal explants or eyecup preparations. The stimulator consists of a miniature liquid-crystal-on-silicon display coupled into the optical path of an infrared-excitation laser-scanning microscope. A pair of custom-made dichroic filters allows light from the excitation laser and three spectral bands ('colors') from the stimulator to reach the retina, leaving two intermediate bands for fluorescence imaging. Special optics allow displacement of the stimulator focus relative to the imaging focus. Spatially resolved changes in calcium-indicator fluorescence in response to visual stimuli were recorded in dendrites of different types of mammalian retinal neurons.

Detection of image motion direction begins in the retina, with starburst amacrine cells (SACs) playing a major role. SACs generate larger dendritic Ca2+ signals when motion is from their somata towards their dendritic tips than for motion in the opposite direction. To study the mechanisms underlying the computation of direction selectivity (DS) in SAC dendrites, electrical responses to expanding and contracting circular wave visual stimuli were measured via somatic whole-cell recordings and quantified using Fourier analysis. Fundamental and, especially, harmonic frequency components were larger for expanding stimuli. This DS persists in the presence of GABA and glycine receptor antagonists, suggesting that inhibitory network interactions are not essential. The presence of harmonics indicates nonlinearity, which, as the relationship between harmonic amplitudes and holding potential indicates, is likely due to the activation of voltage-gated channels. [Ca2+] changes in SAC dendrites evoked by voltage steps and monitored by two-photon microscopy suggest that the distal dendrite is tonically depolarized relative to the soma, due in part to resting currents mediated by tonic glutamatergic synaptic input, and that high-voltage–activated Ca2+ channels are active at rest. Supported by compartmental modeling, we conclude that dendritic DS in SACs can be computed by the dendrites themselves, relying on voltage-gated channels and a dendritic voltage gradient, which provides the spatial asymmetry necessary for direction discrimination.

No abstract

No abstract

How direction of image motion is detected as early as at the level of the vertebrate eye has been intensively studied in retina research. Although the first direction-selective (DS) ret­inal ganglion cells were already described in the 1960s and have since then been in the fo­cus of many studies, scientists are still puz­zled by the intricacy of the neuronal circuits and computational mechanisms underlying retinal direction selectivity. The fact that the retina can be easily isolated and studied in a Petri dish-by presenting light stimuli while recording from the various cell types in the retinal circuits-in combination with the ex­tensive anatomical, molecular and physiolog­ical knowledge about this part of the brain presents a unique opportunity for studying this intriguing visual circuit in detail. This ar­ticle provides a brief overview of the histo­ry of research on retinal direction selectivi­ty, but then focuses on the past decade and the progress achieved, in particular driven by methodological advances in optical record­ing techniques, molecular genetics approach­es and large-scale ultrastructural reconstruc­tions. As it turns out, retinal direction selec­tivity is a complex, multi-tiered computation, involving dendrite-intrinsic mechanisms as well as several types of network interactions on the basis of highly selective, likely genet­ically predetermined synaptic connectivi­ty. Moreover, DS ganglion cell types appear to be more diverse than previously thought, differing not only in their preferred direction and response polarity, but also in physiology, DS mechanism, dendritic morphology and, importantly, the target area of their projec­tions in the brain.