Abstract

The retina decomposes visual stimuli into parallel channels that encode different features of the visual environment. Central to this computation is the synaptic processing in a dense layer of neuropil, the so-called inner plexiform layer (IPL). Here, different types of bipolar cells stratifying at distinct depths relay the excitatory feedforward drive from photoreceptors to amacrine and ganglion cells. Current experimental techniques for studying processing in the IPL do not allow imaging the entire IPL simultaneously in the intact tissue. Here, we extend a two-photon microscope with an electrically tunable lens allowing us to obtain optical vertical slices of the IPL, which provide a complete picture of the response diversity of bipolar cells at a “single glance”. The nature of these axial recordings additionally allowed us to isolate and investigate batch effects, i.e. inter-experimental variations resulting in systematic differences in response speed. As a proof of principle, we developed a simple model that disentangles biological from experimental causes of variability and allowed us to recover the characteristic gradient of response speeds across the IPL with higher precision than before. Our new framework will make it possible to study the computations performed in the central synaptic layer of the retina more efficiently.

Highlights

  • The retina decomposes visual stimuli into parallel channels that encode different features of the visual environment

  • The latter is critical if UV stimuli are used: Since prolonged exposure to UV light can degrade the optomechanical properties of the lens, electrically tunable lens (ETL) are typically equipped with a UV-reflecting glass window

  • We found that batch effects alone accounted for a larger fraction of the variance than inner plexiform layer (IPL) depth (Fig. 5B), which suggests that accounting for such variation can greatly facilitate any analysis of functional differences between bipolar cells (BCs) types beyond On vs. Off

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Summary

Introduction

The retina decomposes visual stimuli into parallel channels that encode different features of the visual environment. We extend a two-photon microscope with an electrically tunable lens allowing us to obtain optical vertical slices of the IPL, which provide a complete picture of the response diversity of bipolar cells at a “single glance”. Each BC type constitutes a particular feature channel, with certain temporal dynamics[7], including On and Off BC types sensitive to light increments or decrements, respectively[14], different kinetics[15,16], and chromatic signals[17,18] Some of these features are systematically mapped across the IPL: For example, On BCs project to the inner and Off BCs to the outer portion of the IPL14,19. Wavemetrics, Lake Oswego, OR Written by T.E, supported by Tom Boissonnet (EMBL, Monterotondo)

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