Enhancing the dark side: asymmetric gain of cone photoreceptors underpins their discrimination of visual scenes based on skewness.

Psychophysical data indicate that humans can discriminate visual scenes based on their skewness, i.e. the ratio of dark and bright patches within a visual scene. It has also been shown that at a phenomenological level this skew discrimination is described by the so-called blackshot mechanism, which accentuates strong negative contrasts within a scene. Here, we present a set of observations suggesting that the underlying computation might start as early as the cone phototransduction cascade, whose gain is higher for strong negative contrasts than for strong positive contrasts. We recorded from goldfish cone photoreceptors and found that the asymmetry in the phototransduction gain leads to responses with larger amplitudes when using negatively rather than positively skewed light stimuli. This asymmetry in amplitude was present in the cone photocurrent, voltage response and synaptic output. Given that the properties of the phototransduction cascade are universal across vertebrates, it is possible that the mechanism shown here gives rise to a general ability to discriminate between scenes based only on their skewness, which psychophysical studies have shown humans can do. Thus, our data suggest the importance of non-linearity of the early photoreceptor for perception. Additionally, we found that stimulus skewness leads to a subtle change in photoreceptor kinetics. For negatively skewed stimuli, the impulse response functions of the cone peak later than for positively skewed stimuli. However, stimulus skewness does not affect the overall integration time of the cone. KEY POINTS: Humans can discriminate visual scenes based on skewness, i.e. the relative prevalence of bright and dark patches within a scene. Here, we show that negatively skewed time-series stimuli induce larger responses in goldfish cone photoreceptors than comparable positively skewed stimuli. This response asymmetry originates from within the phototransduction cascade, where gain is higher for strong negative contrasts (dark patches) than for strong positive contrasts (bright patches). Unlike the implicit assumption often contained within models of downstream visual neurons, our data show that cone photoreceptors do not simply relay linearly filtered versions of visual stimuli to downstream circuitry, but that they also emphasize specific stimulus features. Given that the phototransduction cascade properties among vertebrate retinas are mostly universal, our data imply that the skew discrimination by human subjects reported in psychophysical studies might stem from the asymmetric gain function of the phototransduction cascade.

High Contrast Allows the Retina to Compute More Than Just Contrast.

The goal of sensory processing is to represent the environment of an animal. All sensory systems share a similar constraint: they need to encode a wide range of stimulus magnitudes within their narrow neuronal response range. The most efficient way, exploited by even the simplest nervous systems, is to encode relative changes in stimulus magnitude rather than the absolute magnitudes. For instance, the retina encodes contrast, which are the variations of light intensity occurring in time and in space. From this perspective, it is easy to understand why the bright plumage of a moving bird gains a lot of attention, while an octopus remains motionless and mimics its surroundings for concealment. Stronger contrasts simply cause stronger visual signals. However, the gains in retinal performance associated with higher contrast are far more than what can be attributed to just a trivial linear increase in signal strength. Here we discuss how this improvement in performance is reflected throughout different parts of the neural circuitry, within its neural code and how high contrast activates many non-linear mechanisms to unlock several sophisticated retinal computations that are virtually impossible in low contrast conditions.

Measurement of intracellular concentration of fluorescently-labeled targets in living cells.

Estimations of intracellular concentrations of fluorescently-labeled molecules within living cells are very important for guidance of biological experiments and interpretation of their results. Here we propose a simple and universal approach for such estimations. The approach is based upon common knowledge that the dye fluorescence is directly proportional to its quantum yield and the number of its molecules and that a coefficient of proportionality is determined by spectral properties of the dye and optical equipment used to record fluorescent signals. If two fluorescent dyes are present in the same volume, then a ratio of their concentrations is equal to a ratio of their fluorescence multiplied by some dye- and equipment-dependent coefficient. Thus, if the coefficient and concentration of one dye is known then the concentration of another dye can be determined. Here we have demonstrated how to calculate this coefficient (called a ratio factor) and how to use it for concentration measurements of fluorescently tagged molecules. As an example of how this approach can be used, we estimated a concentration of exogenously expressed neuronal Ca2+ sensor protein, hippocalcin, tagged by a fluorescent protein in a dendritic tree of rat hippocampal neurons loaded via a patch pipette with Alexa Fluor dye of known concentration. The new approach should allow performing a fast, inexpensive and reliable quantitative analysis of fluorescently-labeled targets in different parts of living cells.