Intrinsic optical signal (IOS) imaging promises to be a noninvasive method for high-resolution examination of retinal physiology, which can advance the study and diagnosis of eye diseases. While specialized optical instruments are desirable for functional IOS imaging of retinal physiology, in depth understanding of multiple IOS sources in the complex retinal neural network is essential for optimizing instrument designs. We provide a brief overview of IOS studies and relationships in rod outer segment suspensions, isolated retinas, and intact eyes. Recent developments of line-scan confocal and functional optical coherence tomography (OCT) instruments have allowed in vivo IOS mapping of photoreceptor physiology. Further improvements of the line-scan confocal and functional OCT systems may provide a feasible solution to pursue functional IOS mapping of human photoreceptors. Some interesting IOSs have already been detected in inner retinal layers, but better development of the IOS instruments and software algorithms is required to achieve optimal physiological assessment of inner retinal neurons.

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Intrinsic optical signal imaging of retinal physiology: a review.

Stimulus-evoked intrinsic optical signal (IOS), which occurs almost immediately after the onset of retinal stimulus has been observed in retinal photoreceptors, promises to be a unique biomarker for objective optoretinography (ORG) of photoreceptor function. We report here the first-time in vivo ORG detection of photoreceptor dysfunction due to retinal degeneration. A custom-designed optical coherence tomography (OCT) was employed for longitudinal ORG monitoring of photoreceptor-IOS distortions in retinal degeneration mice. Depth-resolved OCT analysis confirmed the outer segment (OS) as the physical source of the photoreceptor-IOS. Comparative ERG measurement verified the phototransduction activation as the physiological correlator of the photoreceptor-IOS. Histological examination revealed disorganized OS discs, i.e. the pathological origin of the photoreceptor-IOS distortion.

Functional optical coherence tomography enables in vivo optoretinography of photoreceptor dysfunction due to retinal degeneration.

Transient retinal phototropism (TRP) has been predominantly observed in rod photoreceptors activated by oblique visible light stimulation. Dynamic confocal microscopy and optical coherence tomography (OCT) have revealed rod outer segment (ROS) movement as the physical source of TRP. However, the physiological source of ROS movement is still not well understood. In this study, concurrent near-infrared imaging of TRP and electroretinogram (ERG) measurement of retinal electrophysiology revealed that ROS movement occurs before the onset of the ERG a-wave, which is known to reflect the hyperpolarization of retinal photoreceptors. Moreover, substitution of normal superfusing medium with low-sodium medium reversibly blocked the photoreceptor ERG a-wave, but largely preserved the stimulus-evoked ROS movements. Our experimental results and theoretical analysis indicate that early, disc-based stages of the phototransduction cascade, which occur before the hyperpolarization of retinal photoreceptors, contribute to the TRP associated ROS movement.

Stimulus-evoked outer segment changes occur before the hyperpolarization of retinal photoreceptors.

A recent development of kinetic light-scattering and -absorption photometry is described. Essential points are:
 The method allows comparative kinetic analysis of absorption and scattering signals. The possible causal connections between pigment and membrane structure processes can be selected.

Comparative kinetic light-scattering and -absorption photometry

Purpose This study was designed to verify light-induced outer segment (OS) length shrinkage of rod photoreceptors and to characterize its anatomic source at disc-level resolution. Methods Frog (Rana pipiens) retinas were used for this study. Time-lapse light microscopy of freshly isolated OSs was employed to test transient rod OS changes at 10 ms temporal resolution. Histological light microscopy of dark- and light-adapted retinas was used to confirm light-induced rod OS length changes; and transmission electron microscopy (TEM) was used to quantify light-driven structural perturbation of rod OSs at disc level resolution. Results Time-lapse light microscopy images verified transient length shrinking responses in freshly isolated rod OSs. Histological light microscopy images confirmed reduced rod OS lengths in light-adapted retinas, compared to that of dark-adapted retinas. TEM images disclosed shortened inter-disc distances in light-adapted retinas compared to dark-adapted retinas. Conclusions Light-induced rod OS length shrinkage was confirmed using time-lapse light microscopy of isolated rod OSs and histological light microscopy of dark- and light-adapted retinas. TEM revealed that the rod OS length shrinkage was correlated to the light-driven decrease of the space between individual discs, not the disc thickness itself. Translational relevance Light-induced transient rod response promises a noninvasive biomarker for early diagnosis of age-related macular degeneration and retinitis pigmentosa, in which the rod photoreceptors are known to be more vulnerable than cone photoreceptors.

H. Asai

Papers

A light-induced conformational change in rod photoreceptor disc membrane.