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Split Retina as an Improved Flatmount Preparation for Studying Inner Nuclear Layer Neurons in Vertebrate Retina

Bipolar cells and horizontal cells of the vertebrate retina are the first neurons to process visual information after photons are detected by ...

Oregon Health and Science University

Adaptation to changes in ambient light intensity, in retinal cells and circuits, optimizes visual functions. In the retina, light-adaptation results in changes in light-sensitivity and spatiotemporal tuning of ganglion cells. Under light-adapted conditions, contrast sensitivity (CS) of ganglion cells is a bandpass function of spatial frequency; in contrast, dark-adaptation reduces CS, especially at higher spatial frequencies. In this work, we aimed to understand intrinsic neuromodulatory mechanisms that underlie retinal adaptation to changes in ambient light level. Specifically, we investigated how CS is affected by dopamine (DA), nitric oxide (NO), and modifiers of electrical coupling through gap junctions, under different conditions of adapting illumination. Using the optokinetic response as a behavioral readout of direction-selective ganglion cell activity, we characterized the spatial CS of chicks under high- and low-photopic conditions and how it was regulated by DA, NO, and gap-junction uncouplers. We observed that: (1) DA D2R-family agonists and a donor of NO increased CS tested in low-photopic illumination, as if observed in the high-photopic light; whereas (2) removing their effects using either DA antagonists or NO- synthase inhibitors mimicked low-photopic CS; (3) simulation of high-photopic CS by DA agonists was abolished by NO-synthase inhibitors; and (4) selectively blocking coupling via connexin 35/36-containing gap junctions, using a "designer" mimetic peptide, increased CS, as does strong illumination. We conclude that, in the chicken retina: (1) DA and NO induce changes in spatiotemporal processing, similar to those driven by increasing illumination, (2) DA possibly acts through stimulating NO synthesis, and (3) blockade of coupling via gap junctions containing connexin 35/36 also drives a change in retinal CS functions. As a noninvasive method, the optokinetic response can provide rapid, conditional, and reversible assessment of retinal functions when pharmacological reagents are injected into the vitreous humor. Finally, the chick's large eyes, and the many similarities between their adaptational circuit functions and those in mammals such as the mouse, make them a promising model for future retinal research.

Light adaptation in the chick retina: Dopamine, nitric oxide, and gap-junction coupling modulate spatiotemporal contrast sensitivity.

Rod bipolar cells (RBCs) faithfully transmit light-driven signals from rod photoreceptors in the outer retina to third order neurons in the inner retina. Recently, significant work has focused on the role of leucine-rich repeat (LRR) proteins in synaptic development and signal transduction at RBC synapses. We previously identified trophoblast glycoprotein (TPBG) as a novel transmembrane LRR protein localized to the dendrites and axon terminals of RBCs.

Qing Shi

Department of Chemical Physiology and Biochemistry

Oregon Health and Science University

Split Retina as an Improved Flatmount Preparation for Studying Inner Nuclear Layer Neurons in Vertebrate Retina

Qing Shi

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Chemical Physiology and Biochemistry

Oregon Health and Science University

Split Retina as an Improved Flatmount Preparation for Studying Inner Nuclear Layer Neurons in Vertebrate Retina

Department of Chemical Physiology and Biochemistry