![]() ![]() Microglia localize to the inner retina, but can migrate to the outer retina upon activation by photoreceptor damage or stress. The ganglion cell bodies populate the ganglion cell layer (GCL) together with some amacrine cells and astrocytes. ![]() Neurotransmission from second-order neurons in the INL to retinal ganglion cells occurs in the inner plexiform layer (IPL). The inner nuclear layer (INL) contains cell bodies of bipolar, horizontal, amacrine, and Müller glial cells. In the OPL, photoreceptors connect to the second-order neurons of the inner retina, which itself is morphologically and functionally organized in several layers 2. ![]() Photoreceptors are highly polarized cells consisting of a light-sensitive outer segment (OS), an inner segment (IS) containing the metabolic machinery of the cell, a soma with the nucleus in the outer nuclear layer (ONL), and an axon ending in a synaptic terminal in the outer plexiform layer (OPL). The outermost cell layer is populated by rod and cone photoreceptors. The retina is a highly heterogeneous tissue with seven major cell types arranged in three cellular layers connected by two synaptic layers, providing visual input detection and initial signal processing 1 (Fig. ReLayS is a simple and useful method to address protein and possibly metabolites distribution in photoreceptor compartments in various situations including development, ageing, and degenerative diseases. Furthermore, the separation of the PS from the ONL enabled the detection of light-driven translocation of transducin from the PS to the soma. The separation of PS, ONL, and InR was successfully validated by Western blotting and real-time PCR using proteins and genes with known expression profiles within the retina. The layer-specific material isolated from a mouse half-retina with the ReLayS method was sufficient for protein isolation and Western blotting or RNA isolation and real-time PCR studies. Here we present the ReLayS method-a simple technique for the separation of photoreceptor segments (PS) containing both inner and outer segments, outer nuclear layer (ONL), and inner retina (InR) that contains the remaining retinal layers. However, the molecular analysis of photoreceptors is a challenge due to the heterogeneity of the retinal tissue and the lack of easy and reliable methods for cell separation. Understanding the physiology of the retina, and especially of the highly polarized photoreceptors, is essential not only to broaden our knowledge of the processes required for normal vision, but also to develop effective therapies to prevent or slow retinal degenerative diseases. ![]()
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