Abstract

<p><b>A–E</b>: magnifications from flat mounted retinas showing RGCs detected by FG tracing (<b>A</b>), Brn3a (<b>B</b>), Brn3b (<b>C</b>), Brn3c (<b>D</b>) and Brn3a+b+c immunodetection. <b>A, B</b> and <b>C, D</b> are images taken from the same retinal frame. In <b>E</b> the three Brn3 members were detected using the same fluorophore (Brn3<sup>+</sup>RGCs). <b>F–Y</b>: Representative isodensity maps showing the retinal distribution of RGCs in SD (albino) and PVG (pigmented rats). <b>F–I:</b> fluorogold traced RGCs, <b>J–M</b>: Brn3a<sup>+</sup>RGCs, <b>N–Q</b>: Brn3b<sup>+</sup>RGCs, <b>R–U</b>: Brn3c<sup>+</sup>RGCs, <b>V–Y:</b> Brn3<sup>+</sup>RGCs. For each marker and strain is shown the distribution of RGCs in one left and one right retina. Notice that, because FG and Brn3a or Brn3b and Brn3c were double detected, maps F&J, G&K, H&L, I&M, N&R, O&S, P&T and Q&U come from the same retinas. Isodensity maps are created from the data gathered after automated quantification. At the bottom of each one is shown the number of RGCs counted in the retina wherefrom the map has been generated. These maps express the RGC density according to a colour scale (bottom right in <b>I, M, Q, U</b> and <b>Y</b>) that ranges from 0 RGCs/mm<sup>2</sup> (blue) to a maximum density (red) that is 3,200 RGCs/mm<sup>2</sup> or more for FG<sup>+</sup>, Brn3a<sup>+</sup>and Brn3<sup>+</sup>RGCs; 1,800 RGCs/mm<sup>2</sup> or more for Brn3b<sup>+</sup>RGCs, and 1,600 RGCs/mm<sup>2</sup> or more for Brn3c<sup>+</sup>RGCs. The maximum density was adjusted to these numbers to allow the visualization of high and low density areas within the retina. Retinal orientation is shown in F–I: superior (S), nasal (N) temporal (T) and inferior (I). <i>Bars:</i> 20 µm (A), 1 mm (F, H).</p

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