DNER and NFIA are expressed by developing and mature AII amacrine cells in the mouse retina

Abstract

The present study has taken advantage of publicly available cell type specific mRNA expression databases in order to identify potential genes participating in the development of retinal AII amacrine cells. We profile two such genes, Delta/Notch-like EGF repeat containing (Dner) and nuclear factor I/A (Nfia), that are each heavily expressed in AII amacrine cells in the mature mouse retina, and which conjointly identify this retinal cell population in its entirety when using antibodies to DNER and NFIA. DNER is present on the plasma membrane, while NFIA is confined to the nucleus, consistent with known functions of each of these two proteins. DNER also identifies some other subsets of retinal ganglion and amacrine cell types, along with horizontal cells, while NFIA identifies a subset of bipolar cells as well as Muller glia and astrocytes. During early postnatal development, NFIA labels astrocytes on the day of birth, AII amacrine cells at postnatal (P) day 5, and Muller glia by P10, when horizontal cells also transiently exhibit NFIA immunofluorescence. DNER, by contrast, is present in ganglion and amacrine cells on P1, also labeling the horizontal cells by P10. Developing AII amacrine cells exhibit accumulating DNER labeling at the dendritic stalk, labeling that becomes progressively conspicuous by P10, as it is in maturity. This developmental time course is consistent with a prospective role for each gene in the differentiation of AII amacrine cells.

The somal patterning of the AII amacrine cell mosaic in the mouse retina is indistinguishable from random simulations matched for density and constrained by soma size

“…In addition, for each sampled field we generated a random simulation of cells matched in density, constraining their positioning in the simulation by the size of the AII amacrine cell somata, having previously been determined from a sample of 200 AII amacrine cells to be 7.5 ± 0.5 µ m in diameter, using antibodies that reveal the somal membrane (Keeley & Reese, 2018). The simulations randomly draw soma sizes from a Gaussian distribution with the same mean and standard deviation, where each randomly positioned cell is rejected (with replacement) if it overlaps that of a previously positioned cell, until the number of cells in the simulation matches that for the real field being simulated.…”

A single-cell guide to retinal development: Cell fate decisions of multipotent retinal progenitors in scRNA-seq

“…These include examinations of the Nfi transcription factors and the somatostatin receptor Sstr2 , that exhibit enriched expression within late RPCs and neurogenic RPCs, respectively ( Clark et al, 2019 ). During retinal development, the Nfi transcription factors Nfia, Nfib, and Nfix are each enriched within primary RPCs during late periods of retinal development ( Clark et al, 2019 ) and display restricted expression patterns within mature retinal cell types including amacrine cells ( Keeley and Reese, 2018 ; Yan et al, 2020a ), bipolar cells ( Shekhar et al, 2016 ), and Müller glia ( Clark et al, 2019 ; de Melo et al, 2016 ). Conditional knockout of Nfia/b/x within the developing retina results in a failure of late RPCs to exit the cell-cycle and differentiate as bipolar cells or Müller glia.…”

Molecular identification of sixty-three amacrine cell types completes a mouse retinal cell atlas

“…These and other DE genes allowed assignment of several clusters to previously identified AC types (Table 1). They included starburst ACs (C17: Chat; Vaney et al, 2012), Aii ACs (C3: Gjd2, Prox1, Dab1, Nfia, Dner; Hansen et al, 2005;Rice and Curran, 2000;Perez de Sevilla Müller et al, 2017;Keeley and Reese, 2018), SEGs (C4: Satb2, Ebf3, and Glyt1 [Slc6a9]; Kay et. al, 2011), VG3 ACs (C13: VGlut3 [Slc17a8]; Haverkamp and Wässle, 2004;Johnson et al, 2004;Krishnaswamy et al, 2015); and A17 ACs (C6: Prkca (PKCα), Sdk1, Calb2 negative, Dab1-negative; Grimes et al, 2010;Puthussery and Fletcher 2007;Yamagata and Sanes, 2019).…”