Identification of AⅡ amacrine, displaced amacrine, and bistratified ganglion cell types in human retina with antibodies against calretinin

Abstract

Antibodies against calretinin are markers for one type of rod pathway interneuron (AⅡ amacrine cell) in the retina of some but not all mammalian species. The AⅡ cells play a crucial role in night-time (scotopic) vision and have been proposed as a target for optogenetic restoration of vision in retinal disease. In the present study we aimed to characterize the AⅡ cells in human retina. Postmortem human donor eyes were obtained with ethical approval and processed for calretinin immunofluorescence. Calretinin-positive somas in the inner nuclear and the ganglion cell layer were filled with the lipophilic dye DiI. The large majority (over 80%) of calretinin-immunoreactive cells is located in the inner nuclear layer, is immunopositive for glycine transporter 1, and shows the typical morphology of AⅡ amacrine cells. In addition, a small proportion of calretinin-positive cells in the inner nuclear layer and in the ganglion cell layer is glutamic acid decarboxylase-positive and shows the morphology of widefield amacrine cells (stellate, semilunar, and thorny amacrine cells). About half of the calretinin cells in the ganglion cell layer are bistratified ganglion cells resembling the small bistratified (presumed blue-ON/yellow-OFF) and the G17 ganglion cell previously described in primates. We conclude that in human retina, antibodies against calretinin can be used to identify AⅡ amacrine cells in the inner nuclear layer as well as widefield amacrine and small bistratified ganglion cells in the ganglion cell layer.

Connexin36 Expression in the Mammalian Retina: A Multiple-Species Comparison

“…CaR has been demonstrated to specifically label AII ACs in most mammalian species (rabbit – Völgyi et al, 1997; Massey and Mills, 1999; monkey – Mills and Massey, 1999; cat – Goebel and Pourcho, 1997; human – Kántor et al, 2016b; Lee et al, 2016), whereas starburst but not AII ACs express CaR in the rat and the mouse retina (Gábriel and Witkovsky, 1998; Haverkamp and Wässle, 2000). These observations were confirmed here showing that AII ACs in the rabbit, cat, and monkey retina express CaR, whereas AII ACs in the rat and the mouse did not.…”

Prox1 Is a Marker for AII Amacrine Cells in the Mouse Retina

“…GABA-immunoreactive amacrine cells are characterized by medium and wide-field processes, and several wide-field types contain a second neuroactive substance, including vasoactive intestinal polypeptide, substance P, acetylcholine, or dopamine (Brecha et al, 1988; Wässle and Chun, 1988; Vaney et al, 1989; Casini and Brecha, 1992; Strettoi and Masland, 1996; Akrouh and Kerschensteiner, 2015; Park et al, 2015). In contrast, the glycine-immunoreactive amacrine cells have narrow-field processes that span multiple IPL laminae, and several types also contain a second immunohistochemical marker, including parvalbumin, calretinin and Disabled 1 (Wässle et al, 1993; Haverkamp and Wässle, 2000, 2004; Rice and Curran, 2000; Lee et al, 2006, 2016). An exception to this general principle is that VGluT3-immunoreactive amacrine cells, which have medium-field processes distributed to multiple IPL laminae, exhibit both glycine and glutamate immunoreactivity (Haverkamp and Wässle, 2004; Johnson et al, 2004; Grimes et al, 2011; Kim et al, 2015).…”

“…Another approach identifying AII amacrine cells is using immunohistochemistry with antibodies to parvalbumin in rabbit (Casini et al, 1995) and rat (Wässle et al, 1993), while antibodies to calretinin have been used in cat (Pasteels et al, 1990; Gábriel and Straznicky, 1992; Macneil et al, 2009), macaque (Wässle et al, 1995; Massey and Mills, 1999; Kolb et al, 2002), rabbit (Massey and Mills, 1999) and human (Lee et al, 2004, 2016). Lastly, disabled 1 has been used in mouse (Rice and Curran, 2000; Lee et al, 2004, 2006).…”

Thorny ganglion cells in marmoset retina: Morphological and neurochemical characterization with antibodies against calretinin

“…In primate retina, the expression of calcium binding proteins shows common pattern as well as species specific differences (Chiquet, Dkhissi‐Benyahya, & Cooper, ; Pasteels, Rogers, Blachier, & Pochet, ). For example, in all primate retinas studied to date, calretinin is expressed in AII amacrine cells (macaque: Kolb, Zhang, Dekorver, & Cuenca, ; Mills & Massey, ; Wässle, Grünert, Chun, & Boycott, , marmoset: Jusuf, Lee, & Grünert, ; Weltzien, Percival, Martin, & Grünert, , and human: Lee, Weltzien, Madigan, Martin, & Grünert, ). The expression of calretinin in the ganglion cell layer, however, varies between primate species.…”

“…In macaque retina, calretinin expression in the ganglion cell layer is downregulated after birth (Hendrickson, Yan, Erickson, Possin, & Pow, ), thus these cells are rare in the adult. In human retina, the majority of calretinin expressing cells in the ganglion cell layer are displaced amacrine cells, and calretinin expressing ganglion cells have been identified as bistratified cells (Lee et al, ). In marmoset, calretinin expressing cells make up a clear subpopulation of cells in the ganglion cell layer (Chiquet et al, ; Jusuf et al, ) but the cell types have not been identified.…”