Immunocytochemical staining of AII‐amacrine cells in the rat retina with antibodies against parvalbumin

Abstract

The rod dominated rodent retina is the preferred tissue for in vitro studies of mammalian retinal physiology and pharmacology. The rod pathway through the rat retina was investigated, therefore, in order to find out whether its organization follows the mammalian "plan." AII-amacrine cells of the rat retina were injected with Lucifer Yellow to characterize the morphology of this bistratified interneuron of the rod pathway. When sections or whole mounts of the rat retina were stained with antibodies against the calcium binding protein parvalbumin (PV), two different amacrine cell types were labeled: the AII-amacrine cell and a widefield amacrine cell. They occur at a ratio of 12:1. Weak label was also observed in ganglion cells. The density of PV-labeled AII-cells decreases from approximately 7,000 cells/mm2 in upper central retina to 2,000 cells/mm2 in peripheral retina. Their cell bodies form a regular mosaic, and the dendritic arbors of three neighbouring AII-amacrine cells overlap (coverage of 3).

Electrical synapses between AII amacrine cells in the retina: Function and modulation

“…One specific type of amacrine input targets the cell body region and originates from dopaminergic cells (Kolb et al, 1990;Voigt and Wässle, 1987) which might use GABA (Wässle and Chun, 1988) and/or glutamate (Anderson et al, 2011) as a co-transmitter. Processes from the dopaminergic amacrine cells form ring-like structures around the cell bodies / apical dendrites of AII amacrines and ultrastructural examination has demonstrated the presence of specialized synaptic contacts (Kolb et al, 1990;Voigt and Wässle, 1987;Wässle et al, 1993). This innervation is of particular interest because of the evidence for dopaminergic modulation of tracer coupling between AII amacrine cells (Hampson et al, 1992; see below).…”

“…Specificity of signaling can be achieved and maintained via Ca 2+ nano-or microdomains, i.e., the tight physical coupling between a Ca 2+ source and a downstream signaling pathway. Whereas there are at least three possible sources for Ca 2+ influx in AII amacrines (CP-GluARs, GluNRs and voltagegated Ca 2+ channels), the expression of mobile, cytoplasmic Ca 2+ buffers (parvalbumin, calretinin, calbindin) in these cells (Casini et al, 1995;Goebel and Pourcho, 1997;Massey and Mills, 1999a;Wässle et al, 1993Wässle et al, , 1995 could be important mechanisms for spatially localizing Ca 2+ signals. In addition to Ca 2+ influx and buffering, Ca 2+ dynamics are also influenced by Ca 2+ clearance, with two acknowledged mechanisms in dendrites: uptake of Ca 2+ into intracellular stores via (sarco)endoplasmic reticulum Ca 2+ -ATPase (SERCA) pumps and extrusion of Ca 2+ through the plasma membrane via a Ca 2+ -ATPase and a Na + /Ca 2+ exchanger (Clapham, 2007;Goldberg and Yuste, 2005 …”

Cellular tropism and transduction properties of seven adeno-associated viral vector serotypes in adult retina after intravitreal injection

“…Selected slides were stained with a primary rabbit or mouse anti-GFP antibody (1:100; Millipore, Sydney, Australia), followed by an appropriate fluorescein isothiocyanate-conjugated secondary antibody (1:100; Sigma), to assure that a weak native GFP fluorescence was not overlooked in the analysis. Antibodies used for GFP + cell characterization were mouse anti-bIII-tubulin (RGCs; 9,66 74 1:5000; Swant, Bellinzona, Switzerland), overnight at +4 1C, followed by 3 Â 5 min washes in PBS. Sections were then incubated for 1 h with an anti-mouse or anti-rabbit Cy3-conjugated secondary antibody (1:300; Jackson, West Grove, PA, USA), rinsed for 3 Â 5 min in PBS before being coverslipped with Dako fluorescent mounting media containing Hoechst 33342 (Sigma) nuclear dye.…”

Natural sleep modifies the rat electroretinogram.

“…The rodent retina is dominated by rods, and the connections these make within the retina are like those found in other mammals (13,14). Its two types of cones, which make up <1% of the photoreceptors, are green and blue sensitive (15), which suggests that this retina responds to shortwave and middleto-longwave light through cones and to red light through rods.…”