CD15 immunoreactive amacrine cells in the mouse retina

Abstract

The mouse retina has become an important model in vision research, mainly because of the wide availability of transgenic animals. In order to study cell function and connectivity in the inner retina, antibodies that differentially stain one cell type, or a small number of cell types, are helpful as markers. Here we characterize the CD15 (3[alpha1-3]-fucosyl-N-acetyl-lactosamine)-positive cells in the mouse retina using immunofluorescence confocal microscopy and reverse-transcription polymerase chain reaction. CD15 immunoreactivity was observed in two distinct types of amacrine cells and, faintly, in some cone bipolar cells. Type I CD15+ amacrine cells are GABAergic wide-field cells that stratify in lamina 3 and 4/5 of the inner plexiform layer. Type II CD15+ amacrine cells are also GABAergic and costratify with the dopaminergic tyrosine hydroxylase-positive cells in lamina 1 of the inner plexiform layer. The densities of types I and II CD15+ amacrine cells in mid-periphery were 258 cells/mm(2) and 274 cells/mm(2). Double labeling with several other markers for amacrine cell types showed that neither type belongs to another previously identified subpopulation of amacrine cells. Single-cell RT-PCR showed that CD15+ amacrine cells coexpress several AMPA receptors - GluR1, GluR2, and GluR4 being the most common combination.

Expression of the homeodomain transcription factor Meis2 in the embryonic and postnatal retina

“…Sections were incubated with the primary antibodies overnight at 4°C, washed in PBS, and incubated for 2 hours at room temperature in Alexa594-or Alexa488-conjugated secondary antibodies (all from Molecular Probes, Eugene, OR), diluted 1:1,000 in 5% Chemiblocker and 0.5% Triton X-100. All antibodies employed in this study have been used repeatedly on paraformaldehydefixed retinal tissue before and gave similar staining patterns as described here (e.g., Mullen et al, 1992;Kawakami et al, 1997;Gabriel and Witkovsky, 1998;Dyer and Cepko, 2000;Haverkamp and Wä ssle, 2000;Toresson et al, 2000;Marquardt et al, 2001;William et al, 2001;Cuenca et al, 2003;Jakobs et al, 2003;de Melo et al, 2003;Farajian et al, 2004;Li et al, 2004;Heinze et al, 2007).…”

“…N2280). The staining pattern of the antibody against the carbohydrate epitope CD15 in the mouse retina has been established in previous studies (Jakobs et al, 2003). The specificity of the monoclonal anti-NeuN antibody has been confirmed in Western blots from adult brain cell nuclei and by immunohistochemical labeling of sec- Western blot analysis of nuclear (n) and cytoplasmic (c) extracts from adult mouse retina to assess the quality of the antibodies used.…”

“…Many GABAergic AM cells also contain other neurotransmitters like acetylcholine or dopamine. To date, only few additional molecular markers that identify distinct AM cell subpopulations have been described and these include parvalbumin, calbindin, calretinin, neural nitric oxide synthase, the cyclin kinase inhibitor p57 KIP2 , or the glycoconjugate CD15 (3[␣1-3]-fucosyl-N-acetyl-lactosamine) (Pasteels et al, 1990;Uesugi et al, 1992;Wä ssle et al, 1993;Wadhwa, 1997, 1999;Haverkamp and Wä ssle, 2000;Dyer and Cepko, 2001;Jakobs et al, 2003).…”

Cone Contacts, Mosaics, and Territories of Bipolar Cells in the Mouse Retina

“…Recoverin immunolabeling, for instance, is expressed in OFF midget bipolar cells of the macaque monkey retina (Milam et al, 1993; Wässle et al, 1994), in OFF and ON bipolar cells of the rat retina (Euler and Wässle, 1995), and in OFF bipolar cells of the mouse retina (Haverkamp et al, 2003). In the marmoset monkey, CD15 is expressed in OFF midget and DB6 ON cone bipolar cells (Chan et al, 2001), in the rabbit, CD15 labels an ON cone bipolar cell (Brown and Masland, 1999), whereas in the mouse, CD15 is expressed in OFF cone bipolar cells (Jakobs et al, 2003;Ivanova et al, 2006). These examples show that a comparison of bipolar cell types between different species has to rely on multiple features, such as the expression of glutamate receptors (kainate or AMPA) (DeVries, 2000), the axonal stratification, and the cone convergence (midget or diffuse), and at present, a unifying classification scheme of mammalian bipolar cells is not yet possible.…”

Different Functional Types of Bipolar Cells Use Different Gap-Junctional Proteins

“…The methods for dissociation of the retina are based on those described previously (Jakobs et al, 2003). In brief, a piece of retina from the 357 mouse was incubated in 2 ml of HBSS containing 0.5 mg/ml papain (Worthington, Lakewood, NJ) for 20 min at 37°C.…”