Dendritic and axonal targeting patterns of a genetically-specified class of retinal ganglion cells that participate in image-forming circuits.

BackgroundThere are numerous functional types of retinal ganglion cells (RGCs), each participating in circuits that encode a specific aspect of the visual scene. This functional specificity is derived from distinct RGC morphologies and selective synapse formation with other retinal cell types; yet, how these properties are established during development remains unclear. Islet2 (Isl2) is a LIM-homeodomain transcription factor expressed in the developing retina, including approximately 40% of all RGCs, and has previously been implicated in the subtype specification of spinal motor neurons. Based on this, we hypothesized that Isl2+ RGCs represent a related subset that share a common function.ResultsWe morphologically and molecularly characterized Isl2+ RGCs using a transgenic mouse line that expresses GFP in the cell bodies, dendrites and axons of Isl2+ cells (Isl2-GFP). Isl2-GFP RGCs have distinct morphologies and dendritic stratification patterns within the inner plexiform layer and project to selective visual nuclei. Targeted filling of individual cells reveals that the majority of Isl2-GFP RGCs have dendrites that are monostratified in layer S3 of the IPL, suggesting they are not ON-OFF direction-selective ganglion cells. Molecular analysis shows that most alpha-RGCs, indicated by expression of SMI-32, are also Isl2-GFP RGCs. Isl2-GFP RGCs project to most retino-recipient nuclei during early development, but specifically innervate the dorsal lateral geniculate nucleus and superior colliculus (SC) at eye opening. Finally, we show that the segregation of Isl2+ and Isl2- RGC axons in the SC leads to the segregation of functional RGC types.ConclusionsTaken together, these data suggest that Isl2+ RGCs comprise a distinct class and support a role for Isl2 as an important component of a transcription factor code specifying functional visual circuits. Furthermore, this study describes a novel genetically-labeled mouse line that will be a valuable resource in future investigations of the molecular mechanisms of visual circuit formation

The Globular Cluster System in the Inner Region of the Giant Elliptical Galaxy NGC 4472

We present a study of globular clusters in the inner region of the giant elliptical galaxy NGC 4472, based on the HST WFPC2 archive data. We have found about 1560 globular cluster candidates at the galactocentric radius r < 4 arcmin. V-(V-I) diagram of these objects shows a dominant vertical structure which consists obviously of two components: blue globular clusters (BGCs) and red globular clusters (RGCs). The luminosity function of the globular clusters is derived to have a peak at V(max)=23.50+/-0.16 from Gaussian fitting. The distance to NGC 4472 is estimated to be d=14.7+/-1.3 Mpc.The peak luminosity for the RGCs is similar to that for the BGCs, which indicates that the RGCs may be several Gyrs younger than the BGCs. The mean luminosity of the bright BGCs decreases by 0.2 mag with increasing galactocentric radius over the range of 9 arcmin, while that of the RGCs does not. The observed color distribution of these globular clusters is distinctively bimodal with peaks at (V-I) = 0.98 and 1.23. The mean observed color of all the globular clusters with V < 23.9 mag is derived to be (V-I)=1.11. These colors are exactly the same as those for the globular clusters in M87. It is found that the relative number of the BGCs to the RGCs is increasing with the increasing galactocentric radius. Surface number density profiles of both the BGCs and RGCs get flat in the central region, and the core radii of the globular cluster systems are measured to be r_c = 1.9 arcmin for the BGCs, r_c = 1.2 arcmin for the RGCs, and r_c = 1.3 arcmin for the total sample, which are much larger than the stellar core of the galaxy. In general the properties of the globular clusters in the inner region of NGC 4472 are consistent with those of the globular clusters in the outer region of NGC 4472.Comment: 27 pages (AASLaTex), 22 Postscript Figures, Accepted for Publication in the Astronomical Journal, Jul. 31st, 200

Investigating the Role of FGF8 Signaling in Neurogenesis of the Developing Zebrafish Eye

In the embryonic zebrafish, the fibroblast growth factor 8a (FGF8) signaling network is essential for proper development and maintenance of retinal ganglion cells (RGCs) as well as the hyaloid vasculature, the vessels that supply the eye with nutrients during development. Disruption of FGF8 signaling via knock down of FGF8 or pharmacologic inhibition of FGF receptors (FGFRs) results in extensive abnormalities throughout the developing eye. Our preliminary data indicated that in developing zebrafish, mRNA expression of fgf8a is present exclusively in the RGCs, while the fibroblast growth factor receptor 1 (fgfr1b) is expressed exclusively in the area of the hyaloid vasculature. These results led us to hypothesize that FGF8 signals from the RGCs to the vasculature of the developing eye, and that this signaling network is essential for proper eye development. In order to test this hypothesis, we demonstrated the ability to detect downstream phosphorylation events in response to acute FGF8 stimulation in cells that expressed FGFR1 using Western blot and immunofluorescence (IF). Next, we established a zebrafish eye explant culture system to treat the cells of the developing zebrafish eye in vitro. Using transgenic zebrafish lines expressing green fluorescent protein (GFP) tags in either the differentiating RGCs or the vascular cells of the eye, we attempted to identify the specific cells capable of responding to FGF8. Our data indicate that recombinant FGF8 is capable of activating detectable intracellular signaling cascades, such as ERK phosphorylation, in cultured endothelial cells. Furthermore, FGF8 is capable of inducing signaling in some of the cells from the developing zebrafish eye, but not in the RGCs. These findings support our proposed model in which FGF8 signals from the RGCs to the hyaloid vasculature, resulting in the activation of signaling pathways that are necessary for proper development of the hyaloid vasculature and RGCs

Nel positively regulates the genesis of retinal ganglion cells by promoting their differentiation and survival during development

Peer reviewedPublisher PD

Elevated expression of human bHLH factor ATOH7 accelerates cell cycle progression of progenitors and enhances production of avian retinal ganglion cells.

The production of vertebrate retinal projection neurons, retinal ganglion cells (RGCs), is regulated by cell-intrinsic determinants and cell-to-cell signaling events. The basic-helix-loop-helix (bHLH) protein Atoh7 is a key neurogenic transcription factor required for RGC development. Here, we investigate whether manipulating human ATOH7 expression among uncommitted progenitors can promote RGC fate specification and thus be used as a strategy to enhance RGC genesis. Using the chicken retina as a model, we show that cell autonomous expression of ATOH7 is sufficient to induce precocious RGC formation and expansion of the neurogenic territory. ATOH7 overexpression among neurogenic progenitors significantly enhances RGC production at the expense of reducing the progenitor pool. Furthermore, forced expression of ATOH7 leads to a minor increase of cone photoreceptors. We provide evidence that elevating ATOH7 levels accelerates cell cycle progression from S to M phase and promotes cell cycle exit. We also show that ATOH7-induced ectopic RGCs often exhibit aberrant axonal projection patterns and are correlated with increased cell death during the period of retinotectal connections. These results demonstrate the high potency of human ATOH7 in promoting early retinogenesis and specifying the RGC differentiation program, thus providing insight for manipulating RGC production from stem cell-derived retinal organoids

Inhibitory control of feature selectivity in an object motion sensitive circuit of the retina

Object motion sensitive (OMS) W3-retinal ganglion cells (W3-RGCs) in mice respond to local movements in a visual scene but remain silent during self-generated global image motion. The excitatory inputs that drive responses of W3-RGCs to local motion were recently characterized, but which inhibitory neurons suppress W3-RGCs’ responses to global motion, how these neurons encode motion information, and how their connections are organized along the excitatory circuit axis remains unknown. Here, we find that a genetically identified amacrine cell (AC) type, TH2-AC, exhibits fast responses to global motion and slow responses to local motion. Optogenetic stimulation shows that TH2-ACs provide strong GABAA receptor-mediated input to W3-RGCs but only weak input to upstream excitatory neurons. Cell-type-specific silencing reveals that temporally coded inhibition from TH2-ACs cancels W3-RGC spike responses to global but not local motion stimuli and, thus, controls the feature selectivity of OMS signals sent to the brain

AAV-mediated and pharmacological induction of Hsp70 expression stimulates survival of retinal ganglion cells following axonal injury.

We evaluated the effect of AAV2- and 17-AAG (17-N-allylamino-17-demethoxygeldanamycin)-mediated upregulation of Hsp70 expression on the survival of retinal ganglion cells (RGCs) injured by optic nerve crush (ONC). AAV2-Hsp70 expression in the retina was primarily observed in the ganglion cell layer. Approximately 75% of all transfected cells were RGCs. RGC survival in AAV2-Hsp70-injected animals was increased by an average of 110% 2 weeks after the axonal injury compared with the control. The increase in cell numbers was not even across the retinas with a maximum effect of approximately 306% observed in the inferior quadrant. 17-AAG-mediated induction of Hsp70 expression has been associated with cell protection in various models of neurodegenerative diseases. We show here that a single intravitreal injection of 17-AAG (0.2 ug ul(-1)) results in an increased survival of ONC-injured RGCs by approximately 49% compared with the vehicle-treated animals. Expression of Hsp70 in retinas of 17-AAG-treated animals was upregulated approximately by twofold compared with control animals. Our data support the idea that the upregulation of Hsp70 has a beneficial effect on the survival of injured RGCs, and the induction of this protein could be viewed as a potential neuroprotective strategy for optic neuropathies

Tbr1 instructs laminar patterning of retinal ganglion cell dendrites.

Visual information is delivered to the brain by &gt;40 types of retinal ganglion cells (RGCs). Diversity in this representation arises within the inner plexiform layer (IPL), where dendrites of each RGC type are restricted to specific sublaminae, limiting the interneuronal types that can innervate them. How such dendritic restriction arises is unclear. We show that the transcription factor Tbr1 is expressed by four mouse RGC types with dendrites in the outer IPL and is required for their laminar specification. Loss of Tbr1 results in elaboration of dendrites within the inner IPL, while misexpression in other cells retargets their neurites to the outer IPL. Two transmembrane molecules, Sorcs3 and Cdh8, act as effectors of the Tbr1-controlled lamination program. However, they are expressed in just one Tbr1+ RGC type, supporting a model in which a single transcription factor implements similar laminar choices in distinct cell types by recruiting partially non-overlapping effectors