The complex, yet highly ordered and predictable, structure of the neural retina is one of the most conserved features of the vertebrate central nervous system. In all vertebrate classes, retinal neurons are organized into laminae with each neuronal class adopting specific morphologies and patterns of connectivity. Using genetic analyses in zebrafish, we demonstrate that N-cadherin (Ncad) has several distinct and crucial functions during the establishment of retinal organization. Although the location of cell division is disorganized in embryos with reduced or no Ncad function, different classes of retinal neurons are generated. However, these neurons fail to organize into correct laminae, most probably owing to compromised adhesion between retinal cells. In addition, amacrine cells exhibit exuberant and misdirected outgrowth of neurites that contributes to severe disorganization of the inner plexiform layer. Retinal ganglion cells also exhibit defects in process outgrowth, with axons exhibiting fasciculation defects and adopting incorrect ipsilateral trajectories. At least some of these defects are likely to be due to a failure to maintain compartment boundaries between eye, optic nerve and brain. Although in vitro studies have implicated Fgf receptors in modulating the axon outgrowth promoting properties of Ncad, most aspects of the Ncad mutant phenotype are not phenocopied by treatments that block Fgf receptor function.

Glycerophospholipids, the structural components of cell membranes, have not been considered to be spatial cues for intercellular signaling because of their ubiquitous distribution. We identified lyso-phosphatidyl-β-D-glucoside (LysoPtdGlc), a hydrophilic glycerophospholipid, and demonstrated its role in modality-specific repulsive guidance of spinal cord sensory axons. LysoPtdGlc is locally synthesized and released by radial glia in a patterned spatial distribution to regulate the targeting of nociceptive but not proprioceptive central axon projections. Library screening identified the G protein-coupled receptor GPR55 as a high-affinity receptor for LysoPtdGlc, and GPR55 deletion or LysoPtdGlc loss of function in vivo caused the misallocation of nociceptive axons into proprioceptive zones. These findings show that LysoPtdGlc/GPR55 is a lipid-based signaling system in glia-neuron communication for neural development.

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The cell adhesion molecule L1 plays crucial roles in axon tract development. In vitro, L1 presented as a culture substrate stimulates axon elongation by binding to L1 expressed on the growth cone. In migrating growth cones, L1 is endocytosed via the AP-2/clathrin-mediated pathway at the central domain, followed by anterograde vesicular transport and recycling to the plasma membrane of the leading front. It has previously been shown that this endocytic trafficking of L1 is critical for axon elongation (Kamiguchi and Yoshihara [2001] J. Neurosci. 21:9194-9203). Adjacent to the AP-2 recognition site, the L1 cytoplasmic domain has a cluster of acidic amino acids containing Ser1181 that can be phosphorylated by casein kinase II (CKII; Wong et al. [1996a] J. Neurochem. 66:779-786). In this paper, we demonstrate that Ser1181 phosphorylation by CKII is implicated in both normal endocytic trafficking of L1 and L1-stimulated axon growth. Whereas L1 is sorted into transferrin-positive endosomes after endocytosis, pharmacological inhibition of CKII caused some population of L1 to be internalized into transferrin-negative compartments. Single-amino-acid mutations at Ser1181, which either prevent or mimic phosphorylation by CKII, caused similar missorting of internalized L1. Furthermore, dorsal root ganglion neurons that had been treated with a CKII inhibitor or transfected with the L1 mutants showed impaired ability to extend axons on an L1 substrate but not on other control substrates. These results demonstrate the novel role of CKII in L1-mediated axon elongation and stress the importance of functional linkage between L1 phosphorylation and L1 trafficking in migrating growth cones.

Aryl hydrocarbon receptor (AhR) is a transcription factor that works as a dioxin receptor and is also involved in various physiological phenomena, including development and cell proliferation. Here, we show that the Ga 13 signal destabilizes AhR by promoting the ubiquitination of AhR. Ga 13 interacts directly with AhR-interacting protein (AIP) and inhibits the interaction between AhR and AIP, a crucial interacting protein of AhR. Strikingly, a reporter gene assay and a quantitative reverse transcription-PCR analysis indicate that the Ga 13 signal shows a potent inhibitory effect on the ligand-induced transcriptional activation of AhR. Ga 13 results in the nuclear translocation of AhR in a ligand-independent manner. However, in the presence of active Ga 13 , AhR fails to form the active transcriptional complex. Taken together, we propose a new negative regulation of dioxin signalling by the G protein.