Retinoid storage in the egg of reptiles and birds.

Storage of retinal has been confirmed in eggs from a range of anamniotic vertebrates (teleosts and amphibians) and an ascidian, but the retinoid-storage state in eggs of oviparous amniotic vertebrates (reptiles and birds) has yet to be clarified in detail. We studied four reptilian and five avian species and found that retinal was commonly stored in their egg yolk. Furthermore, retinal was the major retinoid in reptilian eggs, with only low levels of retinol, whereas significant amounts of retinol as well as retinal were stored in avian eggs. In both reptilian and avian eggs, retinal was commonly bound to proteins, which were assumed to be homologous to the proteins that bind retinal in the eggs of anamniotic vertebrates. Despite the common storage state of retinal, retinol would be bound to different proteins. In the reptilian eggs, retinol was found in the yolk-granule fraction, which also contained retinal. However, retinol in avian eggs was found largely in the yolk-plasma fraction, separate from retinal. These results suggest that retinol storage in avian eggs acquired after the divergence of birds from the reptiles, while retinal storage was acquired before the appearance of the vertebrates, and has subsequently been conserved during evolution of oviparous vertebrates.

Repulsive and attractive semaphorins cooperate to direct the navigation of cardiac neural crest cells.

The cardiac neural crest, a subpopulation of the neural crest, contributes to the cardiac outflow tract formation during development. However, how it follows the defined long-range migratory pathway remains unclear. We show here that the migrating cardiac neural crest cells (NCCs) express Plexin-A2, Plexin-D1 and Neuropilin. The membrane-bound ligands for Plexin-A2, Semaphorin (Sema)6A and Sema6B, are expressed in the dorsal neural tube and the lateral pharyngeal arch mesenchyme (the NCC "routes"). Sema3C, a ligand for Plexin-D1/neuropilin-1, is expressed in the cardiac outflow tract (the NCC "target"). Sema6A and Sema6B repel neural crest cells, while Sema3C attracts neural crest cells. Sema6A and Sema6B repulsion and Sema3C attraction are diminished either when Plexin-A2 and Neuropilin-1, or when Plexin-D1, respectively, are knocked down in NCCs. When RNAi knockdown diminishes each receptor in NCCs, the NCCs fail to migrate into the cardiac outflow tract in the developing chick embryo. Furthermore, Plexin-A2-deficient mice exhibit defects of cardiac outflow tract formation. We therefore conclude that the coordination of repulsive cues provided by Sema6A/Sema6B through Plexin-A2 paired with the attractive cue by Sema3C through Plexin-D1 is required for the precise navigation of migrating cardiac NCCs.

FARP2 triggers signals for Sema3A-mediated axonal repulsion.

Sema3A, a prototypical semaphorin, acts as a chemorepellent or a chemoattractant for axons by activating a receptor complex comprising neuropilin-1 as the ligand-binding subunit and plexin-A1 as the signal-transducing subunit. How the signals downstream of plexin-A1 are triggered upon Sema3A stimulation, however, is unknown. Here we show that, in the presence of neuropilin-1, the FERM domain-containing guanine nucleotide exchange factor (GEF) FARP2 associates directly with plexin-A1. Sema3A binding to neuropilin-1 induces the dissociation of FARP2 from plexin-A1, resulting in activation of FARP2's Rac GEF activity, Rnd1 recruitment to plexin-A1, and downregulation of R-Ras. Simultaneously, the FERM domain of FARP2 sequesters phosphatidylinositol phosphate kinase type I isoform PIPKIgamma661 from talin, thereby inhibiting its kinase activity. These activities are required for Sema3A-mediated repulsion of outgrowing axons and suppression of neuronal adhesion. We therefore conclude that FARP2 is a key molecule involved in the response of neuronal growth cones to class-3 semaphorins.