14-644 Sigma-AldrichEphA3 Protein, active, 10 µg

Stimulation of CD28 alone has been shown to regulate cytokine gene transcription and expression of the type 1 insulin-like growth factor receptor (IGF-1R) in lymphocytes. In this study, the ephrin receptor tyrosine kinase ephA3, was identified as a new CD28-responsive gene in Jurkat cells by using a human cytokine/receptor array. EphA3 was not detected in normal peripheral T cells, in any subset of thymus-derived developing T cells, or in Hodgkin's lymphoma. However, contrary to previous findings, EphA3 was detected in a panel of T-cell lymphomas. Stimulation of Jurkat cells with ephrin-A5 resulted in loss of cell adhesion to fibronectin and recruitment of the adapter protein CrkII to EphA3. Interestingly, EphA3 expression in CD28-stimulated Jurkat cells was enhanced by IGF-1 or by overexpression of the IGF-1R, and was suppressed by anti-IGF-1R blocking antibodies. The data suggest that CD28- and IGF-1-regulated expression of EphA3 is associated with adherence and that it may be involved in the motility of malignant T cells.

The control of cell movement during development is essential for forming and stabilizing the spatial organization of tissues and cell types. During initial steps of tissue patterning, distinct regional domains or cell types arise at appropriate locations, and the movement of cells is constrained in order to maintain spatial relationships during growth. In other situations, the guidance of migrating cells or neuronal growth cones to specific destinations underlies the establishment or remodeling of a pattern. Eph receptor tyrosine kinases and their ephrin ligands are key players in controlling these cell movements in many tissues and at multiple stages of patterning.

Axons of retinal ganglion cells exhibit a specific pattern of connections with the brain. Within each visual nucleus in the brain, retinal connections are topographic such that axons from neighboring ganglion cells have neighboring synapses. Research is beginning to shed light on the mechanisms responsible for development of topographic connections in the visual system. Much of this research is focused on the axonal connections of the retina with the tectum. In vivo and in vitro experiments indicate that the pattern of retinotectal connections develops in part due to positional labels carried by the growing retinal axons and by the tectal cells. Evidence suggests that gradients of Eph receptor tyrosine kinases serve as positional labels on the growing retinal axons, and gradients of ligands for these receptors serve as positional labels in the tectum. Blocking expression of EphA3, a receptor tyrosine kinase, in the developing retina resulted in disruption of the topography of the retinotectal connections, further supporting the role of these, molecules. Although positional labels appear to be important, other mechanisms must also be involved. The initial pattern of retinotectal connections lacks the precision seen in the adult. The adult pattern of connections arises during development by activity dependent refinement of a roughly ordered prepattern. The refinement process results in elimination of projections to the wrong side of the brain, to non-visual nuclei and to inappropriate regions within a nucleus. Blocking NMDA receptors during the period of refinement preserved anomalous retinotectal projections, which suggests that elimination of these projections is mediated by NMDA receptors. Furthermore, tectal cells normally express high levels of nitric oxide synthase (NOS) during the period of refinement, and blocking nitric oxide (NO) synthesis also preserved inappropriate projections. Thus, both NMDA receptors and NO appear to be involved in refinement. Blocking NMDA receptor activation reduced NOS activity in tectal cells, which suggests the possibility that NO is the downstream mediator of NMDA function related to refinement. A quantitative comparison of blocking NMDA receptors, NO synthesis or both showed that all three treatments have comparable effects on refinement. This indicates that the role of NMDA receptor activation relative to refinement may be completely mediated through nitric oxide. Quantitative analysis also suggests that other mechanisms not involving NMDA receptors or NO must be involved in refinement. Other mechanisms appear to include cell death.