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Chapter 16

Ephrins and Eph Receptor Tyrosine Kinases in Synapse Formation


Catherine E. Krull and Daniel J. Liebl

Abstract Here, we discuss what is known about the function of ephrins and Eph receptors in synapse formation in the peripheral nervous system and central nervous system. Ephrins have been shown to be present and functional at synapses at the neuromuscular junction and in the brain; evidence at the neuromuscular junction implicates ephrins in the topographic mapping of synapses on certain muscles. Also, certain Eph receptors also function in synapse formation but less information is known about them including their distribution and function. In addition, future directions are defined. Together, these data implicate ephrins, as well as Ephs, strongly in synaptic development. Keywords Synaptogenesis Eph family PNS CNS

16.1 Introduction
Eph receptor tyrosine kinases (RTKs) and their ligands, the ephrins, were first recognized for their roles in axon pathfinding, cell migration, and angiogenesis (Krull et al. 1997, Wang and Anderson 1997, Smith et al. 1997, Adams et al. 1999, Helmbacher et al. 2000, Eberhart et al. 2000, Eberhart et al. 2002, Kullander et al. 2001, Eberhart et al. 2004, Sahin et al. 2005). However, these molecules are now thought to play central roles in the formation of synapses as well (Donoghue et al. 1996, Feng et al. 2000, Murai et al. 2003, Grunwald et al. 2004, Rodenas-Ruano et al. 2006, Bourgin et al. 2007). Here, in addition to providing pertinent background, recent data will be reviewed that define a key function for these molecules in the formation of synapses in the peripheral nervous system (PNS) and the central nervous system (CNS). Information on other aspects of Ephephrin function can be found in several reviews (Pasquale
C.E. Krull (*) Biologic and Materials Sciences, University of Michigan, 5211 Dental, 1011 N. University Ave., Ann Arbor, MI 48109-1078, USA e-mail: krullc@umich.edu

M. Hortsch, H. Umemori (eds.), The Sticky Synapse, DOI 10.1007/978-0-387-92708-4_16, Springer ScienceBusiness Media, LLC 2009

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2008, Arvanitis and Davy 2008, Himanen et al. 2007, Kullander and Klein 2002, Murai and Pasquale 2004, Wilkinson 2001).

16.2 The Eph Family: Description


Several members of the Eph family of receptor tyrosine kinases (RTKs) and their membrane-associated ligands, the ephrins, have been implicated in patterning in the developing nervous system and angiogenesis (Wilkinson 2001, Table 16.1). Based on their ligand-binding specificities, the Eph RTKs are subdivided into two subclasses, EphAs, which interact primarily with GPIlinked ephrins (i.e., ephrin-As), and EphBs, which bind to and activate transmembrane ephrins (i.e., ephrin-Bs) (Gale et al. 1996, but see Himanen et al. 2004). The extracellular region of the Eph receptors includes an N-terminal globular domain that binds ephrin, a cysteine-rich domain, and two fibronectin type III repeats (Yamaguchi and Pasquale 2004). The tyrosine kinase domain is
Table 16.1 Known vertebrate Eph family members Members of the Eph family Eph receptor tyrosine kinases Ephrin ligands EphA1 Ephrin-A1 EphA2 Ephrin-A2 EphA3 Ephrin-A3 EphA4 Ephrin-A4 EphA5 Ephrin-A5 EphA6 Ephrin-A6 EphA7 EphA8 EphA9 EphA10 EphB1 Ephrin-B1 EphB2 Ephrin-B2 EphB3 Ephrin-B3 EphB4 EphB5 EphB6 Originally, the Eph family was divided into two subgroups: EphA receptor tyrosine kinases (RTKs) bound primarily ephrin-As; EphB RTKs bound ephrin-Bs (Eph Nomenclature Committee). However, there is a significant amount of cross talk between the two subgroups in biological events, so that EphA RTKs interact with ephrin-Bs, and EphB RTKs interact with ephrin-As. Investigators are urged to examine whether an Eph or ephrin from a different subgroup is involved in the biological event being examined. Molecule names emphasized by an outline font format are implicated in synapse formation.

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located below a juxtamembrane region, followed by a sterile-a-motif (SAM) and a type II PSD-95/Disc large/ZO-1 (PDZ)-binding motif at the carboxyl terminus (for review, Kullander and Klein 2002, Fig. 16.1). Importantly, Eph receptors can undergo homo- or heterodimerization (Freywald et al. 2002), which is regulated directly by the extracellular cysteine-rich domain (Lackmann et al. 1998) and the SAM motif (Stapleton et al. 1999, Thanos et al. 1999) or indirectly through PDZ protein interactions (Fanning and Anderson 1999). Localization of ephrins to the cell surface is critical for their ability to activate their receptors (Davis et al. 1994). Because of their membrane association, Eph RTKephrin interactions require cellcell contact that initiates signals from both the ligand- and the receptor-expressing cells. The requirement for bidirectional signaling has been demonstrated best for ephrin-Bs and EphB2 (Henkemeyer et al. 1996, Cowan and Henkemeyer. 2001, Lu et al. 2001). Studying ephrin-A signaling has proved more difficult, as they do not have direct contact with the cytoplasm because of their GPI anchor. However, previous studies suggest that signaling is mediated by the localization of

Fig. 16.1 General protein domain organization of Ephrins and Eph receptor tyrosine kinases. A schematic diagram that shows ephrins (top cell) interacting with a canonical Eph receptor tyrosine kinase (bottom cell), including important domains. Note that in most systems, Eph receptors and ephrins are expressed in the same cell; the function of this colocalization is not well understood

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GPI-anchored ephrins in cholesterol-rich microdomains in the cell membrane (Davy et al. 1999, Sargiacomo et al. 1993, Shenoy-Scaria et al. 1994). How signaling works exactly to alter an axons response from adhesion to repulsion is not well understood, but is thought to involve trans-endocytosis and enzymatic cleavage (Hattori et al. 2000, Zimmer et al. 2003, Marston et al. 2003, Irie et al. 2005, Lin et al. 2008). Several mouse mutants have been generated that lack particular Ephs or ephrins (Kullander et al. 2001, Egea et al. 2005, Henkemeyer et al. 1996, Feng et al. 2000, Grunwald et al. 2004). Mutant analyses have shown that certain Eph receptors and ephrins are required for the formation of axonal commissures, patterning of the retinocollicular system, synapse formation, and synaptic plasticity (Henkemeyer et al. 1996, Cheng et al. 1995, Feng et al. 2000, Kullander et al. 2001, McLaughlin et al. 2003, Grunwald et al. 2004, Mendes et al. 2006, Rodenas-Ruano et al. 2006). There are additional factors, including Ret and GDNF, that cooperate with Ephs and ephrins for axon pathfinding. However, exactly how they function together remains elusive (Kramer et al. 2006). Furthermore, it is not known whether Ephs/ephrins work with other axon guidance molecules, such as semaphorins/neuropilins and plexins (Chauvet et al. 2007, Huber et al. 2005, Giger et al. 2000); it is becoming more and more pressing to know how the growth cone of a neuron responds to multiple guidance cues as it navigates to its targets and forms synapses.

16.3 Ephrins Control Neuromuscular Topography and Synapse Formation in the PNS
Before any molecule is implicated in synapse formation, it must be localized to either presumptive synapses or their targets (i.e., muscles or other neurons) (Murai and Pasquale 2004). Localization to synapses or to targets is determined by many methods including antibody staining (Garcia et al. 2007, Hu et al. 2007, Kania et al. 2000). Needless to say, in situ hybridization usually provides limited information, as the transcript of interest localizes to the nucleus and this approach cannot provide information regarding the localization of the protein to the growth cone or developing synapses. In the PNS, using subtractive hybridization, Donoghue and colleagues found that ephrin-A5 (AL-1) is expressed by rostral skeletal muscles and inhibits the outgrowth from caudal (lumbar) neurons in a mouse model system (Donoghue et al. 1996). Wang, Chadaram and colleagues explored this finding further, revealing that spinal cord neurites maintain their preference for muscle membranes derived from their corresponding axial level using in vitro stripe assays (Wang et al. 2001; Chadaram et al. 2007). For the first time, these authors showed that topographic outgrowth of motor neurons occurs within a single motor pool onto a single target

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muscle in vitro. Overexpression of ephrin-A5 degrades the topographic specificity of the gluteus maximus muscle (Lampa et al. 2004). Feng and his colleagues established that ephrin-A5 was required in mouse for selective synaptogenesis in the PNS in vivo (Feng et al. 2000). Their results showed that motor axons and their target muscles possess complementary molecular labels that vary with rostrocaudal position and that, therefore, bias synaptic connections in favor of matched partners (i.e., motor axons, muscle). Feng and colleagues first demonstrated this pattern by comparing ephrin-A RNAs expression between rostral versus caudal muscles, using RT-PCR and Northern blot analysis. They also assayed the cellular source of ephrin-As using fusion protein staining. While this approach does not identify which ephrin protein is expressed, it did demonstrate whether ephrin-A proteins were expressed collectively by myotubes. Importantly, no ephrin-Bs were expressed by adult PNS synapses (Yamaguchi and Pasquale 2004), indicating that ephrin-As are present selectively at the right time and place to influence synapse formation. Feng and colleagues then evaluated the differential sensitivity of motor neurons from rostral versus caudal levels to ephrin-A5 expressed by fibroblasts. Their results showed that ephrin-A5 inhibited growth from both rostral and caudal neurons, but that caudal neurons were on average more sensitive to ephrin-A5. Also, shallow gradients of ephrin-A5 were formed and the outgrowth of rostral versus caudal neurons was measured. Here too, the growth of rostral versus caudal neurons was inhibited, but caudal neurons were better able to detect differences in ephrin concentration. The authors then generated mice where ephrin-A5 was overexpressed in certain muscle fibers. The rostrocaudal axis of the gluteus muscle pool was dramatically degraded in mice with excess ephrin-A5 compared to wild-type mice. Abolishing the gradient of ephrin-A5 on muscle led to disruptions in the neuromuscular map, demonstrating that ephrin-A5 can affect specific synaptic connections. To determine whether endogenous ephrins were required for this process, the authors examined diaphragm muscles from mice that lacked ephrin-A2 and ephrin-A5. Here, topographic mapping of synapses onto muscle was also defective. Collectively, these data indicate that muscle-derived ephrins act in selective synaptic connections between nerve and muscle partners (Feng et al. 2000). How ephrins act specifically to regulate nervemuscle synapses is obviously the next question. As for Eph receptors, EphA4 has been knocked out by two research groups (Helmbacher et al. 2000, Dottori et al. 1998). One set of mice that lack EphA4 has defects in the dorsoventral choice made by motor axons (Helmbacher et al. 2000). However, it is unclear whether synapse formation is also affected in these mice. Moreover, the general organization of muscle appears normal in these mice. Mice that lack other EphAs have also been reported, but it is not known whether there are defects in neuromuscular synaptogenesis (Irie et al. 2008, Shintani et al. 2006).

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16.4 Ephs and Ephrins in Synapse Formation in the CNS


Although there is sufficient evidence that ephrins act during synapse formation in the PNS, there is some controversy regarding their activities in the CNS. The controversy stems from whether Ephs and ephrins are involved in synapse formation or synaptic function and which receptor or ligand acts in specific cell types. Grunwald and colleagues recently examined the localization of ephrin-B2 and its function in synaptic plasticity in the hippocampus (Grunwald et al. 2001, 2004). Using in situ hybridization and antibody staining, Grunwald and colleagues found that ephrin-B2 and ephrin-B3 are localized postsynaptically to CA3CA1 synapses in the hippocampus. To directly visualize ephrin-B2 at CA1CA3 synapses, these authors then performed electron microscopy using a specific antibody against ephrin-B2. Collectively, these results showed that ephrin-B2 protein localizes postsynaptically. Rodenas-Ruano and colleagues also demonstrated that ephrin-B3 is localized to both axons and dendrites and is located to the postsynaptic membrane in CA3CA1 synapses (Rodenas-Ruano et al. 2006). One recent report identifies ephrin-B2 protein in the cell bodies of neurons in the adult mouse brain, suggesting it may have roles in the formation of axonalsomatic inhibitory synapses (Migani et al. 2007). Subsequently, Grunwald and colleagues conditionally removed ephrin-B2 from the postnatal forebrain and examined synapse formation using electron microscopy (Grunwald et al. 2004). Interestingly, they found no difference in synapse number or morphology compared to control mice. In addition, the authors examined hippocampal neurons from mice that lacked EphA4, a highaffinity receptor for ephrin-B2, and control mice and found no difference in synaptogenesis. Together, these findings indicate that ephrin-B2 is not required for synapse formation in the hippocampus. Grunwald and colleagues then analyzed basal synaptic transmission in both mutant and control embryos (Grunwald et al. 2004). Again, no difference in the extracellular recordings of the CA3CA1 pathway in the hippocampus of mutant and control mice was apparent. However, a strong reduction in LTP was observed after the application of theta-burst stimulation, an experimental paradigm that is used to induce long-lasting LTP. Together, these data show that ephrin-B2 is required for hippocampal LTP. In addition, using similar approaches, these authors and others examined the contribution of ephrin-B3 to LTP and found that it too was required for early stages of LTP (Grunwald et al. 2004, Rodenas-Ruano et al. 2006). Moreover, LTD was also affected in these mice that lacked ephrin-B2 and ephrin-B3, compared to controls. Finally, due to its known role as a high-affinity receptor for ephrin-B2 and ephrin-B3, EphA4 mutant mice were examined for their role in LTP (Gale et al. 1996). Based on localization and functional analyses, EphA4 appears to be a critical interaction partner of ephrin-B2 and ephrin-B3 in the early phase of LTP (Grunwald et al. 2001, Grunwald et al. 2004). Interestingly, mice that lack the

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signaling portion of ephrin-B3, but retain their ability to activate Eph receptors, do not exhibit a similar reduction in LTP, suggesting that these mechanisms may involve Eph receptor signaling and/or multiple ephrins (Rodenas-Ruano et al. 2006). Using three different in vivo approaches, Lim and colleagues examined the effect of the presynaptic ephrin-B reverse intracellular signaling on synapse formation in the retinotectal system (Lim et al. 2008). First, soluble fusion proteins (EphB2-Fc and ephrin-B1-Fc) were infused into the brain, followed by assaying synaptic morphology. Second, plasmid constructs that encoded ephrin-B1 or a mutated form of ephrin-B1 were electroporated into the retina, followed by an analysis of presynaptic development. Third, plasmids that encoded a mutated form of EphB2 were electroporated into the tectum, to interfere with endogenous EphBephrin-B signaling, and the effects on retinotectal synapses were examined. Their results show that activating ephrin-B1 signaling in retinal ganglion cells promotes the formation of presynaptic specializations without affecting overall axon growth. In contrast, reducing reverse signaling slows this process. In addition, these authors demonstrated that EphBephrin-B interactions affect synaptic function, including synaptic plasticity. There is other tantalizing data that suggest that EphB receptors participate in synaptogenesis between neurons. Henkemeyer and colleagues have reported that EphB2 acts postsynaptically by modulating glutamate receptors (Henkemeyer et al. 2003). There is additional data that support the postsynaptic function of EphB2. In cultured hippocampal neurons, EphB2 interacts directly with NMDA receptors and thus modulates the influx of calcium into postsynaptic neurons. In addition, postsynaptic EphB2 receptors interact with the PDZ-domain protein GRIP at the mossy fiber synapses and mediate long-term potentiation (LTP). EphB receptors are known to co-distribute with a nicotinic acetylcholine receptor subtype (a7-nAChR) and regulate nicotinic downstream signaling in isolated parasympathetic ganglia neurons in vitro (Liu et al. 2008). Signaling is enhanced by the activation of EphB receptors interacting with a7-nAChRs on the neurons. However, interactions between EphB receptors/ephrin-B1 and a7-nAChRs are not direct and it is not known whether EphB/ ephrin-B1 may also have a role in cell death in this system. Henkemeyer, Hoogenraad and their colleagues have shown that EphB (EphB1, EphB2, EphB3) receptors are necessary for the formation of dendritic spines, small protrusions on the dendrites surface that receive most of excitatory synaptic input (Henkemeyer et al. 2003, Hoogenraad et al. 2005). Their data strongly indicate that EphB receptors, to varying degrees, are involved in the morphogenesis of dendritic spines and synapse formation in the mouse hippocampus. Previously, EphB receptors were shown to be likely molecules that transduced extracellular signals into the dendrite to induce spine morphogenesis (Ethell et al. 2001). Indeed, EphBs are found in the postsynaptic structures of dendrites (Torres et al. 1998, Buchert et al. 1999) and ephrin-Bs have been localized to axons (Henkemeyer et al. 1996). At least four members of the

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Eph receptors (EphB1, EphB2, EphB3, EphB4) are expressed in the hippocampus (Liebl et al. 2003). Furthermore, presynaptic specializations are induced in HEK-293 cells that overexpress EphB2 in co-cultured cortical neurons (Kayser et al. 2006). In addition, there is a link between ephrin-B/EphB signaling and Rho-GEF kalirin-7 in spine morphogenesis in cultured hippocampal neurons (Penzes et al. 2003, see Table 16.1). EphB receptors also promote the formation of spines by activating Rho family GEFs, including kalirin-7 (see above) and Tiam1 (Penzes et al. 2003, Tolias et al. 2007). Rho GTPases, including Rac1, RhoA, and Cdc42, are key regulators of the actin cytoskeleton and are known to play critical roles in spine morphogenesis. Rho family GTPases act as molecular switches, cycling between an active GTP-bound state and an inactive GDP-bound state. The activity of Rho family GTPases is controlled tightly by guanine nucleotide exchange factors (GEFs). Tolias and colleagues determined that the Rac1 GEF Tiam1 mediates EphB receptor-dependent dendritic spine development (Tolias et al. 2007). Activation of EphB receptors by ephrin-B ligands induces the tyrosine phosphorylation of Tiam1 and recruitment of Tiam1 to EphB complexes that contain NMDA-type glutamate receptors. Knockdown of Tiam1 or expression of a dominant-negative Tiam1 blocks signaling, and most importantly, dendritic spine formation induced by ephrin-B1 stimulation. Indeed, Dalva and colleagues demonstrated previously that ephrin-B activation of EphBs leads to the recruitment of NMDA receptors to an EphB-containing complex and influences the number of pre- and postsynaptic specializations (Dalva et al. 2000). More recently, Kayser and colleagues showed that loss of EphBs results in reduced filopodial motility and abnormal synapse formation in cultured neurons and brain slices (Kayser et al. 2008). Rescue experiments during different developmental times show that EphBs are required for synaptogenesis, but only when filopodia are plenty and motile (Kayser et al. 2008). In contrast, EphA receptors appear to promote spine retraction (Murai et al. 2003). How do EphA receptors perform this task mechanistically? Fu and colleagues recently showed that Cdk5 regulates EphA4-mediated spine retraction through an ephexin1-dependent mechanism (Fu et al. 2007). Blocking Cdk5 activity inhibits ephrin-A1-triggered spine retraction and a reduction in the frequency of mESPCs at hippocampal synapses. Activating EphA4 results in the recruitment of Cdk5 to EphA4, leading to the activation of Cdk5. Then, together, these factors activate ephexin1, a Rho-GEF, leading to spine retraction. In another example, EphA4 seems to regulate dendritic spine remodeling by affecting b1-integrin signaling pathways (Bourgin et al. 2007). The model proposed by Bourgin and colleagues is that EphA4 interferes with integrin signaling pathways that stabilize dendritic spines, thus modulating synaptic interactions with the extracellular milieu. The situation may be more complex than first appreciated. Murai and colleagues have shown that astrocytes express ephrin-A3, a membrane-bound ligand for EphA4, and that it regulates the morphology of dendritic spines in the hippocampus (Murai et al. 2003). Their data suggest that local activation of

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EphA receptors on dendritic spines occurs by astrocytic ephrin-A3. Time-lapse imaging studies showed that dendritic protrusions are highly active (Dailey and Smith, 1996) and the idea has emerged that a subset of motile protrusions are stabilized and transformed into stable spines with cellcell contacts. Murai and colleagues suggested that local astrocytic contacts may be important for maintaining stable spines and regulate the maturation of these dendritic protrusions. Nishida and Okabe further examined the formation of spines, using pyramidal neurons in hippocampal slices, and tested the role of ephrins and Ephs in this process (Nishida and Okabe 2007). The authors observed frequent contact between astrocytes and dendritic protrusions. Furthermore, the authors found that perturbation of Rac1 signaling in astrocytes to suppress astrocytic motility results in the development of abnormal and thin dendritic protrusions and that blocking Eph and ephrin signaling reduces the lifetime of newly generated dendritic protrusions with astrocytic contacts. Lauterbach and Klein showed that EphB2 receptor-containing vesicles are released by hippocampal neurons to co-cultured glial cells, suggesting that glial cells can transendocytose full-length EphB2 from neighboring neurons (Lauterbach and Klein 2006). Together, these results suggest that glial cells express ephrin-A3 as well as EphB2 receptors and thus should be considered as important players at the synapse.

16.5 Summary and Future Directions


Ephrins and Eph receptors play a crucial role in synapse formation in the PNS and CNS. Interestingly, the analyses of mouse mutants indicate that many members of this family are required for synapse formation. However, a substantial body of data is still missing: 1. How exactly do ephrins and their Eph receptors work at a mechanistic level to regulate synapse formation? 2. How do ephrins and Eph receptors, expressed by the same cell, function together? Do all receptors have the capacity to work in cis and in trans? 3. What is the role of trans-endocytosis in the process of synapse formation? 4. Are ephrins involved in dendrite versus axon targeting (i.e., neuronal polarity)? Answers to these questions await future investigations in this very open and fertile area.

References
Adams RH, Wilkinson GA, Weiss C et al. (1999) Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. Genes Dev 13:295306

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Arvanitis D and Davy A (2008) Eph/ephrin signaling: networks. Genes Dev 22:416429 Buchert M, Schneider S, Meskenaite V et al. (1999) The junction-associated protein AF-6 interacts and clusters with specific Eph receptor tyrosine kinases at specialized sites of cellcell contact in the brain. J Cell Biol 144:361371 Bourgin C, Murai KK, Richter M et al. (2007) The EphA4 receptor regulates dendritic spine remodeling by affecting beta1-integrin signaling pathways. J Cell Biol 178:12951307 Chadaram SR, Laskowski MB and Madison RD (2007) Topographic specificity within membranes of a single muscle detected in vitro. J Neurosci 27:1393813948 Chauvet S, Cohen S, Yoshida Y et al. (2007) Gating of Sema3E/PlexinD1 signaling by neuropilin-1 switches axonal repulsion to attraction during brain development. Neuron 56:807822 Cheng HJ, Nakamoto M, Bergemann AD et al. (1995) Complementary gradients in expression and binding of ELF-1 and Mek4 in development of the topographic retinotectal projection map. Cell 82:371381 Cowan CA and Henkemeyer M (2001) The SH2/SH3 adaptor Grb4 transduces B-ephrin reverse signals. Nature 413:174179 Dailey ME and Smith SJ (1996) The dynamics of dendritic structure in developing hippocampal slices. J Neurosci 16:29832994 Dalva MB, Takasu MA, Lin MZ et al. (2000) EphB receptors interact with NMDA receptors and regulate excitatory synapse formation. Cell 103:945956 Davis S, Gale NW, Aldrich TH et al. (1994) Ligands for the Eph-related receptor tyrosine kinases that require membrane attachment or clustering for activity. Science 266:816819 Davy A, Gale NW, Murray EW et al. (1999) Compartmentalized signaling by GPI-anchored ephrin-A5 requires the Fyn tyrosine kinase to regulate cellular adhesion. Genes Dev 13:31253135 Donoghue MJ, Merlie J and Sanes JR (1996) The Eph kinase ligand AL-1 is expressed by rostral muscles and inhibits outgrowth from caudal neurons. Mol Cell Neurosci 8:185198 Dottori M, Hartley L, Galea M et al. (1998) EphA4 (Sek1) receptor tyrosine kinase is required for the development of the corticospinal tract. PNAS 95:1324813253 Eberhart J, Swartz M, Koblar SA et al. (2000) Expression of EphA4, ephrin-A2 and ephrinA5 during axon outgrowth to the hindlimb indicates potential roles in pathfinding. Dev Neurosci 22:237250 Eberhart J, Barr J, OConnell S et al. (2004) Ephrin-A5 exerts positive or inhibitory effects on distinct subsets of EphA4-positive motor neurons. J Neurosci 24:10701078 Eberhart J, Swartz ME, Koblar SA et al. (2002) EphA4 constitutes a population-specific guidance cue for motor neurons. Dev Biol 247:89101 Egea J, Nissen UV, Dufour A et al. (2005) Regulation of EphA4 kinase activity is required for a subset of axon guidance decisions suggesting a key role for receptor clustering in Eph function. Neuron 47:515528 Ethell IM, Irie F, Kalo MS et al. (2001) EphB/syndecan-2 signaling in dendritic spine morphogenesis. Neuron 31:10011013 Fanning AS and Anderson JM (1999) PDZ domains: fundamental building blocks in the organization of protein complexes at the plasma membrane. J Clin Invest 103:767772 Feng G, Laskowski MB, Feldheim DA et al. (2000) Roles for ephrins in positionally selective synaptogenesis between motor neurons and muscle fibers. Neuron 25:295306 Freywald A, Sharfe N and Roifman CM (2002) The kinase-null EphB6 receptor undergoes transphorylation in a complex with EphB1. J Biol Chem 277:38233828 Fu W-Y, Chen Y, Sahin M et al. (2007) Cdk5 regulates EphA4-mediated dendritic spine retraction through an ephexin1-dependent mechanism. Nat Neurosci 10:6776 Gale NW, Flenniken A, Compton DC et al. (1996) Elk-3, a novel transmembrane ligand for the Eph family of receptor tyrosine kinases, expressed in embryonic floor plate, roof plate, and hindbrain segments. Oncogene 13:13431352

16

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Garcia SM, Casanueva MO, Silva MC et al. (2007) Neuronal signaling modulates protein homeostasis in Caenorhabditis elegans post-synaptic muscle cells. Genes Dev 21:30063016 Giger RJ, Cloutier JF, Sahay A et al. (2000) Neuropilin-2 is required in vivo for selective axon guidance responses to secreted semaphorins. Neuron 25:2941 Grunwald IC, Korte M, Wolfer D et al. (2001) Kinase-independent requirement of EphB2 receptors in hippocampal synaptic plasticity. Neuron 32:10271040 Grunwald IC, Korte M, Adelmann G et al. (2004) Hippocampal plasticity requires postsynaptic ephrin-Bs. Nat Neurosci 7:3340 Hattori M, Osterfield M and Flanagan JG (2000) Regulated cleavage of a contact-mediated axon repellent. Science 289:13601365 Helmbacher F, Schneider-Maunoury S, Topilko P et al. (2000) Targeting of the EphA4 tyrosine kinase receptor affects dorsal/ventral pathfinding of limb motor axons. Development 127:33133324 Henkemeyer M, Orioli D, Henderson JT et al. (1996) Nuk controls pathfinding of commissural axons in the mammalian central nervous system. Cell 86:3546 Henkemeyer M, Itkis OS, Ngo M et al. (2003) Multiple EphB receptor tyrosine kinases shape dendritic spines in the hippocampus. J Cell Biol 163:13131326 Himanen JP, Saha N and Nikolov DB (2007) Cell-cell signaling via Eph receptors and ephrins. Curr Opin Cell Biol 19:534542 Himanen JP, Chumley MJ, Lackmann M et al. (2004) Repelling class discrimination: ephrinA5 binds to and activates EphB2 receptor signaling. Nat Neurosci 7:501509 Hoogenraad CC, Milstein AD, Ethell IM et al. (2005) GRIP1 controls dendrite morphogenesis by regulating EphB receptor trafficking. Nat Neurosci 8:906915 Hu JY, Chen Y and Schacher S (2007) Multifunctional role of protein kinase C in regulating the formation and maturation of specific synapses. J Neurosci 27:1171211724 Huber AB, Kania A, Tran TS et al. (2005) Distinct roles for secreted semaphoring signaling in spinal motor axon guidance. Neuron 48:949964 Irie F, Okuno M, Pasquale EB et al. (2005) EphrinB-EphB signaling regulates clathrinmediated endocytosis through tyrosine phosphorylation of synaptojanin 1. Nat Cell Biol 7:454456 Irie F, Okumo M, Matsumoto K et al. (2008) Heparan sulfate regulates ephrin-A3/EphA receptor signaling. PNAS 34:1230712312 Kayser MS, McClelland AC, Hughes EG et al. (2006) Intracellular and trans-synaptic regulation of glutamatergic synaptogenesis by EphB receptors. J Neurosci 26:1215212164. Kayser MS, Nolt MJ and Dalva MB (2008) EphB receptors couple dendritic filopodia motility to synapse formation. Neuron 59:5669 Kania A, Johnson RL and Jessell TM (2000) Coordinate roles for LIM homeobox genes in directing dorsoventral trajectory of motor axons in the vertebrate limb. Cell 102:161173 Kramer ER, Knott L, Su F et al. (2006) Cooperation between GDNF/Ret and ephrinA/ EphA4 signals for motor axon pathway selection in the limb. Neuron 50:3547 Krull CE, Lansford R, Gale NW et al. (1997) Interaction of Eph-related receptors and ligands confer rostrocaudal pattern to trunk neural crest migration. Curr Biol 7:571580 Kullander K and Klein R (2002) Mechanisms and functions of Eph and ephrin signaling. Nat Rev Mol Cell Biol 3:475486 Kullander K, Mather NK, Diella F et al. (2001) Kinase-dependent and kinase-independent functions of EphA4 receptors in major axon tract formation in vivo. Neuron 29:7384 Lackmann M, Oates AC, Dottori M et al. (1998) Distinct subdomains of the EphA3 receptor mediate ligand binding and receptor dimerization. J Biol Chem 273:2022820237 Lampa SJ, Potluri S, Norton AS et al. (2004) Ephrin-A5 overexpression degrades topographic specificity in the mouse gluteus maximus muscle. Dev Brain Res 153:271274 Lauterbach J and Klein R (2006) Release of full-length EphB2 receptors from hippocampal neurons to cocultured glial cells. J Neurosci 26:1157511581

344

C.E. Krull and D.J. Liebl

Liebl DJ, Morris CJ, Henkemeyer M et al. (2003) mRNA expression of ephrins and Eph receptor tyrosine kinases in the neonatal and adult mouse central nervous system. J Neurosci Res 71:722 Lim BK, Matsuda N and Poo MM (2008) Ephrin-B reverse signaling promotes structural and functional synaptic maturation in vivo. Nat Neurosci 11:160169 Lin KT, Sloniowski S, Ethell DW et al. (2008) Ephrin-B2 induced cleavage of EphB2 receptor is mediated by matrix metalloproteinases to trigger cell repulsion. J Biol Chem. (Available online) Liu Z, Conroy WG, Stawicki TM et al. (2008) EphB receptors co-distribute with a nicotinic receptor subtype and regulate nicotinic downstream signaling in neurons. Mol Cell Neurosci 38:236244 Lu Q, Sun EE, Klein RS et al. (2001) Ephrin-B reverse signaling is mediated by a novel PDZRGS protein and selectively inhibits G protein-coupled chemoattraction. Cell 105:6979 Marston DJ, Dickinson S and Nobes CD (2003) Rac-dependent trans-endocytosis of ephrinBs regulates Eph-ephrin contact repulsion. Nat Cell Biol 5:879888 McLaughlin T, Hindges R, Yates PA et al. (2003) Bifunctional action of ephrin-B1 as a repellent and attractant to control bidirectional branch extension in dorsal-ventral retinotopic mapping. Development 130:24072418 Mendes SW, Henkemeyer M and Liebl DJ (2006) Multiple Eph receptors and B-class ephrins regulate midline crossing of corpus callosum fibers in the developing mouse forebrain. J Neurosci 26:882892 Migani P, Bartlett C, Dunlop S et al. (2007) Ephrin-B2 immunoreactivity distribution in the adult mouse brain. Brain Res 1182:6072 Murai KK, Nguyen LN, Irie F et al. (2003) Control of hippocampal dendritic spine morphology through ephrin-A3/EphA4 signaling. Nat Neurosci 6:153160 Murai KK and Pasquale EB (2004) Eph receptors, ephrins and synaptic function. The Neuroscientist 10:304314 Nishida H and Okabe S (2007) Direct astrocytic contacts regulate local maturation of dendritic spines. J Neurosci 27:331340 Pasquale EB (2008) Eph-ephrin bidirectional signaling in physiology and disease. Cell 133:3852 Penzes P, Beeser A, Chernoff J et al. (2003) Rapid induction of dendritic spine morphogenesis by trans-synaptic ephrinB-EphB receptor activation of the Rho-GEF kalirin. Neuron 37:263274 Rodenas-Ruano A, Perez-Pinzon MA, Green EJ et al. (2006) Distinct roles for ephrin-B3 in the formation and function of hippocampal synapses. Dev Biol 292:3445 Sahin M, Greer PL, Lin MZ et al. (2005) Eph-dependent tyrosine phosphorylation of ephexin1 modulates growth cone collapse. Neuron 46:191204 Sargiacomo M, Sudol M, Tang Z et al. (1993) Signal transducing molecules and glycosylphosphatidylinositol-linked proteins form a caveolin-rich insoluble complex in MDCK cells. J Cell Biol 122:789807 Shenoy-Scaria AM, Dietzen DJ, Kwong J et al. (1994) Cysteine3 of Src family protein tyrosine kinase determines palmitoylation and localization in caveolae. J Cell Biol 126:353363 Shintani T, Ihara M, Sakuta H et al. (2006) Eph receptors are negatively controlled by protein tyrosine phosphatase receptor type O. Nat Neurosci 9:761769 Smith A, Robinson V, Patel K et al. (1997) The EphA4 and EphB1 receptor tyrosine kinases and ephrin-B2 ligand regulate targeted migration of brachial neural crest cells. Curr Biol 7:561570 Stapleton D, Balan I, Pawson T et al. (1999) The crystal structure of an Eph receptor SAM domain reveals a mechanism for modular dimerization. Nat Struct Biol 6:4449 Tolias KF, Bikoff JB, Kane CG et al. (2007) The Rac1 guanine nucleotide exchange factor Tiam1 mediates EphB receptor-dependent dendritic spine development. PNAS 104:72657270

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Ephrins and Eph Receptor Tyrosine Kinases

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Torres R, Firestein BL, Dong H et al. (1998) PDZ proteins bind, cluster and synaptically colocalize with Ephs and their ephrin ligands. Neuron 21:12271229 Thanos CD, Faham S, Goodwill KE et al. (1999) Monomeric structure of the human EphB2 sterile alpha motif domain. J Biol Chem 274:3730137306 Wang HU and Anderson DJ (1997) Eph family transmembrane ligands can mediate repulsive guidance of trunk neural crest migration and motor axon outgrowth. Neuron 18:383396 Wang H, Chadaram SR, Norton AS et al. (2001) Development of inhibition by ephrin-A5 on outgrowth of embryonic spinal motor neurites. J Neurobiol 47:233243 Wilkinson DG (2001) Multiple roles of EPH receptors and ephrins in neural development. Nat Rev Neurosci 2:155164 Yamaguchi Y and Pasquale EB (2004) Eph receptors in the adult brain. Curr Opin Neurobiol 14:288296 Zimmer M, Palmer A, Kohler J et al. (2003) EphB-ephrinB bi-directional endocytosis terminates adhesion allowing contact mediated repulsion. Nat Cell Biol 5:869878