Axon Guidance

Multiple decision points along a growing axons trajectory Different types of axon guidance cues:
Contact mediated - requires direct contact by growth cone Long range - growth cone responds to chemical gradient

Growth cones respond to these different types of cues at different points along their trajectories to navigate toward their ultimate targets Sperrys chemoaffinity hypothesis

A number discrete steps and decision points along the way A. To leave the retina at the otic nerve head (ONH) B. To cross (or not) at the optic chiasm (and where) C. A/P and D/V positions

The retinotectal projection: a point-to-point topographic map inverted over A/P and D/V axes
Retina Tectum

Tessier-Lavigne, Cell 1995

Roger Sperry: rotation of retina - RGCs still make connections according to their original (intrinsic) positions
Normal Eye rotated 180o

Sperry: Chemoaffinity Hypothesis (1963) 1. Each position in the optic tectum has a unique molecular address or molecular tag 2. Each retinal ganglion cell (RGC) has a unique set of receptors for these tags (i.e., an identity), resulting in a positiondependent response dependent on differential affinities or differential intracellular responses 3. Unlikely to be unique molecules for each position (would be insufficient information in the genome to wire up entire brain this way) - rather, information is probably encoded in orthogonal gradients (A/P, D/V) => Positional identities of axons and targets are matched up to establish the point-to-point topographic map

Axons are guided by different types of guidance mechanisms

Extracellular matrix adhesion Cell surface adhesion Fasciculation Contact inhibition

Contact mediated
Chemoattraction

Chemorepulsion

Long Range

Axon growth cone is a sensory-motor structure that recognizes and responds to guidance cues

Santiago Ramn y Cajal 1852-1934

Structure of the growth cone

lamellipodium filopodium

Actin is concentrated in filopodia and lamellipodia

Microtubules are concentrated in the central core of the growth cone

Filopodia sensory ability of the GC.

Growth cone extension

Filopodia extended

Microtubules from central core advance

Cytoplasm collapses to create new segment of axon

Extracellular Matrix Molecules (Contact Mediated)

Laminins: major components of basal laminae and account for much of the axon outgrowth promoting ability of the extracellular matrix. Heterotrimers of related !,",and # subunits (5 !, 4 " and 3 # genes at least 11 trimers has been identified). Integrins: Expressed on the growth cone, interacts with laminin (16 ! and 8 " genes). Heterodimers of ! and ", recognize different laminins.

Cell Adhesion Molecules (Contact Mediated)

Cadherin superfamily

Immunoglobulin superfamily

Binds Ca2+

Binds catenins, links to cytoskeletons

Contact-Mediated Repulsion
Friedrich Bonhoeffers stripe assay demonstrates sensitivity of posterior (temporal) retinal axons to a repellent activity in posterior tectal membranes

Repulsive signaling mediated by the Ephrins (ligand) and Eph receptors

Wilkinson, Nat. Rev. Neurosci. 2001

Manipulations of Ephrin Expression Alter Topographic Map

Normal: A -> P P -> A

Overexpress ephrin in spots in tectum: A -> P P avoids areas w/ephrin

Reduced ephrin in tectum: A -> P P -> A, P

Chemorepulsion (Long Range)

Growth cone collapse upon exposure to a chemorepellent

Growth cone collapse from point source (gradient) leads to axon turning away from chemorepellent source

The Semaphorins and their receptors

Chemoattraction (Long Range)

Netrin-1: secreted by the floorplate gradient guides axon turning

Netrin-1 receptor mediating attractive response: DCC - vertebrates, is evolutionarily conserved = unc-40 - C. elegans (worms) = frazzled - Drosophila (flies)

Netrin-1 can act as both chemoattractant and chemorepellent

Response to netrin-1 is dependent on two factors 1) Netrin receptors: DCC/unc-40/frazzled: attractive response DCC/unc-40 + unc5: repulsive response

2) cAMP levels and protein kinase A (PKA) activation

low cAMP High cAMP

Robo receptors transduce a midline repulsive signal encoded in slit proteins and silence netrin-mediated attraction