Asymmetric redistribution of GABA receptors during GABA gradient sensing by nerve growth cones analyzed by single quantum dot imaging

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 104; no. 27; pp. 11251 - 11256
• Main Authors: Bouzigues, Cédric, Morel, Mathieu, Triller, Antoine, Dahan, Maxime
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 03.07.2007; National Acad Sciences
• Subjects: Amino acids; Amoeba; Animals; Biochemistry; Biological Sciences; Biological Transport, Active - physiology; Calcium; Cell Polarity - physiology; Cells, Cultured; Chemotaxis - physiology; Cholinergic receptors; Cytoskeleton; gamma-Aminobutyric Acid - metabolism; Growth Cones - chemistry; Growth Cones - metabolism; Growth Cones - physiology; Intracellular Fluid - chemistry; Intracellular Fluid - metabolism; Intracellular Fluid - physiology; Kinetics; Molecules; Nervous system; Neurons; Neurons - chemistry; Neurons - cytology; Neurons - metabolism; Physical Sciences; Quantum Dots; Rats; Receptors; Receptors, GABA-A - metabolism; Second Messenger Systems - physiology; Signal amplification; T cell receptors; Trajectories
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0702536104

During development of the nervous system, the tip of a growing axon, the growth cone (GC), must respond accurately to stimuli that direct its growth. This axonal navigation depends on extracellular concentration gradients of numerous guidance cues, including GABA. GCs can detect even weak directional signals, yet the mechanisms underlying this sensitivity remain unclear. Past studies in other eukaryotic chemotactic systems have pointed to the role of the spatial reorganization of the transduction pathway in their sensitive response. Here we have developed a single-molecule assay to observe individual GABAA receptors (GABAARs) in the plasma membrane of nerve GCs subjected to directional stimuli. We report that in the presence of an external GABA gradient GABAARs redistribute asymmetrically across the GC toward the gradient source. Single-particle tracking of GABAARs shows that the redistribution results from transient interactions between the receptors and the microtubules. Moreover, the relocalization is accompanied by an enhancement in the asymmetry of intracellular calcium concentration. Altogether, our results reveal a microtubule-dependent polarized reorganization of chemoreceptors at the cell surface and suggest that this polarization serves as an amplification step in GABA gradient sensing by nerve GCs.