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Laminin-332 coordinates mechanotransduction and growth cone bifurcation in sensory neurons

Abstract

Laminin-332 is a major component of the dermo-epidermal skin basement membrane and maintains skin integrity. The transduction of mechanical force into electrical signals by sensory endings in the skin requires mechanosensitive channels. We found that mouse epidermal keratinocytes produce a matrix that is inhibitory for sensory mechanotransduction and that the active molecular component is laminin-332. Substrate-bound laminin-332 specifically suppressed one type of mechanosensitive current (rapidly adapting) independently of integrin-receptor activation. This mechanotransduction suppression could be exerted locally and was mediated by preventing the formation of protein tethers necessary for current activation. We also found that laminin-332 could locally control sensory axon branching behavior. Loss of laminin-332 in humans led to increased sensory terminal branching and may lead to a de-repression of mechanosensitive currents. These previously unknown functions for this matrix molecule may explain some of the extreme pain experienced by individuals with epidermolysis bullosa who are deficient in laminin-332.

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Figure 1: Keratinocyte-derived matrix suppresses mechanotransduction currents.
Figure 2: Laminin-332 reproduces the suppression of rapidly adapting mechanosensitive currents.
Figure 3: Laminin-332–containing matrix does not support tether formation and exerts its effect independent of integrin receptors.
Figure 4: Laminin-332 suppression of the rapidly adapting mechanosensitive current is local, not global.
Figure 5: Differential growth behavior on laminin and laminin-332.
Figure 6: Laminin-332 markedly alters the network structure of the matrix.
Figure 7: Human laminin-332 deficiency sensitizes mechanotransduction.
Figure 8: Altered sensory afferent branching in the skin of laminin-332–deficient individuals.

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Acknowledgements

We thank members of the Lewin laboratory for advice and comments on the manuscript. H. Thränhardt and A. Wegner provided excellent technical support. We thank R. Bönsch for performing the western blot for laminin-332. We also thank M. Bastmeyer and A. von Philipsborn for their help in establishing the micro-contact printing method in the laboratory. We are grateful to B. Erdmann for help and advice with the TEM experiments. This work was supported by grants from the Deutsche Forschungsgemeinschaft (G.R.L. and M.K.), a Network Epidermolysis Bullosa grant from the Federal Ministry of Education and Research (BMBF, L.B.-T.) and a von Humboldt fellowship (J.H.).

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J.H., L.-Y.C. and K.P. performed the electrophysiology experiments and L.-Y.C. carried out the TEM analysis. B.E.O. and M.K. provided laminin-111/Nidogen complexes and recombinant β3 and γ2. K.P. carried out micro-contact printing with L.-Y.C. and performed time-lapse experiments. K.P. carried out AFM experiments. Neurite outgrowth assays and human skin immunocytochemistry were performed by K.P. with help from N.D. and Y.A.B.S. L.B.-T. provided and characterized the human keratinoctyes. L.-Y.C., J.H., K.P. and G.R.L. planned the experiments and analyzed data. G.R.L. and J.H. wrote the paper.

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Correspondence to Jing Hu or Gary R Lewin.

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Supplementary Video 1

Neurite outgrowth over laminin/laminin cross-hatched pattern. (AVI 38049 kb)

Supplementary Video 2

Neurite outgrowth over laminin/laminin-332 cross-hatched pattern. (AVI 25049 kb)

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Chiang, LY., Poole, K., Oliveira, B. et al. Laminin-332 coordinates mechanotransduction and growth cone bifurcation in sensory neurons. Nat Neurosci 14, 993–1000 (2011). https://doi.org/10.1038/nn.2873

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