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Variability in Membrane Continuity Between Schwann Cells and Neurons

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Abstract

Both Schwann cells (SCs) and neurons dynamically expand and contract their plasma membrane during their extension of projections and movement. However, these cell types have very different motility profiles and physiological function. We developed methods to measure the correlated movement of regions of plasma membrane, based on quantitative analysis of the movement of fluorescently labeled wheat-germ agglutinin (WGA) bound to the extracellular membrane. WGA trajectories were compared between SCs and neurons isolated from neonatal Sprague–Dawley rats, using both cross-correlation and regression analysis. Schwann cellular membranes exhibited significantly higher correlation (42.37 ± 5.87%, mean ± SEM) compared to neurons (24.51 ± 3.52%). Additionally, Schwann cellular membranes were more mobile (−0.165 ± 0.099 μm/min average velocity) compared to neurons (0.052 ± 0.032 μm/s). Comparison of both cell types upon establishment of contact with another neuron failed to identify any difference with the non-contacting state. Our results are suggestive of a role for forces generated by mobility on the biomechanical continuity of plasma membrane. Such forces are likely to interact with factors, including the cytoskeletal framework and adhesion proteins. This work has implications for interactions of neurons and SCs during development and neuronal regeneration.

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Acknowledgments

We gratefully acknowledge funding from the National Science Foundation (CBET 1212301) and the Maryland Stem Cell Research Fund.

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Authors and Affiliations

  1. Fischell Department of Bioengineering, University of Maryland, College Park, MD, 20742, USA

    James M. Love, Gunja K. Pathak, Joshua Chetta & Sameer B. Shah

  2. Departments of Orthopaedic Surgery and Bioengineering, University of California, San Diego, La Jolla, CA, 92093, USA

    Sameer B. Shah

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Correspondence to Sameer B. Shah.

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Associate Editor William H Guilford oversaw the review of this article.

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Supplemental Fig. 1 Control Experiments. Growth cone ruffling was observed in non-contacting neurons (A) as an indication of overall neuronal health, scale bar 10 μm. Axonal blebbing (B) was observed in a limited number of neurons. These neurons were excluded from the study, scale bar 10 μm. High velocity membrane movement was observed in axons that were given 4 h to recover following WGA application (C), scale bar 20 μm. To exclude internalized membrane, the 4-h incubation was removed from the protocol. Cumulative histogram of individual positive particle velocities on neurons (D) *p < 0.0001 (v. no incubation, Kolmogorov–Smirnov test). Confocal imaging of fluoresce revealed the absence of internalized WGA following initial incubation (E), scale bar 20 μm.

Supplemental Fig. 2 Particle Velocity Histograms by cell type and contact state. Additional histograms of positive (A) and negative (C) particle velocities (analogous to Figs. 3f and 3g). Focused characterization of the velocities of particles in regions within 30 μm of the edge (B and D) yielded little difference. Significant differences (p < 0.05, Kolmogorov–Smirnov) are as follow: (A) NC vs. SCC, NC vs. SCNC, NNC vs. SCC, NNC vs. SCNC; (B) NC vs. SCNC, NNC vs. SCC, NNC vs. SCNC, SCC vs. SCNC; (C) NC vs. SCC, NC vs. SCNC, NNC vs. SCC, NNC vs. SCNC, SCC vs. SCNC; (D) NC vs. SCC, NC vs. SCNC, NNC vs. SCC, NNC vs. SCNC.

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Love, J.M., Pathak, G.K., Chetta, J. et al. Variability in Membrane Continuity Between Schwann Cells and Neurons. Cel. Mol. Bioeng. 5, 450–462 (2012). https://doi.org/10.1007/s12195-012-0250-y

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