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Lamellipodia mediate the heterogeneity of central olfactory ensheathing cell interactions

Abstract

The growth and guidance of primary olfactory axons are partly attributed to the presence of olfactory ensheathing cells (OECs). However, little is understood about the differences between the subpopulations of OECs and what regulates their interactions. We used OEC-axon assays and determined that axons respond differently to peripheral and central OECs. We then further purified OECs from anatomically distinct regions of the olfactory bulb. Cell behaviour assays revealed that OECs from the olfactory bulb were a functionally heterogeneous population with distinct differences which is consistent with their proposed roles in vivo. We found that the heterogeneity was regulated by motile lamellipodial waves along the shaft of the OECs and that inhibition of lamellipodial wave activity via Mek1 abolished the ability of the cells to distinguish between each other. These results demonstrate that OECs from the olfactory bulb are a heterogeneous population that use lamellipodial waves to regulate cell–cell recognition.

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Acknowledgments

This work was supported by a grant from the National Health and Medical Research Council to J.S. and B.K. (grant number 511006), by funding to the National Centre for Adult Stem Cell Research from the Australian Government Department of Health and Aging (A.M.S.) and by an Australian Postgraduate Award to L.W.

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Correspondence to James A. St John.

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Movie 1: Central OECs adhere to one another during cell-cell contact. Contact between the leading edge of one OEC and the shaft of another OEC resulted in the formation of a lamellipodial wave. A subsequent retraction of the shaft of one OEC is observed allowing the two leading edges to merge and cell-cell adhesion takes place. Time is recorded in min, sec and msec. Selected frames shown in Fig. 2A. (MPEG 566 kb)

Movie 2: Central OECs repel from each other during cell-cell contact. The leading edge of an OEC moved toward a second OEC which extended a lamellipodial wave with filopodia that interacted with the leading edge. Despite a brief interaction, the leading edge retracted within minutes. Time is recorded in hr, min and sec. Selected cropped frames are shown in Fig. 2B. (MPEG 1262 kb)

Movie 3: Central OECs also display non-responsive behaviour during cell-cell contact. As OECs crossed over, the lamellipodial waves that emerged on the shaft of the cell collapsed following contact with the adjacent cell. A lamellipodial wave extended a single filopodium which contacted the shaft of the second OEC resulting in collapse of the wave. Both OECs continued to exist without obvious adhesion or repulsion occurring. Time is recorded min, sec and msec. Selected cropped frames are shown in Fig. 2C. (MPEG 792 kb)

Movie 4: Motile lamellipodial waves travel in both an anterograde and/or retrograde direction along a central OEC. Several lamellipodial waves appear along the shaft of an isolated OEC process. The initial wave travelled in a retrograde direction while the second wave developed and travelled anterograde prior to merging with a third lamellipodial wave. Merged wave subsequently moved retrograde toward the cell body. Time is recorded in hr, min and sec. Selected cropped frames are shown in Fig. 3A. (MPEG 1328 kb)

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Windus, L.C.E., Lineburg, K.E., Scott, S.E. et al. Lamellipodia mediate the heterogeneity of central olfactory ensheathing cell interactions. Cell. Mol. Life Sci. 67, 1735–1750 (2010). https://doi.org/10.1007/s00018-010-0280-3

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Keywords

  • Olfactory bulb
  • Axon
  • Regeneration
  • Glia
  • Mek1