{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T01:05:40Z","timestamp":1789693540701,"version":"4.0.1"},"posted":{"date-parts":[[2025,12,1]]},"group-title":"Cell Biology","reference-count":58,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T00:00:00Z","timestamp":1764547200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,29]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  The \u201cbrain-eating amoeba\u201d\n                  <jats:italic>Naegleria fowleri<\/jats:italic>\n                  dwells in ponds where it normally feeds on bacteria, but if it enters the brain it can cause a deadly infection. To establish infection,\n                  <jats:italic>N. fowleri<\/jats:italic>\n                  must migrate through different environments\u2014along olfactory axons, through openings in the cribriform plate, and within brain tissue\u2014yet how it does so remains unknown. As a model for\n                  <jats:italic>N. fowleri<\/jats:italic>\n                  migration within these environments, we examine how its non-pathogenic relative,\n                  <jats:italic>Naegleria gruberi<\/jats:italic>\n                  , navigates environments of distinct geometries. We show that\n                  <jats:italic>Naegleria<\/jats:italic>\n                  uses both actin-rich protrusions and membrane blebs to crawl across or between flat surfaces. We also explore how\n                  <jats:italic>Naegleria<\/jats:italic>\n                  interact with narrow channels and find that, unlike\n                  <jats:italic>Dictyostelium<\/jats:italic>\n                  amoebae that we show frequently disengage from channel interfaces,\n                  <jats:italic>Naegleria<\/jats:italic>\n                  amoebae probe channels until they enter. Once inside,\n                  <jats:italic>Naegleria<\/jats:italic>\n                  crawls quickly (&gt;50 \u03bcm\/min) and unidirectionally over long distances (&gt;1 mm) using only bleb-based motility. We also introduced\n                  <jats:italic>Naegleria<\/jats:italic>\n                  to granular hydrogel matrices that mimic pond sediments and found that cells readily enter and migrate through these three-dimensional matrices using both blebs and lamellar protrusions. Although cells in matrices showed lower persistence at short timescales, longer time scales correlate with increased persistence, suggesting\n                  <jats:italic>Naegleria<\/jats:italic>\n                  cells may retain memory of past orientation. We propose that pond life may select for three behaviors that prime\n                  <jats:italic>Naegleria<\/jats:italic>\n                  for pathogenesis: memory-guided motility that would facilitate exploration of sinus cavities, confinement-seeking (\u201cclaustrophilia\u201d) that would promote entry into narrow passages along olfactory axons, and persistent bleb-based migration that would allow rapid transit along axons to the brain.\n                <\/jats:p>","DOI":"10.1101\/2025.11.30.691445","type":"posted-content","created":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T12:30:10Z","timestamp":1764592210000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Naegleria<\/i>\n                  amoebae seek confinement and crawl persistently through narrow spaces"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9985-9522","authenticated-orcid":false,"given":"Katrina B.","family":"Velle","sequence":"first","affiliation":[{"name":"Department of Biology, University of Massachusetts Dartmouth, North Dartmouth, MA 02747, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9220-2256","authenticated-orcid":false,"given":"Meera","family":"Ramaswamy","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA"},{"name":"Princeton Center for Complex Materials, Princeton University, Princeton, NJ, 08544"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Babak Vajdi","family":"Hokmabad","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering, Princeton University, Princeton NJ 08544, 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