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Tracing Complex Neural Crest Cell Behaviors in the Embryo using Photoactivation of KikGR
Author(s) -
Kulesa Paul,
Teddy Jessica,
Stark Danny,
Smith Sarah,
McLennan Rebecca
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.101.2
Subject(s) - neural crest , biology , embryo , microbiology and biotechnology , population , live cell imaging , cell , embryonic stem cell , confocal , anatomy , green fluorescent protein , neuroscience , genetics , optics , physics , demography , sociology , gene
Key to our understanding of how molecular mechanisms modulate cell migratory patterns in the embryo is information about cell behaviors at precise times and spatial locations. However, it has been challenging to selectively mark subgroups of migratory cells in vivo without disruption to the local microenvironment. We apply a combination of photoactivation cell labeling using KikGR, and confocal time‐lapse imaging to investigate cell behaviors within the migratory front and trailing subgroups of chick neural crest (NC), a highly invasive pluripotent embryonic cell population. KikGR is a fluorescent protein that is electroporated into NC cells and contains a green chromophore that is photoconvertible to red following 405 nm laser excitation, thus allowing less invasive optical precision to fluorescently mark single and small groups of cells. We find that the spatial order of cranial NC cell emergence translates into a distal‐to‐proximal invasion of the target branchial arches. Lead and trailing NCCs display similar average cell speeds and directionalities. Surprisingly, we find that lead NCCs proliferate along the migratory route and grow to outnumber trailing NCCs by nearly 3 to 1. Our results suggest a model in which lead and trailing NCC behaviors are distinct and coordinated to maintain a spatially‐ordered invasion of the branchial arches. Research support is from the Stowers Institute.