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The Journal of Neuroscience, January 1, 1998, 18(1):184-194
Laminin Directs Growth Cone Navigation via Two Temporally and
Functionally Distinct Calcium Signals
Thomas B.
Kuhn1,
Cheri V.
Williams2,
Ping
Dou2, and
S. B.
Kater3
Departments of 1 Biochemistry and Molecular Biology and
2 Anatomy and Neurobiology, Colorado State University, Fort
Collins, Colorado 80523, and 3 Department of Anatomy and
Neurobiology, University of Utah School of Medicine, Salt Lake City,
Utah 84132
During development, growth cones navigate to their targets via
numerous interactions with molecular guidance cues, yet the mechanisms
of how growth cones translate guidance information into navigational
decisions are poorly understood. We have examined the role of
intracellular Ca2+ in laminin (LN)-mediated growth
cone navigation in vitro, using chick dorsal root
ganglion neurons. Subsequent to contacting LN-coated beads with
filopodia, growth cones displayed a series of stereotypic changes in
behavior, including turning toward LN-coated beads and a phase of
increased rates of outgrowth after a pause at LN-coated beads. A
pharmacological approach indicated that LN-mediated growth cone turning
required an influx of extracellular Ca2+, likely in
filopodia with LN contact, and activation of calmodulin (CaM).
Surprisingly, fluorescent Ca2+ imaging revealed no
LN-induced rise in intracellular Ca2+ in filopodia
attached to their parent growth cone. However, isolation of filopodia
by laser-assisted transection unmasked a rapid, LN-specific rise in
intracellular Ca2+ (+73 ± 11 nM).
Additionally, a second, sustained rise in intracellular Ca2+ (+62 ± 8 nM) occurred in
growth cones, with a distinct delay 28 ± 3 min after growth cone
filopodia contacted LN-coated beads. This delayed, sustained
Ca2+ signal paralleled the phase of increased rates
of outgrowth, and both events were sensitive to the inhibition of
Ca2+/CaM-dependent protein kinase II (CaM-kinase II)
with 2 µM KN-62. We propose that LN-mediated growth cone
guidance can be attributed, in part, to two temporally and functionally
distinct Ca2+ signals linked by a signaling cascade
composed of CaM and CaM-kinase II.
Key words:
Ca2+; laminin; growth cone; filopodia; pathfinding; calmodulin; Ca2+/calmodulin-dependent
protein kinase II
Copyright © 1998 Society for Neuroscience 0270-6474/98/181184-11$05.00/0
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