Despite of their developmental importance and the striking number of visual defects that arise from aberrant neural crest development, little is known of the molecular signals that control their migration during eye development or during the regeneration of the cornea. In the periocular region, neural crest cells express Neuropilin-1 (Npn-1) prior to their migration into the eye, while the lens expresses SemaphorinSA (SemaSA). Interestingly, Npn-1 and SemaSA are also differentially expressed as the cornea regenerates during wound healing. However the role of Npn-1/SemaSA signaling during eye development and wound healing has not been examined. Therefore it is critical to identify and characterize the role of guidance molecules that regulate the migration of periocular neural crest cells and their keratocyte progeny in the cornea. This project will study the role of Npn-1/SemaSA interactions during neural crest migration in the eye. There will be special emphasis on the invasion of periocular neural crest into the eye rudiment, their interaction with the lens and differentiation into corneal endothelium and stroma, and relationship to neural crest-derived keratocytes during regeneration of the cornea. This proposal combines classical embryological techniques with molecular biology and video microscopy to visualize neural crest migration in real-time and examine the role of Npn-1 and SemaSA during these events. Experiments will be carried out in chick and mouse, as each model offers unique experimental advantages. The following specific aims will be performed: 1) Examine the role of Npn-1/SemaSA interactions during neural crest contribution to the cornea in vivo and in vitro. 2) Examine the migration of neural crest cells into the cornea by in vivo time-lapse video microscopy. 3) Characterize the expression of Npn-1 and SemaSA during mouse eye development. 4) Examine the role of Npn-1/SemaSA interactions during mouse eye development. 5) Examine the role of Npn-1/SemaSA interactions during cornea wound healing. Significance: The proposed research aims to provide an insight into the role of guidance molecules during neural crest migration into the eye during development and of their keratocyte progeny during cornea regeneration. Understanding the role of guidance molecules during -these processes may provide clues on how the migration pattern of neural crest cells is disrupted in congenital eye disorders. ? ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Career Transition Award (K99)
Project #
5K99EY018050-02
Application #
7492065
Study Section
Special Emphasis Panel (ZEY1-VSN (03))
Program Officer
Shen, Grace L
Project Start
2007-09-01
Project End
2009-08-31
Budget Start
2008-09-01
Budget End
2009-08-31
Support Year
2
Fiscal Year
2008
Total Cost
$88,830
Indirect Cost
Name
California Institute of Technology
Department
Type
Schools of Arts and Sciences
DUNS #
009584210
City
Pasadena
State
CA
Country
United States
Zip Code
91125
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Spurlin 3rd, James; Lwigale, Peter (2013) A technique to increase accessibility to late-stage chick embryos for in ovo manipulations. Dev Dyn 242:148-54
McKenna, Chelsey C; Munjaal, Ravi P; Lwigale, Peter Y (2012) Distinct roles for neuropilin1 and neuropilin2 during mouse corneal innervation. PLoS One 7:e37175
McKenna, Chelsey C; Lwigale, Peter Y (2011) Innervation of the mouse cornea during development. Invest Ophthalmol Vis Sci 52:30-5
Lwigale, Peter Y; Bronner-Fraser, Marianne (2009) Semaphorin3A/neuropilin-1 signaling acts as a molecular switch regulating neural crest migration during cornea development. Dev Biol 336:257-65
Lwigale, Peter Y; Bronner-Fraser, Marianne (2007) Lens-derived Semaphorin3A regulates sensory innervation of the cornea. Dev Biol 306:750-9