Pax6 +/+ or
Pax6 +/ − corneal epithelial cells were cultured in either control medium or media containing 50 μg/mL of ConA, WGA, or DBA for 30 minutes. ConA and WGA were chosen, because they have been shown to be localized to the leading edge of healing rat corneal epithelia.
32 A single linear scratch of up to 200 μm was made in the epithelial sheet, and the rate of wound healing was measured at 37°C. Wild-type cells started to heal almost immediately (
first 2 hours: rate of wound healing = 30.4 ± 5.1 μm/hour;
n = 26; and
2 to 6 hours after wounding: rate of wound healing = 29.8 ± 2.7 μm/hour;
n = 26). In contrast,
Pax6 +/ − cells hardly migrated at all within the first 2 hours of wounding, but from 2 to 6 hours after wounding, migrated at a rate that was not significantly different (
t-test:
P = 0.086) from wild-type (
first 2 hours: rate of wound healing = 5.9 ± 2.1 μm/hour;
2 to 6 hours after wounding: rate of wound healing = 36.7 ± 3.1 μm/hour;
n = 26). Addition of ConA or WGA to the medium of wild-type cells reduced the immediate wound-healing cell migration to levels not significantly different from untreated
Pax6 +/ − cells (ConA = 4.2 ± 1.5 μm/hour,
n = 12; WGA = 5.6 ± 3.2 μm/hour,
n = 8). The migration of ConA-treated cells remained significantly slower than untreated wild-type cells in the next 4 hours (9.8 ± 2.0 μm/hour,
n = 12;
t-test:
P < 0.0001). WGA-treated cells started to migrate more quickly once they had overcome their first 2-hour block, although this was of marginal significance (19.1 ± 4.5 μm/hour;
n = 8;
t-test:
P = 0.030). The migration of WGA-treated wild-type cells 2 to 6 hours after wounding was not significantly different from that of either untreated wild-type cells (
t-test:
P = 0.062) or ConA-treated cells (
t-test:
P = 0.089). These data are summarized in
Figure 6 .