Research Paper Volume 12, Issue 7 pp 6436—6455

Sevoflurane inhibits neuronal migration and axon growth in the developing mouse cerebral cortex

Figure 4. Netrin-1/Dcc deficiency displayed a similar defect in neuronal migration and axon outgrowth as Nova2 deficiency induced by dual sevoflurane exposure. (A) Dcc shRNA (sh3) reliably reduces the Dcc expression. (B) Quantification of the protein expressions of Dcc relative to Actin (F = 10.57, P = 0.0226*, N = 3, one-way ANOVA). (C) Netrin-1 shRNA (sh3) reliably reduces the Netrin-1 expression. (D) Quantification of the protein expressions of Netrin-1 relative to Actin (F = 7.084, P = 0.0445*, N = 3, one-way ANOVA. (E) Dcc and Netrin-1 knockdown both significantly inhibited neuronal migration cortex of offspring mice. (F) Quantification of GFP+ cells at different positions in different groups. Compared to the Con-sh group, the Dcc-sh group had significantly larger fractions of neurons in the IZ (40.23 ± 1.20208%, P = 0.0005***, N = 3, Student’s t-test) and the VZ/SVZ (3.93 ± 2.50315%, P = 0.13, N = 3, Student’s t-test), and a significantly smaller fraction of neurons in the UpCP (28.335 ± 1.9728%, P = 0.0434*, N = 3, Student’s t-test). (G) Quantification of GFP+ cells at different positions in different groups. Compared to the Con-sh group, the Netrin-1-sh group had significantly larger fractions of neurons in the IZ (42.665 ± 6.73872%, P = 0.0239*, N = 3, Student’s t-test) and the VZ/SVZ (5.335 ± 1.0818%, P = 0.8521, N = 3, Student’s t-test), and a significantly smaller fraction of neurons in the UpCP (26.57 ± 2.0223%, P = 0.0037**, N = 3, Student’s t-test). (H) Both Dcc and Netrin-1 knockdown decreased axon length of neurons in primary cultured mouse cortical neurons. (I) The statistical results for the axon length in Dcc-sh RNA group (P = 0.0054**, Student’s t-test) and Netrin-1-shRNA group (P = 0.0044**, Student’s t-test). Scale bars = 100 μm; approximately 70 cells from three independent experiments were counted during the statistical analysis. *P<0.05. **P<0.01, ***P<0.001.****P< 0.0001.