FIGURE 1.

Effect of Lhx8 overexpression on cholinergic neurons. After 3 DIV, rat E18.5 primary septum neurons were infected with AAV-GFP or AAV-Lhx8, and those cells were fixed at 8 DIV (A, C, and D) or exposed to Krebs-like buffer containing 6 mm K⁺ (B). A, representative images of septum neurons infected with AAV-GFP and immunostained with anti-MAP2 (panels a–c) or anti-choline transporter (panels d–f and d′–f′) or anti-TrkA (panels g–i, and g′–i′) antibodies. Virtually all MAP2-positive cells expressed GFP. B, ACh release evaluated after 30 min of stimulation with 6 mm K⁺. Data are expressed as a percentage of the ACh concentration released from AAV-GFP-infected neurons. C and D, number of choline transporter (CHT)- and TrkA-positive cells counted. No significant difference in the number of cholinergic neurons was observed. Scale bar, 30 μm. Data are presented as the mean ± S.E. n = 4; *, p < 0.05; **, p < 0.01; analysis of variance with post hoc Dunnett's multiple comparison test.