Figure 3

Blocking Cb2r prevents the CBN-induced deficits in morphology and survival. (A) Images of embryos at 2 dpf in the presence and absence of Cb1r and Cb2r antagonists with CBN. N = 3 experiments. (B) Bar graph shows the mean body length (mm) of embryos for one set of antagonists (AM251 and AM630 for Cb1r and Cb2r respectively) in vehicle control (n = 57), 3 mg L⁻¹ CBN (n = 29), AM251 10 nm + 3 mg L⁻¹ CBN (n = 21), AM630 10 µm + 3 mg L⁻¹ CBN (n = 21) and AM251 10 nm + AM630 10 µm + 3 mg L⁻¹ CBN (n = 36) respectively at 2 dpf (N = 3 experiments). (C) Bar graph shows the mean body length (mm) of embryos for the second set of antagonists (Cp94 and JTE907 for Cb1r and Cb2r respectively). The bars represent vehicle control (n = 57), 3 mg L⁻¹ CBN (n = 29), CP94 10 nm + 3 mg L⁻¹ CBN (n = 31), JTE907 10 µm + 3 mg L⁻¹ CBN (n = 234) and Cp94 10 nm + JTE907 10 µm + 3 mg L⁻¹ CBN (n = 31) at 2 dpf. N = 3 experiments. (D) Line graph showing the percentage of embryos that survived within the first 5 days of development following co-exposure of the first set of antagonists and CBN. Treatments are vehicle control, 3 mg L⁻¹ CBN, AM251 10 nm + 3 mg L⁻¹ CBN, AM630 10 µm + 3 mg L⁻¹ CBN and AM251 10 nm + AM630 10 µm + 3 mg L⁻¹ CBN (N = 4 experiments and n = 25 embryos for each experiment). (E) Line graph showing the percentage of embryos that survived within the first 5 days of development following co-exposure of the second set of antagonists and CBN. Treatments are vehicle control, 3 mg L⁻¹ CBN, CP94 10 nm + 3 mg L⁻¹ CBN, JTE907 10 µm + 3 mg L⁻¹ CBN and Cp94 10 nm + JTE907 10 µm + 3 mg L⁻¹ CBN (N = 4 experiments and n = 25 embryos for each treatment). (F) Bar graphs show the mean heart rate in beats per minute (bpm) at 2 dpf for vehicle control (n = 35), 3 mg L⁻¹ CBN (n = 59), AM251 10 nm + 3 mg L⁻¹ CBN (n = 32), AM630 10 µm + 3 mg L⁻¹ CBN (n = 33) and AM251 10 nm + AM630 10 µm + 3 mg L⁻¹ CBN (n = 20) respectively. N = 4 experiments. (G) Bar graphs show heart rate as beats per minute (bpm) at 2 dpf for vehicle control (n = 35), 3 mg L⁻¹ CBN (n = 59), CP94 10 nm + 3 mg L⁻¹ CBN (n = 21), JTE907 10 µm + 3 mg L⁻¹ CBN (n = 25) and Cp94 10 nm + JTE907 10 µm + 3 mg L⁻¹ CBN (n = 20) respectively. N = 4 experiments. Groups which share the same letter(s) of the alphabet are not statistically different from one another; p < 0.05.