Fig. 5: Startle by abrupt air-puff stimulation induces astrocytic Ca²⁺ elevations and LC/NA axon multipeak Ca²⁺ signals without a cAMP surge.

a Illustration of air-puff stimulation under two-photon microscopy. b, d, e Astrocytes show clear Ca²⁺ elevations by facial air-puff stimulation (b). Air puffs presented for 10 s induce significant Ca²⁺ increases (d, e; n = 5 sessions, 3 mice). c, f, g Astrocytes do not show detectable cAMP signal changes by air puff (n = 5 sessions, 3 mice). h, i Ca²⁺ activity of LC/NA axons in the cortex was imaged for 3 min (h). The period during air puff was magnified (i), which shows a clear MP signal. j Duration of multipeak signals between before air puff (0–60 s) and during air puff (60–70 s) (n = 4 sessions, 4 mice). Scale bars 20 μm. Analysis was performed from average of individual sessions in e and g and from individual MP signals in j. Box plots: box range, 25–50–75% quatile; square, mean; whiskers, max–min. paired t-test: e; Student’s t-test with Welch’s correction: j; *p < 0.05.