Compared to glyt1 siblings, glyt1−/− mutants show delayed activation of pre-motor, and motor regions during emergence from propofol. Four batches of sixteen to eighteen, six-day-old larvae were placed in baskets (diagramed to the left) to enable easy transfer of animals from 10 µM propofol to system water solutions. Half contained glyt1 siblings after (A) ten minutes and (B) twenty minutes in system water. The other half contained glyt1−/− mutants after (C) ten minutes and (D) twenty minutes in system water. To compare activity between propofol and wash treatments, voxels with more intense pERK/tERK at a p < 0.00005 threshold under propofol conditions are shown in purple while voxels with more intense pERK/tERK in system water during emergence are shown in green for glyt1 siblings (A, B) and glyt1−/− mutants in (C, D). glyt1 siblings show greater activation of preoptic, subpallium, and spinal cord regions under propofol, with the olfactory bulb and a regions of the diencephalon that encompasses the arousal pathways (yellow arrowhead in A) being the first brain region to become more active during emergence from propofol, followed ten minutes later by the optic tectum, cerebellum and hindbrain. glyt1−/− mutants have greater activation of preoptic and subpallium, but also optic tectum neuropil and cerebellar neuropil under propofol. As with their glyt1 siblings, the olfactory bulb is the first brain region to become activated but there is no activation of other sensory, pre-motor, and motor regions by the twenty-minute timepoint consistent with delays seen in glyt1−/− mutants recovery of locomotor behaviors after propofol. Scale bars = 100 µm.
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