Journal: bioRxiv

Article Title: Rapid sensory integration in orexin neurons governs probability of future movements

doi: 10.1101/620096

Figure Lengend Snippet: A, A virus was used to drive GCaMP6s expression under the orexin promoter, such that only orexin cells express GCaMP6s. B, An orexin neuron in a brain slice was imaged with a miniature endoscope at an angle (above micrographs: AVG, average time projection; STD, standard deviation projection; recorded neuron at arrow head) and with patch clamp microscope from above (below left, GCaMP6s fluorescence; below right, merged GCaMP6s, Alexa-568 red patch pipette filling solution and oblique illumination micrograph showing patch pipette; recorded neuron at arrow head). C, Ca-imaging (green, Z-scored ΔF/F 0 ), electrophysiology (black) and firing rate (red) example data from cell in B. D, Expanded view of firing burst onset showing short latency from firing burst to Ca increase. E, Combined data across 6 recordings (each recording in a different colour) showing linear relation between maximal fluorescence and firing rate during each 10s current step. R 2 of linear fit is 0.91. F, Schematic of freely-moving in vivo paradigm where orexin neurons in subject (s) were recorded with miniature endoscope (m) in an arena with a conspecific (c) and food pod (f) while behaviour was tracked from below through the transparent floor. G, Representative micrographs showing orexin immuno-reactivity (IR) and GCaMP6s in coronal sections prepared from an experimental subject. GRIN lens track is visible dorsal to the orexin neurons. H, Representative behaviour video frames showing a female subject (black fur, blue dashed outline) feeding (left), mating (second from left), 3 different medium speed movements (second from right) and being chased (right) by a male conspecific (brown fur, white dashed outline). I, Representative average time projection of Ca-imaging data through the cylindrical GRIN lens. J, Representative time series of z-scored fluorescence (ΔF/F 0 ) of 28 orexin neurons across various behavioural epochs. K, Histogram of Pearson correlation coefficients across all recorded cells (mean correlation with bout speed 0.43 ± 0.02; with distance 0.27 ± 0.03; with duration −0.14 ± 0.02; each distribution significantly different from the others, P<10 -17 with paired Student’s t-test; 207 cells across 12 recording sessions from 5 animals, 2 male, 3 female). L, Mean Ca-activity for each cell during each behaviour bout (points) and their average (squares) coloured by the behaviour class (colour code from red to green follows roughly movement speed in behaviour) plotted as a function of mean locomotion speed in behaviour bout (249 bouts, 5873 cell/bout pairs). Black line is linear fit to bout average data (squares) with R 2 =0.21 and P<10 -13 . M and N, Same data as in L grouped into behaviour classes and plotted as mean ± s.d.

Article Snippet: For imaging experiments, the optical device was a 0.39 NA, 7.3 mm long, 0.6 mm diameter cylindrical graded refractive index (GRIN) lens (Grintech).

Techniques: Expressing, Slice Preparation, Standard Deviation, Patch Clamp, Microscopy, Fluorescence, Transferring, Imaging, In Vivo, Activity Assay