Fig. 2

Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) cells exhibit distinct spontaneous activity patterns in vivo. (A) A confocal image of the pituitary shows RCaMP2 expression in both cell types and GFP expression in FSH cells (scale bar = 100 µm). (B) A diagram describing the setup of the in vivo experiments. The dissected zebrafish were placed in a chamber with a constant flow of water to the gills and imaged in an upright two-photon microscopy. (C) A representative image of in vivo calcium activity (see Figure 2?video 3). On the top left is a merged image depicting FSH cells in green and LH cells in magenta. The other top panels show sequential calcium imaging calcium rise in FSH cells is marked by white arrows. The bottom panels show the calcium of LH and FSH cells in two different imaged pituitaries, one where only FSH cells were active (Fish 1) and another where both cell types were active (Fish 2, traces ?F/F, see Figure 2?figure supplement 1A. for heatmap of the calcium traces, scale bar = 20 µm). (D) The properties of spontaneous calcium transients (?F/F) in LH cells and FSH cells in three males and one female and their means. (unpaired t-test, **p<0.001). Analysis was performed using pCLAMP 11. (E) Left: cross-correlation analysis between active ROI to the rest of the cell. The color-coded data points are superimposed on the imaged cells and represent the maximum cross-correlation coefficient between a calcium trace of a region of interest (ROI) and that of the rest of the cells in the same population. Right: is a matrix of maximum cross-correlation coefficient values between all the cells. (scale bar = 20 µm). (F) Summary of the mean max cross-correlation coefficient values of calcium traces in each cell population of repeated in vivo calcium imaging assays (n (calcium sessions)=16, see Figure 2?figure supplement 1B for all measurements,. unpaired t-test, ***p<0.0001).