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. 2022 Sep 26;32(18):3952–3970.e8. doi: 10.1016/j.cub.2022.07.038

Figure 6.

Figure 6

Astrocytes are differentially responsive to neurotransmitters

(A) Illustration of possible glutamate-evoked D-serine release from astrocytes.

(B) Astrocytes adjacent to the pars intercerebralis show differential responses to glutamate application (green bar).

(C) Protocol for drug application with randomized order for acetylcholine (ACh), ATP, and glutamate (Glu).

(D) Average traces for ACh, ATP, and glutamate partitioned by the type of response (activated, no change, or inhibited). Responses were determined as follows: activated if μpost-drug > σpre-drug + μpre-drug, no change if μpost-drug fell within σpre-drug + μpre-drug and inhibited if μpost-drug < σpre-drug − μpre-drug. Line is smoothed average and shaded band is SEM.

(E) Neurotransmitters induce both excitatory and inhibitory responses in astrocytes. Paired datapoints represent responses from a single astrocyte, and group is assembled from multiple flies (nAch = 16, 68, 39; nATP = 86, 55, 149; nglut = 52, 140, 97). ∗∗p < 0.01, Kruskal-Wallis ANOVA with Dunn’s multiple comparisons test.

(F) Proportions of astrocytes sorted by response direction following acetylcholine, ATP, and glutamate application.

(G) Average ΔF/F0 during drug application for glutamate plotted against ΔF/F0 for ATP shows most astrocytes that are excited by glutamate are also excited by ATP and vice versa.

(H) Astrocytes are more likely to have matched responses between glutamate and ATP. Chi-square test between matched versus mismatched astrocytes. Fisher’s exact test matched versus mismatched p = 2.35e−11, OR = 3.24.

(I) Venn diagram showing considerable overlap of matched responses of astrocytes to all three neurotransmitters.

(J) Protocol for tetanus toxin (TetX) application.

(K) Average traces for glutamate- and ATP-evoked excitatory and inhibitory responses with and without TetX. Line is smoothed average and shaded band SEM.

(L) Blocking vesicular transmission does not suppress astrocyte responses to transmitter application (left to right, nPre-TetXATP = 21, 6; nPost-TetXATP = 16, 35; nPre-TetXGlut = 18, 11; nPost-TetXGlut = 14, 9). n.s. > 0.05 Kruskal-Wallis ANOVA with Dunn’s multiple comparisons test. Individual data points are single astrocytes across multiple animals.

See also Figure S6.