Fig 1.
Calculation of Learning and Memory Indices.
For all experiments, a courtship index (CI) was calculated as the percent of time spent performing courtship behaviors in three different time periods: the ten minutes at the beginning of training (CIbegin), the ten minutes at the end of training (CIend), and the ten minute test period (CItest). In this example, the CIs for each of these three periods are plotted for wild-type DL flies tested at 25C. To generate the learning index (LI), we subtract CIbegin−CIend. A significantly positive LI indicates that learning occurred. To generate the memory index (MI), we subtract CIend−CItest. An MI close to zero indicates that memory occurred, and a significantly negative MI indicates a lack of memory. In these flies, significant learning occurred, but there was not a significant change between the CIend and the CItest, indicating memory also occurred. Significance is determined using a one-sided Wilcoxon signed rank test; learning significance is a comparison between CIbegin and CIend, and memory significance is a comparison between CIend and CItest. ****, p < .0001. Error bars are SEM, n = 48.
Fig 2.
Courtship Conditioning Learning and Memory in Kenyon Cell Lines.
Learning index (LI) and memory index (MI) for A. α/β Kenyon cell lines, B. secondary α/β Kenyon cell lines, and C. α’/β’ Kenyon cell lines. Lines identified as courtship memory hits are boxed in red. Expression patterns are directly below the LI and MI for each line. Shading indicates relative levels of expression in each neuron type as reported in [35]. Significance is determined using one-sided Wilcoxon signed ranks tests with Benjamini-Hochberg corrections on primary screen data. *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. Error bars are SEM, n = 19–46. D. Diagrams of the mushroom bodies and the general area of expression of the α/βc neurons [35].
Fig 3.
Courtship Conditioning Learning and Memory in MB Output Neurons.
Learning index (LI) and memory index (MI) for A. xγ2α’1 and α’2 MBON lines, B. γ2β’2a and β’2mp MBON lines, C. γ3 and γ3β’1 MBON lines, and D. α1 MBON line. Lines identified as courtship memory hits are boxed in red. Expression patterns are directly below the LI and MI for each line. Shading indicates relative levels of expression as reported in [35]. Significance is determined using one-sided Wilcoxon signed rank tests with Benjamini-Hochberg post-hoc corrections. *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. Error bars are SEM, n = 9–24. Secondary screening for E. γ2α‘1 and α‘2 MBON lines, F.γ2β’2a and β’2mp MBON lines, G. γ3 and γ3β’1 MBON lines, and H. α1 MBON lines. Lines identified as courtship memory hits are boxed in red. Expression patterns are directly below the LI and MI for each line. Shading indicates relative levels of expression as reported in [35]. Significance is determined using one-sided Wilcoxon signed rank tests. *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. Error bars are SEM, n = 8–46. I. Diagram of the mushroom bodies and the sites of innervation of the γ2α‘1 and α‘2 MBONs, γ3 and γ3β’1 MBONs, γ5β’2a and β’2mp MBONs, and α1 MBONs [35].
Fig 4.
Courtship Conditioning Learning and Memory in PAM Neurons.
Learning index (LI) and memory index (MI) for A. broadly-expressed PAM neuron lines, B. PAM γ lines, and C. PAM α1 lines. Lines identified as courtship memory hits are boxed in red. Expression patterns are directly below the LI and MI for each line. Shading indicates relative levels of expression as reported in [35]. Significance is determined using one-sided Wilcoxon signed rank tests with Benjamini-Hochberg post-hoc corrections. *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. Error bars are SEM, n = 19–24. Secondary screening for D. PAM γ lines and E. PAM α1 lines. Lines identified as courtship memory hits are boxed in red. Expression patterns are directly below the LI and MI for each line. Shading indicates relative levels of expression as reported in [35]. Significance is determined using one-sided Wilcoxon signed rank tests. *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. Error bars are SEM, n = 17–27. F. Diagram of the mushroom bodies, the general location of the cluster of PAM neuron cell bodies, and the sites of innervation of the γ3, γ5, and α1, β1, β’1 PAM neuron lines [35].