methiothepin mesylate  (TargetMol)

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: A) Example v-PN cell clusters demonstrating degree of overlap of expression the 5-HT2B LexA with A) the 5-HT2B-T2A-Gal4, B) the 5-HT1A-T2A-Gal4 and C) the 5-HT7-T2A-Gal4. D) Percent of overlap between v-PNs expressed in either the 5-HT2B-LexA (blue) or 5-HT7-LexA (pink) with v-PNs in the T2A-Gal4 lines for the other 5-HT receptors.

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques: Expressing

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: (A) Volumetric reconstructions of v-PNs (green) and lv-PNs (blue) from the Hemibrain EM dataset highlighting the different tracts (arrowheads) formed by the primary processes of each cell cluster. (B) The fascicle formed by the cholinergic v-PNs is highlighted by a GAD1-T2A-LexA driver (white). Neuropil delineated by N-cadherin (blue). (C) The fascicle formed by the cholinergic lv-PNs is highlighted by a ChAT-T2A-LexA driver (white). Neuropil delineated by N-cadherin (blue). (D-U) Example images of intersection between 5-HT receptor T2A-Gal4 (magenta) and LexA (green) drivers. (D-H) Example images from v-PNs of intersection between the 5-HT2B-T2A-LexA and (D) 5-HT1A-T2A-Gal4 (15 ALs from 9 brains), (E) 5-HT1B-T2A-Gal4 (14 ALs from 8 brains), (F) 5-HT2A-T2A-Gal4 (17 ALs from 12 brains), (G) 5-HT2B-T2A-Gal4 (14 ALs from 7 brains) and (H) 5-HT7-T2A-Gal4 (18 ALs from 9 brains). (I-M) Example images from lv-PNs of intersection between the 5-HT2B-T2A-LexA (Average cell body count = 10.97±1.04, n = 73 AL) and (I) 5-HT1A-T2A-Gal4 (12 ALs from 6 brains), (J) 5-HT1B-T2A-Gal4 (16 ALs from 8 brains), (K) 5-HT2A-T2A-Gal4 (17 ALs from 12 brains), (L) 5-HT2B-T2A-Gal4 (13 ALs from 7 brains) and (M) 5-HT7-T2A-Gal4 (17 ALs from 9 brains). (N-Q) Example images from v-PNs of intersection between the 5-HT7-T2A-LexA and (N) 5-HT1A-T2A-Gal4 (19 ALs from 10 brains), (O) 5-HT1B-T2A-Gal4 (16 ALs from 8 brains), (P) 5-HT2A-T2A-Gal4 (16 ALs from 22 brains) and (Q) 5-HT7-T2A-Gal4 (13 ALs from 7 brains). (R-U) Example images from lv-PNs of intersection between the 5-HT7-T2A-LexA (Average cell body count = 5.97±0.48, n = 50 ALs) and (R) 5-HT1A-T2A-Gal4 (12 ALs from 9 brains), (S) 5-HT1B-T2A-Gal4 (12 ALs from 8 brains), (T) 5-HT2A-T2A-Gal4 (15 ALs from 11 brains) and (U) 5-HT7-T2A-Gal4 (11 ALs from 7 brains). All scale bars are 20um.

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques:

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: A) Example image of v-PN morphology within the AL (left panel) and LH (right panel) revealed by MCFO technique using the A) 5-HT1A-T2A-Gal4 (lv-PN), B) 5-HT2A-T2A-Gal4 (lv-PN) and C) 5-HT7-T2A-Gal4 (v-PN). D) Glomerular innervation patterns of v-PNs stochastically labeled via MCFO using each of the 5-HT receptor T2A-Gal4 lines. Each row represents the glomeruli innervated by v-PNs in a single AL with yellow indicating innervation of a given glomerulus and dark blue indicating no innervation. E) Glomerular maps depicting percent of samples in which a given glomerulus was innervated by v-PNs stochastically labeled using each of the 5-HT receptor T2A-Gal4 lines.

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques: Labeling

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: (A) Hierarchical clustering analysis of the glomerular innervation patterns of glomeruli innervated by v-PNs and lv-PNs stochastically labeled via MCFO for each 5-HT receptor T2A-Gal4 line. A total of nine clusters (I-IX, each with their own color) were obtained and clones within each cluster are colored based on the MiMIC T2A-Gal4 from which they derive (green; 5-HT1A, yellow; 5-HT1B, salmon; 5-HT2A, pink; 5-HT2B, cyan; 5-HT7).

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques: Labeling, Clone Assay

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: A) v-PNs expressing different serotonin receptors project to distinct regions of the olfactory system. A) Morphology of the v-PNs within the R24H08-Gal4 (green) within the AL and LH. Immunolabeling for N-Cadherin (blue) highlights neuropil. B) Morphology of the v-PNs within the R86G06-Gal4 (cyan) within the AL and LH. C) Map of glomeruli innervated by R24H08 v-PNs. D) Map of glomeruli innervated by R86G06 v-PNs. E) R24H08 v-PNs overlap with the 5-HT1A-T2A-Gal4. F) R24H08 v-PNs overlap with the 5-HT1B-T2A-Gal4. G) R86G06 v-PNs overlap with the 5-HT7-T2A-Gal4. H) Percent of v-PNs within the R24H08 (green) and R86G06 (blue) LexAs expressed within each 5-HT receptor T2A-Gal4 line. I) Reconstruction of the v-PNs from the Hemibrain dataset that most closely resemble the R24H08 (v-PN R24 ; green) and R86G06 (v-PN R86 ; dark blue) driver lines within AL and LH volumes. R24H08 v-PNs are shown in two shades of green to highlight the v-PNs predominantly innervating the VL2a glomerulus (dark green) and the DC3 glomerulus (light green). J) Reconstruction of the axon terminals of the two v-PN groups within the LH. K) overlap of AL (left) and LH (right) projections of the R24H08 (green) and R86G06 (cyan) v-PNs. Immunolabeling for N-Cadherin (blue) highlights neuropil. L) Synapse fractions of the downstream LH neurons targeted by the candidate v-PNs from the R24H08 and R86G06 driver lines. LH output neurons (LHONs; lavendar), LH local neurons (LHLNs; purple), LH centrifugal neurons (LHCENTs; pink). M) Graph plot of the downstream synaptic targets of the v-PN R24 (green) and v-PN R86 (cyan) shows very little convergence, except for a single LHLN that is targeted by two v-PN R24 and all four v-PN R86 (black arrows). All scale bars = 20um.

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques: Expressing, Immunolabeling

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: A) Calcium imaging responses (peak %ΔF/F) of R24H08 (green) and R86G06 (cyan) v-PNs to a panel of odors. AA; acetoin acetate, ACE; acetophenone, ACV; apple cider vinegar, BEZ; benzaldehyde, FAR; farnesol, 1-HEX; 1-hexanol, 1-OCT; 1-octen-3-ol, OrP; orange peel (n=10). B) R24H08 v-PNs respond to farnesol, but not 1-octen-3-ol, while R86G06 v-PNs respond to 1-octen-3-ol, but not farnesol. C) Average responses of R86G06 v-PNs to 1-octen-3-ol or 1-hexanol before (gray traces) and after application of 5-HT (dark blue) or a 5-HT7 receptor agonist (AS-19; cyan). Arrowhead indicates time of odor application. D) 5-HT and AS-19 enhance odor-evoked responses of R86G06 v-PNs. 5-HT, n=10 (1-Oct), p=0.0057; n=7 (1-Hex), p=0.0053; AS-19, n=10 (1-Oct), p=0.0003; n=8 (1-Hex), p=0.0002; 3 repeated stimulations, significance tested using paired samples T-test. E) RNAi knockdown of the 5-HT7 receptor in R86G06 v-PNs results in a loss of 5-HT induced enhancement of 1-hexanol responses and causes AS-19 to suppress R86G06 v-PN responses. 5-HT, n=10 (1-Oct), p=0.0051; n=9 (1-Hex) n.s.; AS-19, n=11 (1-Oct), p=0.0041; n=12 (1-Hex) p=0.0323; 3 repeated stimulations, significance tested using paired sample T-test. All recordings were made from R86G06 v-PN axon terminals in the lateral horn. F) Average responses of R24H08 v-PNs to farnesol (Far) or benzaldehyde (Bez) before (gray traces) and after application of 5-HT (blue) or a 5-HT1A/1B receptor agonist (Ipsapirone; green). Arrowhead indicates time of odor application. G) 5-HT does not significantly affect odor-evoked responses of R24H08 v-PNs, while ipsapirone reduces responses. 5-HT, n= 7 (Far), n.s.; n=11 (Bez), n.s.; Ipsapirone, n=7 (Far), p=0.0016; n=11 (Bez), 3 repeated stimulations, significance tested using paired sample T-test. H) RNAi knockdown of the 5-HT1A receptor in R24H08 v-PNs results in a loss of the ipsapirone induced reduction of odor-evoked responses. 5-HT, n=8 (Far), n.s.; n=9 (Bez), n.s.; Ipsapirone, n=14 (Far), n.s.; n=13 (Bez), n.s.; 3 repeated stimulations, significance tested using paired sample T-test. All recordings were made from R24H08 v-PN processes in the AL.

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques: Imaging, Cider Assay

Journal: bioRxiv

Article Title: Stimulus-Specific Modulation is Enabled by Differential Serotonin Receptor Expression

doi: 10.1101/2023.06.21.546011

Figure Lengend Snippet: (A) Average responses of R86G06 v-PNs to 1-octen-3-ol (1-Oct) or 1-hexanol (1-Hex) before (gray traces) and after application of the pan-5-HT receptor antagonist metitipine (Red) and the the 5-HT7 receptor antagonist SB269970 (purple). (B) Neither metitipine nor SB269970 affected odor-evoked responses of R86G06 v-PNs. Metitipine, n=11 (1-Oct), n=11 (1-Hex); SB269970, n=10 (1-Oct), n=11 (1-Hex) animals, 3 repeats, ns; paired sample t-test. (C) Average responses of R24H08 v-PNs recorded from processes in the lateral horn to farnesol (Far) or benzaldehyde (Bez) before (gray traces) and after application of 5-HT (blue) and the 5-HT1A/1B receptor agonist ipsapirone (green) and the the 5-HT7 receptor antagonist WAY100635 (orange). (D) Neither 5-HT nor ipsapirone affected odor-evoked responses of R24H08 v-PNs recorded within the lateral horn. 5-HT, n= 11 (Far), n=12 (Bez); Ipsapirone, n=8 (Far), n=7 (Bez) animals, 3 repeats, ns; paired sample t-test. (E) Average responses of R24H08 v-PNs recorded from processes in the lateral horn to farnesol (Far) or benzaldehyde (Bez) before (gray traces) and after application of the pan-5-HT receptor antagonist metitipine (Red) and the the 5-HT1A receptor antagonist WAY100635 (orange). (F) Neither metitipine nor WAY100635 affected odor evoked responses of R24H08 v-PNs recorded within the lateral horn. Metitipine, n=6 (Far), n=6 (Bez); WAY100635, n=7 (Far), n=7 (Bez) animals, 3 repeats, ns; paired sample t-test. (G) Average response of R86G06 v-PNs and R24H08 v-PNs to bath application of AS-19 and ipsapirone respectively over 8 minutes. N= 7-12 samples.

Article Snippet: Pharmacological agents and their concentrations used in this study: 5-HT receptor agonist (10 -5 M serotonin hydrochloride, TCI Chemicals, CAS:153-98-0) was made fresh every day before the start of experiments and the aliquot was shielded from light, 5-HT receptor antagonist (10 -5 M methiothepine mesylate salt, Sigma, CAS:74611-28-2), 5-HT1A receptor antagonist (10 -5 M WAY-100635 maleate, Abcam, CAS:634908-75-1), 5-HT1A receptor agonist (10 -5 M Ipsapirone, Rndsystems, CAS:95847-70-4), 5-HT7 receptor antagonist (10 -5 M Sb 258719, Tocris, CAS:1217674-10-6), 5-HT7 receptor agonist (10 -5 M AS-19, Tocris, CAS:1000578-26-6).

Techniques: