5α-cyprinol sulfate, a bile salt from fish, induces diel vertical migration in Daphnia
- University of Koeln, Germany
- University of Zurich, Switzerland
Figures
Figure 1 with 1 supplement
Chromatogram and biological activity of extracted fish incubation water (EFI).
(A) Chromatogram of EFI after separation on a 250 mm x 4 mm reversed phase column (Nucleosil 100–5 C18, Macherey-Nagel, Düren, Germany) using the ammonium acetate buffered mobile phases A H2O and B …
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Figure 1—source data 1
Temperature profile of the experimental tubes.
- https://doi.org/10.7554/eLife.44791.004
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Figure 1—source data 2
Response of Daphnia to HPLC fractions.
Statistical analysis of mean daytime residence depth of Daphnia magna in response to different HPLC-fractions (Frac 1 - Frac 6) of extracted fish incubation water (EFI) as shown in Figure 1B. Significantly different pairwise comparisons are given in red, n.s.: not significant.
- https://doi.org/10.7554/eLife.44791.005
Figure 1—figure supplement 1
Temperature profile of the experimental tubes.
https://doi.org/10.7554/eLife.44791.003
Figure 2
MS data of the biologically active HPLC fraction.
(A) ESI-MS spectrum extracted from the time window corresponding to the active fraction three in Figure 1 (11 - 14 min) measured in the negative ionization mode. m/z 531.29986 [M–H]– indicates the …
Figure 3 with 2 supplements
Retention times of selected bile salts in extracted fish incubation water and fish bile.
Extracted ion chromatograms of cyprinol sulfate ([M]–calc=m/z 532.3070; black line) and the conjugated bile acid taurochenodeoxycholic acid (TCDCA) ([M]–calc=m/z 498.2889; dashed line) after ESI-MS …
Figure 3—figure supplement 1
MS-MS (negative ionization mode) of 531.2997 [M-H]-: 96.95980–96.95992 at 50 eV for (A) 5α-CPS purified from carp bile, (B) extracted fish incubation water (EFI) of R.rutilus and (C) extracted bile of rainbow trout Oncorhynchus mykiss for the time windows, when α-CPS (Mcalc = m/z 531.29986) was detected during LC-MS of Figure 3.
https://doi.org/10.7554/eLife.44791.008-
Figure 3—figure supplement 1—source data 1
MS-MS measurements of m/z=531.2997 deriving from fish incubation water and fish bile.
- https://doi.org/10.7554/eLife.44791.009
Figure 3—figure supplement 2
Behavioral response of Daphnia magna to extract of fish incubation water (EFI) and to extract of carp and trout bile.
Controls contain equivalent volumes of pure organic solvent. Mean daytime residence depth (± SD, n = 4). The day:night cycle was 16:8 hr. Different capital letters indicate significant differences …
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Figure 3—figure supplement 2—source data 1
Response of Daphnia to extracts of fish incubation water and fish bile.
Statistical analysis of mean daytime residence depth of Daphnia magna in response to extracted fish incubation water (EFI) and to extract of carp bile and trout bile as shown in Figure 3—figure supplement 2. Significantly different pairwise comparisons are given in red, n.s.: not significant.
- https://doi.org/10.7554/eLife.44791.011
Figure 4 with 3 supplements
Effects of extracted fish incubation water (EFI) and of increasing concentrations of selected bile salts.
(A) 5α-cyprinol sulfate (5α-CPS), (B) taurochenodeoxycholic acid (TCDCA), and. (C) chenodeoxycholic acid (CDCA) on behavioral responses of Daphnia. Controls contain equivalent volumes of pure …
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Figure 4—source data 1
Effects of extracted fish incubation water (EFI) and of increasing concentrations of selected bile salts on diel vertical migration.
- https://doi.org/10.7554/eLife.44791.017
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Figure 4—source data 2
Response of Daphnia to 5α-cyprinol sulfate.
Statistical analysis of mean daytime residence depth of Daphnia magna in response to different concentrations of 5α-cyprinol sulfate (5α-CPS) as shown in Figure 4A. Significantly different pairwise comparisons are given in red, n.s.: not significant.
- https://doi.org/10.7554/eLife.44791.018
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Figure 4—source data 3
Response of Daphnia to 5α-cyprinol sulfate.
Statistical analysis of mean daytime residence depth of Daphnia magna in response to different concentrations of taurochenodeoxycholic acid (TCDCA) as shown in Figure 4B. Significantly different pairwise comparisons are given in red, n.s.: not significant.
- https://doi.org/10.7554/eLife.44791.019
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Figure 4—source data 4
Response of Daphnia to chenodeoxycholic acid (CDCA).
Statistical analysis of mean daytime residence depth of Daphnia magna in response to different concentrations of chenodeoxycholic acid (CDCA) as shown in Figure 4C). Significantly different pairwise comparisons are given in red, n.s.: not significant.
- https://doi.org/10.7554/eLife.44791.020
Figure 4—figure supplement 1
Total ion chromatogram of 5α-cyprinol sulfate (α-CPS) purified from fish bile (Cyprinus carpio).
Stacked total ion chromatograms of purified α-CPS in MeOH/MilliQ (blue line) and a blank run (1:1 MeOH/MilliQ, red line) after ESI-MS in different ionization modes. Total ion counts are depicted. (A)…
Figure 4—figure supplement 2
MS data of the subfraction from 1 to 3 min of LC-MS runs of purified 5α-cyprinol sulfate (α-CPS) and a blank sample (SI Figure 4).
ESI-MS spectra extracted from the time window of 1–3 min in SI Figure 4. Obtained spectra for α-CPS (A, B) and blank measurements (MeOH/MilliQ, 1:1, v:/; C,D) measured either after negative or …
Figure 4—figure supplement 3
Calibration curves after ESI-MS of (A) taurochenodeoxycholate (TCDCA) and (B) cholesteryl sulfate used for the quantification of 5α-cyprinol sulfate.
The detected peak areas (PA) of cholesteryl sulfate and TCDCA normalized to the detected peak areas of glycocholic acid (internal standard) are plotted against the respective concentrations.
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Figure 4—figure supplement 3—source data 1
Calibration curves of taurochenodeoxycholate and cholesteryl sulfate.
- https://doi.org/10.7554/eLife.44791.016
Author response image 1
Chromatogram and biological activity of extracted fish incubation water (EFI).
(a) Chromatogram of EFI after separation on a 250 mm x 4 mm reversed phase column (Nucleosil 100-5-C18, Macherey-Nagel, Düren, Germany) using the mobile phases A H2O and B MeOH with the portion of B …
Author response image 2
ESI-MS data of the HPLC fractions leading to daytime depths of Daphnia magna which do neither statistically differ from the negative nor from the postive control and the corresponding spectra of a blank run.
ESI-MS spectrum extracted from the time window corresponding to fraction: Author response image 2“Pre”, Author response image 4“8”, Author response image 6“9” in Author response image 1 and Author …
Author response image 3
Author response image 4
Author response image 5
Author response image 6
Author response image 7
Author response image 8
MS/MS of m/z =531.3 in fraction “8”.
Negative ionization MS/MS of m/z 531 [M– H]–:96.95979 (HSO4–calc=m/z 96.96010, ∆ = –3.2ppm), which indicated the presence of a sulfate group in m/z 531.30048 [M– H]–.
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.44791.021
