Effect of various kinase inhibitors on the motility of intact sea urchin spermatozoa
| Kinase or pathway targeted . | Substance . | IC50 (μmol l–1) . | Type of action or target . | Concentration range tested . | Inhibition of motility . |
|---|---|---|---|---|---|
| AKT | AKT inhibitor | 5 | Phosphatidyl-inositol ester analog | 1–10 μmol l–1 | None |
| ERK pathway | CGP85793 | ND | Grb2-SH2 domain competitor | 1–100 μmol l–1 | None |
| CGP78850 | ND | Grb2-SH2 domain competitor | 1–100 μmol l–1 | Partial at 100 μmol l–1 | |
| PKA | H-89 | 0.048 | ATP-site competitor | 1–100 μmol l–1 | Total at ⩾30 μmol l–1 |
| KT 5720 | 0.056 | ATP-site competitor | 0.1–10 μmol l–1 | None | |
| dbcAMP | NA | Activator | 1–10 mmol l–1 | None | |
| IBMX | NA | Activator | 0.1–1 mmol l–1 | None | |
| Rp-cAMPS | 11 | cAMP competitor | 0.2–2 mmol l–1 | None | |
| PKC | Chelerythrine | 0.66 | Inhibits PKC translocation | 0.5–50 μmol l–1 | Total at ⩾2 μmol l–1 |
| Calphostin C | 0.05 | Diacylglycerol competitor | 0.15–15 μmol l–1 | Total at ⩾10 μmol l–1 | |
| Gö6976 | 0.002–0.006 | Ca2+ dependent PKC α1 and β1 | 0.05–50 μmol l–1 | Partial at ⩾0.5 μmol l–1 | |
| PI3K | Wortmannin | 0.005 | Catalytic subunit | 1–100 nmol l–1 | None |
| PTK | Tyrphostin A47 | 2.4 | Receptor type PTK | 1–50 μmol l–1 | Partial at ⩾10 μmol l–1 |
| Tyrphostin A1 | >100 | Inactive analog | 0.1–100 μmol l–1 | None | |
| PP2 | 0.005 | Non-receptor type PTK | 0.2–2 μmol l–1 | None | |
| Herbymicin A | 8–12 | Non-receptor type PTK | 10 μmol l–1 | None | |
| Genistein | 2–6 | ATP-site competitor | 100 μmol l–1 | None | |
| poly-Glu-Tyr | NA | Competitor for Tyr phosphorylation | 10 μg ml–1 | None | |
| Broad spectrum | Staurosporine | 0.001–0.03 | ATP-site competitor | 0.2–2 μmol l–1 | Total at ⩾1 μmol l–1 |
| H-7 | 3–6 | ATP-site competitor | 10–100 μmol l–1 | None |
| Kinase or pathway targeted . | Substance . | IC50 (μmol l–1) . | Type of action or target . | Concentration range tested . | Inhibition of motility . |
|---|---|---|---|---|---|
| AKT | AKT inhibitor | 5 | Phosphatidyl-inositol ester analog | 1–10 μmol l–1 | None |
| ERK pathway | CGP85793 | ND | Grb2-SH2 domain competitor | 1–100 μmol l–1 | None |
| CGP78850 | ND | Grb2-SH2 domain competitor | 1–100 μmol l–1 | Partial at 100 μmol l–1 | |
| PKA | H-89 | 0.048 | ATP-site competitor | 1–100 μmol l–1 | Total at ⩾30 μmol l–1 |
| KT 5720 | 0.056 | ATP-site competitor | 0.1–10 μmol l–1 | None | |
| dbcAMP | NA | Activator | 1–10 mmol l–1 | None | |
| IBMX | NA | Activator | 0.1–1 mmol l–1 | None | |
| Rp-cAMPS | 11 | cAMP competitor | 0.2–2 mmol l–1 | None | |
| PKC | Chelerythrine | 0.66 | Inhibits PKC translocation | 0.5–50 μmol l–1 | Total at ⩾2 μmol l–1 |
| Calphostin C | 0.05 | Diacylglycerol competitor | 0.15–15 μmol l–1 | Total at ⩾10 μmol l–1 | |
| Gö6976 | 0.002–0.006 | Ca2+ dependent PKC α1 and β1 | 0.05–50 μmol l–1 | Partial at ⩾0.5 μmol l–1 | |
| PI3K | Wortmannin | 0.005 | Catalytic subunit | 1–100 nmol l–1 | None |
| PTK | Tyrphostin A47 | 2.4 | Receptor type PTK | 1–50 μmol l–1 | Partial at ⩾10 μmol l–1 |
| Tyrphostin A1 | >100 | Inactive analog | 0.1–100 μmol l–1 | None | |
| PP2 | 0.005 | Non-receptor type PTK | 0.2–2 μmol l–1 | None | |
| Herbymicin A | 8–12 | Non-receptor type PTK | 10 μmol l–1 | None | |
| Genistein | 2–6 | ATP-site competitor | 100 μmol l–1 | None | |
| poly-Glu-Tyr | NA | Competitor for Tyr phosphorylation | 10 μg ml–1 | None | |
| Broad spectrum | Staurosporine | 0.001–0.03 | ATP-site competitor | 0.2–2 μmol l–1 | Total at ⩾1 μmol l–1 |
| H-7 | 3–6 | ATP-site competitor | 10–100 μmol l–1 | None |
Immotile concentrated spermatozoa were first diluted in IM in the absence or presence of kinase inhibitor and then in ASW with or without the inhibitor at the same concentration. Motility was assessed by light microscopy with a dark-field illumination as detailed in the Materials and methods, and evaluations of motility inhibition reported here are for the end of the 15-min observation period. ND, not determined; NA, not applicable