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. 1999 Jun 1;340(Pt 2):365–369.

Signal transduction pathway regulating prostaglandin EP3 receptor-induced neurite retraction: requirement for two different tyrosine kinases.

J Aoki 1, H Katoh 1, H Yasui 1, Y Yamaguchi 1, K Nakamura 1, H Hasegawa 1, A Ichikawa 1, M Negishi 1
PMCID: PMC1220258  PMID: 10333476

Abstract

We reported previously that activation of the prostaglandin E receptor EP3 subtype triggered neurite retraction through the small GTPase Rho-, and its target, RhoA-binding kinase alpha (ROKalpha)-, dependent pathway in EP3 receptor-expressing PC12 cells. Here we examined the involvement of tyrosine kinases in this pathway in nerve growth factor-differentiated PC12 cells. Tyrphostin A25, a tyrosine kinase inhibitor, blocked neurite retraction and cell rounding induced by activation of the EP3 receptor, however, it failed to block neurite retraction and cell rounding induced by microinjection of constitutively active RhoA, RhoAV14, indicating that a tyrphostin-sensitive tyrosine kinase was involved in the pathway from the EP3 receptor to Rho activation. On the other hand, genistein, another tyrosine kinase inhibitor, blocked neurite retraction and cell rounding induced by both activation of the EP3 receptor and microinjection of RhoAV14. However, genistein did not block neuronal morphological changes induced by microinjection of a constitutively active mutant of ROKalpha. These results indicate that two different tyrosine kinases, tyrphostin A25-sensitive and genistein-sensitive kinases, are involved in the EP3 receptor-mediated neurite retraction acting upstream and downstream of Rho, respectively.

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Selected References

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  1. Aktories K., Weller U., Chhatwal G. S. Clostridium botulinum type C produces a novel ADP-ribosyltransferase distinct from botulinum C2 toxin. FEBS Lett. 1987 Feb 9;212(1):109–113. doi: 10.1016/0014-5793(87)81566-1. [DOI] [PubMed] [Google Scholar]
  2. Amano M., Chihara K., Kimura K., Fukata Y., Nakamura N., Matsuura Y., Kaibuchi K. Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase. Science. 1997 Feb 28;275(5304):1308–1311. doi: 10.1126/science.275.5304.1308. [DOI] [PubMed] [Google Scholar]
  3. Amano M., Chihara K., Nakamura N., Fukata Y., Yano T., Shibata M., Ikebe M., Kaibuchi K. Myosin II activation promotes neurite retraction during the action of Rho and Rho-kinase. Genes Cells. 1998 Mar;3(3):177–188. doi: 10.1046/j.1365-2443.1998.00181.x. [DOI] [PubMed] [Google Scholar]
  4. Amano M., Ito M., Kimura K., Fukata Y., Chihara K., Nakano T., Matsuura Y., Kaibuchi K. Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase). J Biol Chem. 1996 Aug 23;271(34):20246–20249. doi: 10.1074/jbc.271.34.20246. [DOI] [PubMed] [Google Scholar]
  5. Feng J., Ito M., Kureishi Y., Ichikawa K., Amano M., Isaka N., Okawa K., Iwamatsu A., Kaibuchi K., Hartshorne D. J. Rho-associated kinase of chicken gizzard smooth muscle. J Biol Chem. 1999 Feb 5;274(6):3744–3752. doi: 10.1074/jbc.274.6.3744. [DOI] [PubMed] [Google Scholar]
  6. Ferreira S. H. Prostaglandins, aspirin-like drugs and analgesia. Nat New Biol. 1972 Dec 13;240(102):200–203. doi: 10.1038/newbio240200a0. [DOI] [PubMed] [Google Scholar]
  7. Flinn H. M., Ridley A. J. Rho stimulates tyrosine phosphorylation of focal adhesion kinase, p130 and paxillin. J Cell Sci. 1996 May;109(Pt 5):1133–1141. doi: 10.1242/jcs.109.5.1133. [DOI] [PubMed] [Google Scholar]
  8. Hall A. Rho GTPases and the actin cytoskeleton. Science. 1998 Jan 23;279(5350):509–514. doi: 10.1126/science.279.5350.509. [DOI] [PubMed] [Google Scholar]
  9. Hirose M., Ishizaki T., Watanabe N., Uehata M., Kranenburg O., Moolenaar W. H., Matsumura F., Maekawa M., Bito H., Narumiya S. Molecular dissection of the Rho-associated protein kinase (p160ROCK)-regulated neurite remodeling in neuroblastoma N1E-115 cells. J Cell Biol. 1998 Jun 29;141(7):1625–1636. doi: 10.1083/jcb.141.7.1625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jalink K., Eichholtz T., Postma F. R., van Corven E. J., Moolenaar W. H. Lysophosphatidic acid induces neuronal shape changes via a novel, receptor-mediated signaling pathway: similarity to thrombin action. Cell Growth Differ. 1993 Apr;4(4):247–255. [PubMed] [Google Scholar]
  11. Jalink K., Moolenaar W. H. Thrombin receptor activation causes rapid neural cell rounding and neurite retraction independent of classic second messengers. J Cell Biol. 1992 Jul;118(2):411–419. doi: 10.1083/jcb.118.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jalink K., van Corven E. J., Hengeveld T., Morii N., Narumiya S., Moolenaar W. H. Inhibition of lysophosphatidate- and thrombin-induced neurite retraction and neuronal cell rounding by ADP ribosylation of the small GTP-binding protein Rho. J Cell Biol. 1994 Aug;126(3):801–810. doi: 10.1083/jcb.126.3.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Katoh H., Aoki J., Ichikawa A., Negishi M. p160 RhoA-binding kinase ROKalpha induces neurite retraction. J Biol Chem. 1998 Jan 30;273(5):2489–2492. doi: 10.1074/jbc.273.5.2489. [DOI] [PubMed] [Google Scholar]
  14. Katoh H., Aoki J., Yamaguchi Y., Kitano Y., Ichikawa A., Negishi M. Constitutively active Galpha12, Galpha13, and Galphaq induce Rho-dependent neurite retraction through different signaling pathways. J Biol Chem. 1998 Oct 30;273(44):28700–28707. doi: 10.1074/jbc.273.44.28700. [DOI] [PubMed] [Google Scholar]
  15. Katoh H., Negishi M., Ichikawa A. Prostaglandin E receptor EP3 subtype induces neurite retraction via small GTPase Rho. J Biol Chem. 1996 Nov 22;271(47):29780–29784. doi: 10.1074/jbc.271.47.29780. [DOI] [PubMed] [Google Scholar]
  16. Kimura K., Ito M., Amano M., Chihara K., Fukata Y., Nakafuku M., Yamamori B., Feng J., Nakano T., Okawa K. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase) Science. 1996 Jul 12;273(5272):245–248. doi: 10.1126/science.273.5272.245. [DOI] [PubMed] [Google Scholar]
  17. Kozma R., Sarner S., Ahmed S., Lim L. Rho family GTPases and neuronal growth cone remodelling: relationship between increased complexity induced by Cdc42Hs, Rac1, and acetylcholine and collapse induced by RhoA and lysophosphatidic acid. Mol Cell Biol. 1997 Mar;17(3):1201–1211. doi: 10.1128/mcb.17.3.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leung T., Chen X. Q., Manser E., Lim L. The p160 RhoA-binding kinase ROK alpha is a member of a kinase family and is involved in the reorganization of the cytoskeleton. Mol Cell Biol. 1996 Oct;16(10):5313–5327. doi: 10.1128/mcb.16.10.5313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Namba T., Sugimoto Y., Negishi M., Irie A., Ushikubi F., Kakizuka A., Ito S., Ichikawa A., Narumiya S. Alternative splicing of C-terminal tail of prostaglandin E receptor subtype EP3 determines G-protein specificity. Nature. 1993 Sep 9;365(6442):166–170. doi: 10.1038/365166a0. [DOI] [PubMed] [Google Scholar]
  20. Needham L. K., Rozengurt E. Galpha12 and Galpha13 stimulate Rho-dependent tyrosine phosphorylation of focal adhesion kinase, paxillin, and p130 Crk-associated substrate. J Biol Chem. 1998 Jun 5;273(23):14626–14632. doi: 10.1074/jbc.273.23.14626. [DOI] [PubMed] [Google Scholar]
  21. Negishi M., Sugimoto Y., Ichikawa A. Molecular mechanisms of diverse actions of prostanoid receptors. Biochim Biophys Acta. 1995 Oct 26;1259(1):109–119. doi: 10.1016/0005-2760(95)00146-4. [DOI] [PubMed] [Google Scholar]
  22. Negishi M., Sugimoto Y., Ichikawa A. Prostaglandin E receptors. J Lipid Mediat Cell Signal. 1995 Oct;12(2-3):379–391. doi: 10.1016/0929-7855(95)00024-k. [DOI] [PubMed] [Google Scholar]
  23. Postma F. R., Jalink K., Hengeveld T., Moolenaar W. H. Sphingosine-1-phosphate rapidly induces Rho-dependent neurite retraction: action through a specific cell surface receptor. EMBO J. 1996 May 15;15(10):2388–2392. [PMC free article] [PubMed] [Google Scholar]
  24. Reimann W., Steinhauer H. B., Hedler L., Starke K., Hertting G. Effect of prostaglandins D2, E2 and F2alpha on catecholamine release from slices of rat and rabbit brain. Eur J Pharmacol. 1981 Feb 19;69(4):421–427. doi: 10.1016/0014-2999(81)90445-3. [DOI] [PubMed] [Google Scholar]
  25. Ridley A. J., Hall A. Signal transduction pathways regulating Rho-mediated stress fibre formation: requirement for a tyrosine kinase. EMBO J. 1994 Jun 1;13(11):2600–2610. doi: 10.1002/j.1460-2075.1994.tb06550.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sekine A., Fujiwara M., Narumiya S. Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase. J Biol Chem. 1989 May 25;264(15):8602–8605. [PubMed] [Google Scholar]
  27. Sugimoto Y., Shigemoto R., Namba T., Negishi M., Mizuno N., Narumiya S., Ichikawa A. Distribution of the messenger RNA for the prostaglandin E receptor subtype EP3 in the mouse nervous system. Neuroscience. 1994 Oct;62(3):919–928. doi: 10.1016/0306-4522(94)90483-9. [DOI] [PubMed] [Google Scholar]
  28. Suidan H. S., Stone S. R., Hemmings B. A., Monard D. Thrombin causes neurite retraction in neuronal cells through activation of cell surface receptors. Neuron. 1992 Feb;8(2):363–375. doi: 10.1016/0896-6273(92)90302-t. [DOI] [PubMed] [Google Scholar]
  29. Tigyi G., Fischer D. J., Sebök A., Yang C., Dyer D. L., Miledi R. Lysophosphatidic acid-induced neurite retraction in PC12 cells: control by phosphoinositide-Ca2+ signaling and Rho. J Neurochem. 1996 Feb;66(2):537–548. doi: 10.1046/j.1471-4159.1996.66020537.x. [DOI] [PubMed] [Google Scholar]
  30. Tigyi G., Miledi R. Lysophosphatidates bound to serum albumin activate membrane currents in Xenopus oocytes and neurite retraction in PC12 pheochromocytoma cells. J Biol Chem. 1992 Oct 25;267(30):21360–21367. [PubMed] [Google Scholar]
  31. Uehata M., Ishizaki T., Satoh H., Ono T., Kawahara T., Morishita T., Tamakawa H., Yamagami K., Inui J., Maekawa M. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature. 1997 Oct 30;389(6654):990–994. doi: 10.1038/40187. [DOI] [PubMed] [Google Scholar]
  32. Wolfe L. S. Eicosanoids: prostaglandins, thromboxanes, leukotrienes, and other derivatives of carbon-20 unsaturated fatty acids. J Neurochem. 1982 Jan;38(1):1–14. doi: 10.1111/j.1471-4159.1982.tb10847.x. [DOI] [PubMed] [Google Scholar]

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