Nerve growth factor (NGF) influences differentiation and proliferation of myogenic cells in vitro via TrKA

International Journal of Developmental Neuroscience - Tập 18 - Trang 869-885 - 2000
Mario Rende1, Emanuela Brizi1, Jim Conner2, Susan Treves3, Kathrin Censier4, Carlo Provenzano5, Giulio Taglialatela6, Pietro Paolo Sanna7, Rosario Donato1
1Department of Experimental Medicine and Biochemical Sciences, Section of Anatomy, University of Perugia School of Medicine, Via del Giochetto, 06125 Perugia, Italy
2Department of Neuroscience, University of California, San Diego, CA USA
3Department of Experimental Medicine and Diagnostic, Section of General Pathology, University of Ferrara, Ferrara, Italy
4Depertment of Anesthesia and Research, University of Basel, Basel, Switzerland
5Institute of General Pathology, Catholic University School of Medicine, Rome, Italy
6Department of Anatomy and Neuroscience, UTMB at Galveston, Galveston, TX, USA
7Department of Neuropharmacology, The Scripps Research Institute, La Jolla, CA, USA

Tóm tắt

AbstractClassic studies have established that muscle cells exert trophic actions on neurons of the developing peripheral nervous system through the production of neurotrophins. For this reason neurotrophins are also known as ‘target‐derived factors’. During differentiation, muscle cells also express some neurotrophin receptors, such as the low‐affinity p75 neurotrophin receptor, which binds all neurotrophins, and the high affinity tyrosine kinase receptor TrKA, nerve growth factor (NGF) transducing receptor. The functional roles of these receptors in muscle cells are still unclear and only fragmentary and controversial data are available regarding the responsiveness of muscle cells to NGF. The aim of the present study is to investigate the effects of NGF on cells of myogenic lineage. The rat myogenic cell line L6, primary cultures of adult human myoblasts, and the human rhabdomyosarcoma cell line TE‐671 were used in this study. As expected, all the three cell types expressed NGF, p75 and TrKA. NGF was expressed by L6 and primary myoblasts following differentiation, but it was constitutively expressed at high levels in the TE‐671 rhabdomyosarcoma cells. In L6 myoblasts, p75 receptor was expressed in myoblasts but not in myotubes early after plating; while some primary human myoblasts expressed it at all the time‐points tested. Some fusiform cells of the TE‐671 rhabdomyosarcoma cell line also expressed p75. TrKA was constitutively immunodetected in all the three cell lines, suggesting that these cells may respond to NGF. Addition of exogenous NGF increased the fusion rate of both primary and L6 myoblasts; as well as the proliferation of the slowly dividing primary myoblasts. Consistently, blocking the action of endogenously produced NGF with a specific neutralizing antibody decreased the percentage of fusion in both primary and L6 myoblasts. On the contrary, blocking the binding of NGF to p75 did not affect the percentage of fusion. Furthermore, neither exogenous NGF nor NGF‐ or p75‐neutralizing antibodies appeared to affect the rhabdomyosarcoma cells, which have a high proliferation rate and do not fuse. Pharmacological inhibition of TrKA signal transduction with K252a (in the nM range) and tyrphostin AG879 (in the low μM range) resulted in a dramatic dose‐dependent decrease in proliferation of all of the myogenic cell lines tested. Interestingly, this was especially evident in the rapidly dividing rhabdomyosarcoma cell line. The TrKA inhibitors also blocked fusion of L6 and primary myoblasts and induced morphological changes characterized by the flattening of the cells and a ‘spider‐like’ rearrangement of the intermediate filaments in all three cell lines with some minor differences. A transfection study showed that p75‐overexpressing L6 cells do not fuse and present changes in their morphology similar to the TrKA‐inhibitors treated L6 cells. These data support the notion that NGF expression in skeletal muscle is not only associated with a classical target‐derived neurotrophic function for peripheral nervous system neurons, but also with an autocrine action which affects the proliferation, fusion into myotubes, and cell morphology of developing myoblasts. The present data also suggest that these effects of NGF are mediated by TrKA receptors and that a sustained presence of NGF is needed for increase fusion into myotubes. Lastly, the dramatic anti‐proliferative effect of TrKA inhibitors on myogenic cells, and especially on the TE‐671 rhabdomyosarcoma cell line, suggests that pharmacological interference with NGF signal transduction could be effective in the control of these malignancies.

Tài liệu tham khảo

10.1098/rstb.1996.0037 10.1016/0304-3940(91)90418-S 10.1074/jbc.272.9.5936 10.1002/mus.880170304 10.1016/S0021-9258(18)48447-5 10.1016/0006-8993(87)91632-5 10.1002/jcp.1041440317 10.1002/(SICI)1097-4547(20000215)59:4<553::AID-JNR11>3.0.CO;2-4 10.1172/JCI993 10.1091/mbc.5.5.549 10.1016/0165-0270(95)00151-4 Conover J.C., 1997, Neurotrophin regulation of the developing nervous system analyses of knockout mice, Rev. Neurosci., 1, 13 10.1002/ana.410420604 10.1016/S0960-9822(95)00144-8 Del Valle M.E., 1992, Nerve growth factor receptor (NGFr) immunoreactivity in skeletal muscles of the rat, Cell. Mol. Biol., 38, 443 Delaporte C., 1987, Differentiation of human muscle cells in the presence of neuronal conditioned medium (NCM), nerve growth factor (NGF) and spinal cord cells (SC), Adv. Exp. Med. Biol., 209, 15 Donovan M.J., 1993, Immunohistochemical localization of Trk receptor protein in pediatric small round blue cell tumors, Am. J. Pathol., 143, 1560 10.1002/(SICI)1097-4652(199807)176:1<22::AID-JCP3>3.0.CO;2-A 10.1126/science.7770776 10.1083/jcb.123.2.455 10.1021/jm00130a020 10.1002/jnr.490420104 10.1038/363266a0 10.1006/dbio.1995.1019 10.1016/0169-328X(89)90007-7 10.1177/096368979800700609 10.1523/JNEUROSCI.08-02-00715.1988 10.1016/S0959-8049(97)00210-4 Lee K.F., 1992, Targeted mutation of the gene encoding the low affinity NGF receptor p75 leads to deficits in the peripheral sensory nervous system, Cell, 5, 737, 10.1016/0092-8674(92)90286-L 10.1002/jez.1401160206 10.1002/jez.1401230203 10.1046/j.1471-4159.1995.64041780.x 10.1038/319743a0 10.1046/j.1471-4159.1996.66010105.x 10.1073/pnas.90.17.7961 10.1210/me.10.3.272 Nelson R.J., 1998, Behaviour in mice with targeted disruption of single genes, Neurosci. Biobehav. Rev., 3, 453, 10.1016/S0149-7634(97)00053-5 10.1097/00019606-199702000-00003 10.1021/bi00068a024 OuJ.‐Y. ChambersJ. WheelerE.F. Myoblasts differentiation is accompanied by expression of the NGF receptors: effects of NGF on the proliferation and differentiation of myoblasts. In:Proceedings of the 29th Annual Meeting of the Society for Neuroscience 1999 Abs. 94.15. Pich E.M., 1998, Transgenic mice in drug dependence research, Ann. Med., 4, 390, 10.3109/07853899809029939 Radeva V., 1978, Effects of the nerve growth factor (NGF) on certain non‐neuronal elements, Agressologie, 19, 99 RendeM. BriziE. BrunoR. ProvenzanoC. ConnerJ. Nerve growth factor (NGF) stimulates myoblast fusion into myotubes in a rat myogenic cell line (L6): a novel autocrine role for NGF in muscle development. In:Proceedings of the 28th Annual Meeting of Society for Neuroscience 1998 Abs. 801.10. 10.1023/A:1003851024732 10.1016/S0006-8993(98)00396-5 10.1126/science.277.5329.1097 10.1038/sj.bjp.0701345 10.1016/0896-6273(92)90183-E 10.1016/0014-4827(86)90574-4 10.1002/(SICI)1097-4652(199807)176:1<10::AID-JCP2>3.0.CO;2-B 10.1002/(SICI)1097-4598(199803)21:3<404::AID-MUS17>3.0.CO;2-Z 10.1002/(SICI)1097-0142(19981015)83:8<1626::AID-CNCR19>3.0.CO;2-Y Tapley P., 1992, K252a is a selective inhibitor of the tyrosine protein kinase activity of the trk family of oncogenes and neurotrophin receptors, Oncogene, 7, 371 10.1111/j.1460-9568.1993.tb00526.x 10.1523/JNEUROSCI.15-07-04796.1995 10.1007/BF02303537 10.1523/JNEUROSCI.12-03-00930.1992 10.1002/(SICI)1096-9861(19980222)391:4<407::AID-CNE1>3.0.CO;2-4
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International Journal of Developmental Neuroscience

Tập 18

869-885

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