The sea lamprey recovers from spinal cord transection by a process that involves directionally specific regeneration of axons. The mechanisms underlying this speci ficity are not known, but they may involve molecular cues similar to those that guide the growth of spinal cord axons during development, such as netrins and semaphorins. To test the role of guidance cues in regeneration, we cloned netrin and its receptor UNC-5 from lamprey central nervous system (CNS) and studied their expression after spinal cord transection. In situ hybridization showed that (1) mRNA for netrin is ex pressed in the spinal cord, primarily in neurons of the lateral gray matter and in dor sal cells; (2) mRNA for UNC-5 is expressed in lamprey reticulospinal neurons; (3) fol lowing spinal cord transection, UNC-5 message was dramatically downregulated at two weeks, during the period of axon dieback; (4) UNC-5 message was upregulated at three weeks, when many axons are beginning to regenerate; and (5) axotomy-in duced expression of UNC-5 occurred primarily in neurons whose axons regenerate poorly. Because the UNC-5 receptor is thought to mediate the chemorepellent ef fects of netrins, netrin signaling may play a role in limiting or channeling the regen eration of certain neurons. These data strengthen the rationale for studying the role of developmental guidance molecules in CNS regeneration.
References
1.
David S., Aguayo AJAxonal elongation into peripheral nervous system "bridges" after central nervous system injury in adult rats. Science1981; 214:931-933.
2.
Caroni P., Schwab METwo membrane protein fractions from rat central myelin with inhibitory properties for neurite growth and fibroblast spreading. J Cell Biol1988; 106(4):1281-1288.
3.
Aguayo AJ, Rasminsky M., Bray GM, et al. Degenerative and regenerative responses of injured neurons in the central nervous system of adult mammals. Philos Trans R Soc Lond [Biol]1991; 331(1261):337-343.
4.
Richardson PM , Issa Vmk, Aguayo AJRegeneration of long spinal axons in the rat . J Neurocytol1984; 13:165-182.
5.
Vidal-Sanz M. , Bray GM, Villegas-Pérez MP, Thanos S, Aguayo AJ. Axonal regeneration and synapse formation in the superior colliculus by retinal ganglion cells in the adult rat. J Neurosci1987; 7:2894-2909.
6.
Schnell L., Schwab MEAxonal regeneration in the rat spinal cord produced by an antibody against myelin-associated neurite growth inhibitors. Nature1990; 343(6255):269-72.
7.
Bregman BS, Bernstein-Goral H.Both regenerating and late developing pathways contribute to transplant-induced anatomical plasticity after spinal cord transection. Exp Neurol1991; 112:49-63.
8.
Howland DR, Bregman BS, Tessler A, Goldberger ME. Transplants enhance locomotion in neonatal kittens whose spinal cords are transected: A behavioral and anatomical study. Exp Neurol1995; 135(2):123-145.
9.
Cheng H., Cao Y., Olson L.Spinal cord repair in adult paraplegic rats: Partial restoration of hind limb function. Science1996; 273: 510-513.
10.
Li Y., Field PM, Raisman G.Repair of adult rat corticospinal tract by transplants of olfactory ensheathing cells. Science1997; 277(5334):2000-2002.
11.
McKerracher L., David S., Jackson DL, Kottis V, Dunn RJ, Braun PE. Identification of myelin-associated glycoprotein as a major myetin-derived inhibitor of neurite growth. Neuron1994; 13(4):805-811.
12.
Spillmann AA , Bandtlow CE, Lottspeich F, Keller F, Schwab ME. Identification and characterization of a bovine neurite growth inhibitor (bN1-220). J Biol Chem1998; 273(30):19283-19293.
13.
Ghosh A., David S.Neurite growth-inhibitory activity in the adult rat cerebral cortical gray matter. J Neurobiol1997; 32(7):671-683.
14.
Snow DM, Lemmon V., Carrino DA, Caplan AI, Silver J. Sulfated proteoglyçans in astroglial barriers inhibit neurite outgrowth in vitro. Exp Neurol1990; 109(1):111-130.
15.
Fitch MT, Silver J.Glial cell extracellular matrix: boundaries for axon growth in development and regeneration. Cell Tissue Res1997; 290(2):379-384.
16.
Chen DF, Jhaveri S., Schneider GEIntrinsic changes in developing retinal neurons result in regenerative failure of their axons. Proc Natl Acad Sci USA1995 ; 92(16):7287-7291.
Becker T., Bernhardt RR, Reinhard E., Wullimann MF, Tongiorgi E, Schachner M. Readiness of zebrafish brain neurons to regenerate a spinal axon correlates with differential expression of specific cell recognition molecules. J Neurosci1998; 18(15): 5789-5803.
19.
Schnell L., Schneider R., Kolbeck R, Barde YA, Schwab ME. Neurotrophin-3 enhances sprouting of carticospinal tract during development and after adult spinal cord lesion [see comments]. Nature1994; 367(6459):170-173.
20.
Sawai H., Clarke DB, Kittlerova P, Bray GM, Aguayo AJ. Brain-derived neurotrophic factor and neurotrophin-4/5 stimulate growth of axonal branches from regenerating retinal ganglion cells. J Neurosci1996; 16(12):3887-3894.
21.
Ye JH, Houle JDTreatment of the chronically injured spinal cord with neurotrophic factors can promote axonal regeneration from supraspinal neurons. Exp Neurol1997; 143(1):70-81.
22.
Kobayashi NR , Fan DP, Giehl KM, et al. BDNF and NT-4/5 prevent atrophy of rat rubrospinal neurons after cervical axotomy, stimulate GAP-43 and Talphal-tubulin mRNA expression, and promote axonal regeneration. J Neurosci1997; 17(24):9583-95.
23.
Grill R., Murai K., Blesch A, Gage FH, Tuszynski MH. Cellular delivery of neurorrophin-3 promotes corticospinal axonal growth and partial functional recovery after spinal cord injury. J Neurosci1997; 17(14):5560-72.
24.
Liu Y., Kim D., Himes BT, et al. Transplants of fibroblasts genetically modified to express BDNF promote regeneration of adult rat rubrospinal axons and recovery of forelimb function. J Neurosci1999; 19(11):4370-4387.
25.
Cohen AH, Mackler SA, Selzer MEBehavioral recovery following spinal transection: Functional regeneration in the lamprey CNS. Acta Physiol Scand1988 ; 11(5):227-231.
26.
Rovainen CMRegeneration of Müller and Mauchner axons after spinal transection in larval lampreys. J Comp Neurol1976; 168(4):545-554.
27.
Selzer MEMechanisms of functional recovery and regeneration after spinal cord transection in larval sea lamprey. J Physiol (Lond)1978; 277(395):395-408.
28.
Wood MR, Cohen MJSynaptic regeneration in identified neurons of the lamprey spinal cords. Science1979; 206(4416): 344-347.
29.
Davis Grj, McClellan ADTime course of anatomical regeneration of descending brainstem neurons and behavioral recovery in spinal-transected lamprey. Brain Res1993; 602(1):131-137.
Davis GR, McClelland ADLong distance axonal regeneration of identified lamprey reticulospinal neurons . Exp Neurol1994; 127( 1):94-105.
32.
Mackler SA, Selzer MESpecificity of synaptic regeneration in the spinal cord of the larval sea lamprey. J Physiol (Lond)1987; 388(183):183-198.
33.
Yin HS, Mackler SA, Selzer MEDirectional specificity in the regeneration of lamprey spinal axons. Science1984; 224(4651): 894-896.
34.
Mackler SA, Yin HS, Selzer MEDeterminants of directional specificity in the regeneration of lamprey spinal axons. J Neurosci1986; 6(6):1814-1821.
35.
Tessier-Lavigne M., Goodman CSThe molecular biology of axon guidance. Science1996; 274(5290):1123-1133.
36.
Goodman CSMechanisms and molecules that control growth cone guidance. Ann Rev Neurosci1996; 19:341-77.
37.
Kennedy TE, Serafini T, de la Torre JR, Tessier-Lavigne M. Netrins are diffusible chemotropic factors for commissural axons in the embryonic spinal cord. Cell1994; 78(3):425-435.
38.
Serafini T. , Kennedy TE, Galko M] , et al. The netrins define a fomily of axon outgrowth-promoting proteins homologous to C. elegans UNC-6. Cell1994; 78(3):409-424.
39.
Keino-Masu K., Masu M., Hinck L., et al. Deleted in Colorectal Cancer (DCC) encodes a netrin receptor. Cell1996; 87(2):175-185.
40.
Hamelin M., Zhou Y., Su MW, Scott IM, Culotti JG. Expression of the UNC-5 guidance receptor in the touch neurons of C. elegans steers their axons dorsally. Nature1993; 364(6435):327-330.
41.
Leonardo ED , Hinck L., Masu M., et al. Vertebrate homologues of C. elegans UNC-5 are candidate netrin receptors. Nature1997; 386(6627):833-838.
42.
Ackerman SL , Kozak LP, Przyborski SA, et al. The mouse rostral cerebellar malformation gene encodes an UNC-5-like protein. Nature1997; 386(6627):838-42.
43.
Swedlow PE, Cooper EC, Lowenstein DHExpression of Deleted in colon cancer (DCC) in the adult rat brain. Society for Neuroscience Abstracts1997;23:325.
44.
Colicos MA, Baranes D., Aznar S., et al. Netrin expression in the adult mammalian brain. Society for Neuroscience Abstracts1997; 23:326.
45.
Volenec AV, Bhogal RK, Moorman JM, A. LR, T.P. F. Distribution of DCC mRNA in adult rat brain. Society for Neuroscience Abstracts1997; 23:325.
46.
Henikoff S., Henikoff JG, Alford W], Pietrokovski S.Automated construction and graphical presentation of protein blocks from unaligned sequences. Gene - COMBIS1995; 163:17-26.
47.
Swain GP, Jacobs AJ, Frei E, Selzer ME. A method for in situ hybridization in wholemounted lamprey brain: Neurofilament expression in larvae and adults . Exp Neurol1994; 126:256-269.
48.
Swain GPCNS Regeneration and Behavioral Recovery Following Spinal Cord Transection in the Sea Lamprey Petromyzon Marinus [PhD Dissertation]Northeastern University, 1989.
49.
Cohen AH, Baker MT, Dobrov TAEvidence for functional regeneration in the adult lamprey spinal cord following transection. Brain Res1989; 496(1-2):368-72.
50.
Lurie DI, Selzer MEAxonal regeneration in the adult lamprey spinal cord. J Comp Neurol1991b; 306(3):409-416.
51.
Roederer E. , Goldberg NH, Cohen MJModification of retrograde degeneration in transected spinal axons of the lamprey by applied DC current. J Neurosci1983; 3(1):153-160.
52.
Leung-Hagesteijn C., Spence AM, Stem BD, et al. UNC-5, a transmembrane protein with immunoglobulin and thrombospondin type 1 domains, guides cell and pioneer axon migrations in C. elegans. Cell1992; 71(2):289-99.
53.
Colavita A. , Culotti JGSuppressors of ectopic UNC-5 growth cone steering identify eight genes involved in axon guidance in Caenorhabditis elegans. Developmental Biology (Orlando)1998; 194(1):72-85.
54.
Hedgecock EM , Culotti JG, Hall DHThe unc-5, unc-6, and unc-40 genes guide circumferential migrations of pioneer axons and mesodermal cells on the epidermis in C. elegans . Neuron1990; 4(1):61-85.
55.
Pijak DS, Hall GF, Tenicki PJ, Boulos AS, Lurie DI, Selzer ME. Neurofilament spacing, phosphorylation, and axon diameter in regenerating and uninjured lamprey axons. J Comp Neurol1996; 368(4):569-581.
56.
Tetzlaff W. , Alexander SW, Miller FD, Bisby MA. Response of facial and rubrospinal neurons to axotomy: changes in mRNA expression for cytoskeletal proteins and GAP-43. J Neurosci1991; 11(8):2528-2544.
57.
Doster SK, Lozano AM, Aguayo AJ, Willard MB. Expression of the growth-associated protein GAP-43 in adult rat retinal ganglion cells following axon injury. Neuron1991; 6(4):635-47.
58.
Chong MS, Woolf CJ, Turmaine M, Emson PC, Anderson PN. Intrinsic versus extrinsic factors in determining the regeneration of the central processes of rat dorsal root ganglion neurons: the influence of a peripheral nerve graft. J Comp Neurol1996; 370(1): 97-104.
59.
Bisby MA, Tetzlaff W.Changes in cytoskeletal protein synthesis following axon injury and during axon regeneration. MolecularNeurobiology1992; 6(2-3):107-23.
Chong MS, Woolf CJ, Andrews P., et al. The downregulation of GAP-43 is not responsible for the failure of regeneration in freeze-killed nerve grafts in the rat. Exp Neurol1994; 129(2):311-320.
62.
Meyerhardt JA, Caca K., Eckstrand BC, et al. Netrin-1: interaction with deleted in colorectal cancer (DCC) and alterations in brain tumors and neuroblastomas . Cell Growth Differ1999; 10(1):35-42.
63.
Sterba G., Naumann W.Neural and glial secretion in the spinal cord of the lamprey (Lampetra planeri ). Acta Zool Stockholm1970; 51:159-168.
64.
Lauderdale JD, Davis NM, Kuwada JYAxon tracts correlate with netrin-la expression in the zebrafish embryo. Molecular & Cellular Neurosciences1997; 9(4):293-313.
65.
Goodhill GJMathematical guidance for axons. Trends Neurosci1998; 21(6):226-231.
66.
Luo YL, Raible D., Raper JACollapsin: a protein in brain that induces the collapse and paralysis of neuronal growth cones. Cell1993; 75(2):217-227.
67.
Luo Y., Shepherd I., Li J., Renzi MJ, Chang S, Raper JA. A family of molecules related to collapsin in the embryonic chick nervous system. Neuron1995; 14(6):1131-1140.
