Denervation of CNS neurons and peripheral organs is a consequence of traumatic SCI. Intraspinal transplantation of embryonic CNS neurons is a potential strategy for reinnervating these targets. Neural progenitor cell lines are being investigated as alternates to embryonic CNS neurons. RN33B is an immortalized neural progenitor cell line derived from embryonic rat raphé nuclei following infection with a retrovirus encoding the temperature-sensitive mutant of SV40 large T-antigen. Transplantation studies have shown that local epigenetic signals in intact or partially neuron-depleted adult rat hippocampal formation or striatum direct RN33B cell differentiation to complex multipolar morphologies resembling endogenous neurons. After transplantation into neuron-depleted regions of the hippocampal formation or striatum, RN33B cells were relatively undifferentiated or differentiated with bipolar morphologies. The present study examines RN33B cell differentiation after transplantation into normal spinal cord and under different lesion conditions. Adult rats underwent either unilateral lesion of lumbar spinal neurons by intraspinal injection of kainic acid or complete transection at the T10 spinal segment. Neonatal rats underwent either unilateral lesion of lumbar motoneurons by sciatic nerve crush or complete transection at the T10 segment. At 2 or 6-7 wk postinjury, lacZ-labeled RN33B cells were transplanted into the lumbar enlargement of injured and age-matched normal rats. At 2 wk posttransplantation, bipolar and some multipolar RN33B cells were found throughout normal rat gray matter. In contrast, only bipolar RN33B cells were seen in gray matter of kainic acid lesioned, sciatic nerve crush, or transection rats. These observations suggest that RN33B cell multipolar morphological differentiation in normal adult spinal cord is mediated by direct cell-cell interaction through surface molecules on endogenous neurons and may be suppressed by molecules released after SCI. They also indicate that the fate of immortalized neural progenitor cell lines in injured CNS must be stringently characterized.
AdamsF.S.; La RossaF.G.; KumarS.; Edwards-PrasadJ.; KentoriS.; VernadakisA.; FreedC.R.; PrasadK.N.Characterization and transplantation of two neuronal cell lines with dopaminergic properties.Neurochem. Res.21: 619–627; 1996.
2.
BakhitC.; ArmaniniM.; WongW.L.; BennettG.L.; WrathallJ.R.Increase in nerve growth factor-like immu-noreactivity and decrease in choline acetyltransferase following contusive spinal cord injury.Brain Res.554: 264–271; 1991.
3.
BarrettC.P.; GuthL.; DonatiE.J.; KrikorianJ.C.Astroglial reaction in the gray matter of lumbar segments after midthoracic transection of the adult rat spinal cord.Exp. Neurol.73: 365–377; 1981.
4.
BenvenisteE.N.Cytokine expression in the nervous system. In: KeaneR.W.; HickeyW.F., eds. Immunology of the Nervous System.New York: Oxford University Press;1997: 419–459.
5.
BjörklundA.; NornesH.; GageF.H.Cell suspension grafts of noradrenergic locus coeruleus neurons in rat hippocampus and spinal cord: Reinnervation and transmitter turnover.Neuroscience18: 685–698; 1986.
6.
BregmanB.S.Recovery of function after spinal cord injury: Transplantation strategies. In: DunnettS.B.; BjörklundA., eds. Functional Neural Transplantation.New York: Raven Press;1994: 489–529.
7.
BregmanB.S.; Kunkel-BagdenE.; ReierP.J.; DaiH.N.; McAteeM.; GaoD.Recovery of function after spinal cord injury: Mechanisms underlying transplant-mediated recovery of function differ after spinal cord injury in newborn and adult rats.Exp. Neurol.123: 3–16; 1993.
8.
BrundinP.; BjörklundA.Survival, growth and function of dopaminergic neurons grafted to the brain.Prog. Brain Res.71: 293–308; 1987.
BuchananJ.T.; NornesH.O.Transplants of embryonic brainstem containing the locus coeruleus into spinal cord enhance the hindlimb flexion reflex in adult rats.Brain Res.381: 225–236; 1986.
BungeR.P.; PuckettW.R.; BecerraJ.L.; MarcilloA.; QuencerR.M.Observations on the pathology of human spinal cord injury: A review and classification of 22 new cases with details from a case of chronic spinal cord compression with extensive focal demyelination. In: SeilF.J., ed. Neural regeneration.New York: Raven Press; 1992: Advances in Neurology 59: 75–89.
13.
CattaneoE.; MagrassiL.; ButtiG.; SantiL.; GiavazziA.; PezzottaS.A short term analysis of the behavior of conditionally immortalized neuronal progenitors and primary neuroepithelial cells implanted into the fetal rat brain.Dev. Brain Res.83: 197–208; 1994.
14.
DasG.D.Neural transplantation in the spinal cord of adult rats.Conditions, survival, cytology and connectivity of the transplants. J. Neurol. Cytol.62: 191–210; 1983.
15.
EatonM.J.; WhittemoreS.R.Autocrine secretion of BDNF enhances survival and serotonergic differentiation of raphe precursor cells transplanted into the adult rat CNS.Exp. Neurol.140: 105–114; 1996.
16.
FarooqueM.; BadonicT.; OlssonY.; HoltzA.Astrocytic reaction after graded spinal cord compression in rats: Immunohistochemical studies on glial fibrillary acidic protein and vimentin.J. Neurotrauma12: 41–52; 1995.
FosterG.A.; BrodinE.; GageF.H.; MaxwellD.J.; RobertsM.H.T.; SharpT.Restoration of function to the denervated spinal cord after implantation of embryonic 5-HT- and substance P-containing raphe neurons. In: DunnettS.B.; RichardsS.-J., eds. Neural transplantation: From molecular basis to clinical applications. Progress in Brain Research, vol. 82. New York: Elsevier;1990: 247–259.
19.
FrisenJ.; HægerstarandA.; FriedK.; CullheimS.; RislingM.Adhesive/repulsive properties in the injured spinal cord: Relation to myelin phagocytosis by invading macrophages.Exp. Neurol.129: 183–193; 1994.
20.
FrisenJ.; VergeV.K.; CullheimS.; PerssonH.; FriedK.; MiddlemasD.S.; HunterT.; HökfeltT.; RislingM.Increased levels of trkB mRNA and trkB protein-like immunoreactivity in the injured rat and cat spinal cord.Proc. Natl. Acad. Sci. USA89: 11282–11286; 1992.
21.
FrisénJ.; JohanssonC.B.; TörökC.; RislingM.; LendahlU.Rapid, widespread, and longlasting induction of nestin contributes to the generation of glial scar tissue after CNS injury.J. Cell Biol.131: 453–464; 1995.
22.
GageF.H.; RayJ.; FisherL.J.Isolation, characterization, and use of stem cells from the CNS.Annu. Rev. Neurosci.18: 159–192; 1995.
23.
GiordanoM.; TakashimaH.; PoltorakM.; GellerH.M.; FreedW.J.Constitutive expression of glutamic acid decarboxylase (GAD) by striatal cell lines immortalized using the tsA58 allele of the SV40 large T antigen.Cell Transplant.5: 563–575; 1996.
24.
HofferB.J.; OlsonL.Ethical issues in brain-cell transplantation.Trends Neurosci.14: 384–388; 1991.
25.
HorvatJ.-C.; Baillet-DerbinC.; YeJ.H.; RhrichF.; AffaneF.Co-transplantation of embryonic neural tissue and autologous peripheral nerve segments to severe spinal cord injury of the adult rat. Guided axogenesis from transplanted neurons.Restor. Neurol. Neurosci.2: 289–298; 1991.
26.
KoshinagaM.; SanonH.R.; WhittemoreS.R.Altered acidic and basic fibroblast growth factor expression following spinal cord injury.Exp. Neurol.120: 32–48; 1993.
27.
KoshinagaM.; WhittemoreS.R.The temporal and spatial activation of microglia in fiber tracts undergoing anterograde and retrograde degeneration following spinal cord injury.J. Neurotrauma12: 209–222; 1995.
28.
LundbergC.; WinklerC.; WhittemoreS.R.; BjörklundA.Conditionally immortalized neural progenitor cells grafted to the striatum exhibit site-specific neuronal differentiation and establish connections with the host globus pallidus.Neurobiol. Dis.3: 33–50; 1996.
29.
McKeonR.J.; SchreiberR.C.; RudgeJ.S.; SilverJ.Reduction of neurite outgrowth in a model of glial scarring following CNS injury is correlated with the expression of inhibitory molecules on reactive astrocytes.J. Neurosci.11: 3398–3411; 1991.
MoormanS.J.; WhalenL.R.; NornesH.O.A neurotransmitter specific functional recovery mediated by fetal implants in the lesioned spinal cord of the rat.Brain Res.508: 247–259; 1990.
32.
NolanG.P.; FieringS.; NicholasJ.F.; HerzenbergL.A.Fluorescence-activated cell analysis and sorting of viable mammalian cells based on a B-D-galactosidase activity after transduction of Escherichia coli lacZ. Proc.Natl. Acad. Sci. USA85: 2603–2607; 1988.
33.
NothiasF.; PeschanskiM.Homotypic fetal transplants into an experimental model of spinal cord neurodegeneration.J. Comp. Neurol.301: 520–534; 1990.
34.
NygrenL.-G.; OlsonL.; SeigerÅ.Monoaminergic reinnervation of the transected spinal cord by homologous fetal brain grafts.Brain Res.129: 227–235; 1977.
35.
OniferS.M.; WhiteL.A.; WhittemoreS.R.; HoletsV.R.In vitro strategies for identifying transplanted primary CNS tissue and neuronal cell lines.Cell Transplant.2: 131–149; 1993.
36.
OniferS.M.; WhittemoreS.R.; HoletsV.R.Variable morphological differentiation of a raphe-derived neuronal cell line following transplantation into the adult rat CNS.Exp. Neurol.122: 130–142; 1993.
37.
PrivatA.; MansourH.; GeffardM.Transplantation of fetal serotonin neurons into the transected spinal cord of adult rats: Morphological development and functional influence. In: GashD.M.; SladekJ.R.Jr., eds. Transplantation into the mammalian CNS. Progress in Brain Research, vol. 78. New York: Elsevier;1990: 155–166.
38.
RajaofetraN.; PoulatP.; MarlierL.; SandillonF.; DrianM.J.; KönigN.; FamoseF.; VerschuereB.; GouyD.; GeffardM.; PrivatA.Transplantation of embryonic se-rotonin immunoreactive neurons into the transected spinal cord of adult monkey (Macaca fascicularis).Brain Res.572: 329–334; 1992.
39.
ReierP.J.; StokesB.T.; ThompsonF.J.; AndersonD.K.Fetal cell grafts into resection and contusion/compression injuries of the rat and cat spinal cord.Exp. Neurol.115: 177–188; 1992.
40.
ReynoldsM.; BrunelloN.; MocchettiI.; WrathallJ.R.Localization of nerve growth factor mRNA in contused rat spinal cord by in situ hybridization.Brain Res.559: 149–153; 1991.
41.
RislingM.; FriedK.; LindáH.; CullheimS.; MeierM.Changes in nerve growth factor receptor-like immunoreactivity in the spinal cord after vertral funiculus lesion in adult cats.J. Neurocytol.21: 79–93; 1992.
42.
RomanesG.J.Motor localization and the effects of nerve injury on the ventral horn cells of the spinal cord.J. Anat.80: 117–131; 1946.
43.
RoudetC.; Gimenez y RibottaM.; PrivatA.; FeuersteinC.; SavastaM.Intraspinal noradrenergic-rich implants reverse the increase of a ladenoceptors densities caused by complete spinal cord transection or selective chemical denervation: A quantitative autoradiographic study.Brain Res.677: 1–12; 1995.
44.
SchwabM.E.; KapfhammerJ.P.; BandtlowC.E.Inhibitors of neurite growth.Annu. Rev. Neurosci.16: 565–595; 1993.
45.
SeigerÅ.Preparation of immature central nervous system regions for transplantation. In: BjörklundA.; SteneviU., eds. Neural Grafting in the Mammalian CNS.Amsterdam: Elsevier;1985: 71–77.
46.
ShihabuddinL.S.; BrunschwigJ.-P.; HoletsV.R.; BungeM.B.; WhittemoreS.R.Induction of mature neuronal properties in immortalized neuronal precursor cells following grafting into the neonatal CNS.J. Neurocytol.25: 101–111; 1996.
47.
ShihabuddinL.S.; HertzJ.A.; HoletsV.R.; WhittemoreS.R.The adult CNS retains the potential to direct region-specific differentiation of a transplanted neuronal precursor cell line.J. Neurosci.15: 6666–6678; 1995.
48.
ShihabuddinL.S.; HoletsV.R.; WhittemoreS.R.Selective hippocampal lesions differentially affect the phenotypic fate of transplanted neuronal precursor cells.Exp. Neurol.139: 61–72; 1996.
49.
ShimohamaS.; RosenbergM.B.; FaganA.M.; WolffJ.A.; ShortM.P.; BreakefieldX.O.; FriedmannT.; GageF.H.Grafting genetically modified cells into the rat brain: Characteristics of E. coli β-galactosidase as a reporter gene.Mol. Brain Res.5: 271–278; 1989.
50.
SieradzanK.; VrbováG.Replacement of missing motoneurons by embryonic grafts in the rat spinal cord.Neuroscience31: 115–130; 1989.
51.
SteinbuschH.W.M.; NiewenhuysR.The raphe nuclei of the rat brainstem: A cytoarchitectonic and immunohisto-chemical study. In: EmsonP.C., ed. Chemical Neuroanatomy.New York: Raven Press;1983: 131–207.
52.
SugarO.; GerardR.W.Spinal cord regeneration in the rat.J. Neurophysiol.3: 1–19; 1940.
53.
TagoH.; McGeerP.L.; McGeerE.G.; AkiyamaH.; HershL.B.Distribution of choline acetyltransferase immunopositive structures in the rat brainstem.Brain Res.495: 271–297; 1989.
54.
TesslerA.; HimesB.T.; HouleJ.; ReierP.J.Regeneration of adult dorsal root axons into transplants of embryonic spinal cord.J. Comp. Neurol.270: 537–548; 1988.
55.
WhiteL.A.; KeaneR.W.; WhittemoreS.R.Differentiation of an immortalized CNS neuronal cell line decreases their susceptibility to cytotoxic T lymphocyte cell lysis in vitro.J. Neuroimmunol.49: 135–143; 1994.
56.
WhiteL.A.; ReebenM.; SaarmaM.; WhittemoreS.R.Transcriptional regulation of neurofilament expression by protein kinase A.J. Neurosci. Res.47: 242–252, 1997.
57.
WhiteL.A.; WhittemoreS.R.Immortalization of raphe neurons: An approach to neuronal function in vitro and in vivo.J. Chem. Neuroanat.5: 327–330; 1992.
58.
WhittemoreS.R.; SnyderE.Y.The physiological relevance and functional potential of central nervous system-derived cell lines.Mol. Neurobiol.12: 13–38; 1996.
59.
WhittemoreS.R.; WhiteL.A.Target regulation of neuronal differentiation in a temperature-sensitive cell line derived from medullary raphe.Brain Res.615: 27–40; 1993.
60.
WictorinK.; BjörklundA.Axon outgrowth from grafts of human embryonic spinal cord in the lesioned adult rat spinal cord.Neuroreport3: 1045–1048; 1992.
61.
WidnerH.; BrundinP.Immunological aspects of grafting in the mammalian central nervous system. A review and speculative synthesis.Brain Res. Rev.13: 287–324; 1988.
62.
YakovleffA.; CabelguenJ.-M.; OrsalD.; Gimenez y RibottaM.; RajaofetraN.; DrianM.-J.; BusselB.; PrivatA.Fictive motor activities in adult chronic spinal rats transplanted with embryonic brainstem neurons.Exp. Brain Res.106: 69–78; 1995.
