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Cell Tissue Res (1998) 294:19 Springer-Verlag 1998

REVIEW
Christine C. Stichel Hans Werner Mller
The CNS lesion scar: new vistas on an old regeneration barrier
Received: 19 June 1998 / Accepted: 23 July 1998
Studies in the authors' laboratory were supported by the Deutsche
Forschungsgemeinschaft (SFB 194/B5 and Graduiertenkolleg) and
a Heisenberg Stipendium to C.C.S.
C.C. Stichel (
)
) H.W. Mller
Molecular Neurobiology Laboratory, Department of Neurology,
Heinrich-Heine University, Moorenstrasse 5,
D-40225 Dsseldorf, Germany
Tel.: +49 0211 81 17822; Fax: +49 0211 81 18485;
e-mail: stichel@uni-duesseldorf.de
Abstract Scar formation represents a reaction of nervous
tissue to any form of physical injury. Research over the
past decade has demonstrated that the scar composed of
glial cells and several extracellular matrix molecules con-
stitutes an obstacle to axon regeneration in the CNS. This
review briefly summarizes the current knowledge on (a)
the structural and functional features of the lesion scar
and (b) the development of therapeutic interventions to
override this regeneration barrier.
Key words Axonal regeneration CNS injury
Growth inhibition Gliosis Extracellular matrix
Basal membrane Therapeutic approaches
Introduction
Mechanical injury to the adult mammalian CNS always
results in the formation of a scar. This cardinal histopath-
ological reaction at the site of lesion was first noticed by
Ramn y Cajal (1928), who also postulated a causal role
of this response in axonal regeneration failure: Neither is
it rare to see conductors that turn back on encountering
the scar, near which they accumulate, forming complicat-
ed plexuses, as though the edge of the nerve presented an
obstacle that the cones find it very difficult to cross.
Glial cell transplantation studies and the recent concept
of an extrinsic determination of axonal regeneration have
renewed the interest in lesion scar influences in axonal re-
growth. Over the past decade considerable advances have
been made in identifying the scar components and in de-
fining those factors that are responsible for axonal growth
arrest. Most importantly, recent studies have led to signif-
icant progress in the development of new concepts to
overcome the regeneration-inhibiting scar. They demon-
strate that the reestablishment of injured fiber tracts
across the lesion site and through the original pathway
is a goal that can be achieved. These new regeneration-
stimulating strategies hold promise as therapeutic ap-
proaches for traumatic injuries and neurodegenerative dis-
eases of brain and spinal cord.
Cellular and molecular composition of lesion scars
Injury to the CNS leads, like in all other organs of the
body, to the formation of a scar located at the site of le-
sion. This rapid and complex process includes both meso-
dermal cells, such as fibroblasts, endothelial and hema-
togenous cells, and glial cells, such as astrocytes and mi-
croglia/macrophages (Table 1). After lesion many glial
cells start to proliferate and change their morphological
and functional properties. Together with exudated serum
factors they induce the second major type of histopatho-
logical alteration within the scar area: a profound change
in the composition of the local extracellular matrix
(ECM) (Table 1).
Although different glial cells participate in CNS scar
formation, the reactive astrocytes play the predominant
role (Lindsay 1986). The vigorous response of astrocytes
Table 1 Lesion scar components
Cellular Extracellular
Reactive astrocytes Collagen IV
Microglia/macrophages Laminin
Endothelial cells Fibronectin
Fibroblasts Chondroitin sulfate proteoglycans
Heparan sulfate proteoglycans (perlecan)
Keratan sulfate proteoglycans
Tenascin C
Thrombospondin
Basal membrane
2
as reflected by morphological changes and by an increase
in the synthesis of glial fibrillary acidic protein (GFAP) is
a stereotyped reaction to any type of CNS insult (Fig. 1F).
It is also well conserved across a variety of species and
extends even in areas distant from the lesion interface
(Frank and Wolburg 1996; Maxwell et al. 1990; McLoon
1986; Puckett et al. 1997; Sivron et al. 1990; Stichel and
Mller 1994a). The reactive astrocytes form a dense web
of interdigitated processes, called (astro)gliosis, which
fills the space vacated by the dead or dying cells. These
structural changes are accompanied by diverse physiolog-
ical alterations. While the upregulation of GFAP is a
global response of astrocytes (Fig. 1F), various other mol-
ecules have been identified which are up- or downregulat-
ed in all or only in subsets of reactive astrocytes (Fig. 1J
L). Interestingly, many of these molecules are already ex-
pressed during normal development and are downregulat-
ed when development is complete (Brodkey et al. 1993;
Aubert et al. 1995). There are excellent reviews with a
comprehensive listing of astrocytic molecules already
available (Eddleston and Mucke 1993; Ridet et al.
1997) that need not be rereviewed in this paper. However,
one important group of astrocytic proteins should be men-
tioned in a paper dealing with axonal regeneration. These
are the glycoproteins laminin (Fig. 1G), fibronectin, ten-
ascin C (Fig. 1J) and the large family of proteoglycans
(Fig. 1K). In vitro studies, showing their various and
sometimes conflicting growth-modulating effects as well
as their neurotrophic activities, imply that they play an
important role in the regeneration process (for reviews
see Faissner 1997; Fawcett 1997; Hke and Silver 1996;
Mller et al. 1996).
During recent years evidence has accumulated that the
astrocytic response is heterogeneous and differs depend-
ing on the nature of the injury, the proximity to the lesion
and the microenvironment at the lesion site (David and
Ness 1993; Fernaud-Espinosa et al. 1993; Hatten et al.
1991). Accordingly, gliosis can be classified as anisomor-
phic or isomorphic, based on whether or not the eliciting
stimulus is an open injury or not. In the case of anisomor-
phic injury, astrocytes proliferate and express a different
set of molecules than their isotropic counterparts
(Fernaud-Espinosa et al. 1993; Ridet et al. 1997;
Bovolenta et al. 1992). Moreover, their functional proper-
ties may also depend on the distance to the lesion site.
Thus, reactive astrocytes in the immediate vicinity of a le-
sion generally display more excessive changes in gene ex-
pression patterns, e.g., increased expression of ECM mol-
ecules. This astrocytic heterogeneity might reflect differ-
ences in the factors or signals that induce the reactive
changes and depend on the extent of damage or degener-
ation.
A similar heterogeneous response has been reported
for the second prominent cell type of the scar, the micro-
glial cells. They are rapidly activated in a wide area
around the lesion site and undergo a profound change in
cell shape and phenotype (Fig. 1D) (Perry and Gordon
1988; Streit et al. 1988). Microglial cells and their phago-
cytic form, the macrophages (Fig. 1E), are found to se-
crete a number of cytokines, bioactive lipids, coagulation
factors, reactive oxygen intermediates and also neurotro-
phic factors (Auger and Ross 1992; Elkabes et al. 1996;
Chamak et al. 1994). The expression of these molecules
depends on the proximity to the lesion site, but is always
more prominent in the immediate surroundings of the le-
sion center. Within the latter these cells intermingle with
several blood vessels. Indeed, stimulated by various an-
giogenic factors (Klagsbrun and D'Amore 1991), endo-
thelial cells start to proliferate, to migrate and to synthe-
size matrix proteins (Landis 1994). After injury the mean
number of capillary profiles in the lesioned area is twice
that of uninjured regions and the new blood vessels exhib-
it an increased total luminal area and supernumerary lay-
ers of basal lamina (Jaeger and Blight 1997; Blight 1991;
Kalaria and Pax 1995).
All of these cells produce a variety of cytokines,
growth factors, cell adhesion and matrix molecules after
injury. Most of these molecules are soluble and are liber-
ated into the extracellular space (ECS) (Fig. 1GM)
(Nicholson and Sykova 1998). Formerly referred to as
ground substance, the matrix of this ECS, the so-called
extracellular matrix (ECM), represents a sort of reservoir,
in which the secreted factors organize into a superstruc-
ture of macromolecules (Rutka et al. 1988; Faissner
1997; Carbonetto 1984) and imbed the neurons and glial
cells. However, the ECM is a dynamic structure and
changes in composition, density and architecture after in-
jury. One of the most dominant histopathological ECM
features formed is the basal membrane (BM) (Timpl
and Dziadek 1986; Choi 1997; Yurchenco and O'Rear
1994; Paulsson 1992). Within the lesion site extensive de-
posits of BM are laid down (Fig. 1H) (Feringa et al. 1980;
Berry et al. 1983; Bernstein et al. 1985; Maxwell et al.
1984), the density of BM around blood vessels increases
(Kalaria and Pax 1995) and even astrocytic processes be-
come partially covered by thin BM sheets (Jaeger and
Blight 1997). The BM shows a three-layered structure
with the basal lamina as one prominent layer. The mole-
cular architecture of the BM is created by glycoprotein
and proteoglycan protomers, with two independent net-
works one formed by collagen IV (Coll IV) (Fig. 1H)
and the other by laminin (LN) (Fig. 1G). While it is
known from studies in the kidney and tumor tissue that
within this Coll IV/LN scaffold 50 or more constituents
may be anchored, the list of well-characterized BM com-
ponents, especially in the CNS, is rather short. Besides
Coll IV and LN, the major BM components, entactin/nid-
ogen, fibronectin, BM-40/osteonectin/SPARC (secreted
protein acidic and rich in cysteine), fibulin-1 and the hep-
aran sulfate proteoglycan perlecan were identified and
also the presence of chondroitin sulfate proteoglycans in
Fig. 1AM Summary of cellular and molecular changes in the
proximal postcommissural fornix and the transection site at 9
14 days post-lesion. At this time point the spontaneously sprouting
fibers reached the lesion site (A). Scale bars 50 m (reprinted with
permission from Stichel-Gunkel 1997)
3
4
brain BM has been shown (Couchman et al. 1984; Stichel
et al. 1998).
The proteins LN and Coll IV are major mediators of
contacts between BM and cells. Many but not necessarily
all signals provided by these two proteins are transduced
by the integrin class of cellular receptors (Hall et al. 1990;
Kramer and Marks 1989; Joosten 1996). Thus, it is known
that astrocytes attach to the BM and both together form
the so-called glia limitans, which surrounds the blood ves-
sels and covers the CNS.
Finally, the lesion scar might be characterized as a
gross cellular and molecular rearrangement of tissue mor-
phology with a series of common motifs (i.e., the overex-
pression of GFAP or the formation of a BM) to which
several other specific factors are added, depending on
the type of injury and the brain region damaged. Today,
we are far from having a complete description of all scar
components. However, more and more pieces are becom-
ing available and give rise to hope that this scar puzzle
will be completed soon.
Lesion scar functions
Having explored the dynamics and cellular and molecular
features of the lesion scar, the next issue to be addressed
is that of the functional consequences of such a tremen-
dous reorganization of tissue. What is the purpose of this
profound modulation, and does the CNS benefit from
scarring? The answer is, it certainly does. The lesion scar
reestablishes the integrity of the CNS by sealing it off
from the external environment. The latter effect allows re-
establishment of the diffusional barrier between the CNS
and its environment and prevents the CNS from infection
and further spread of damage. The BM exhibits contrac-
tile properties and therefore another job of scarring a
wound is to bring the wound margins toward one another
in order to shrink the lesion cavity and to reconstruct the
damaged parts. The lesion scar is then heavily invaded by
epithelial cells that reorganize the blood vessels of the
scar tissue. The resulting increased revascularization pro-
vides the nutritional, trophic and metabolic support essen-
tial for wound healing.
Scarring and regeneration failure
The above-mentioned beneficial effects of the scarring
process contrast with its clear inhibitory effects on the re-
generation of interrupted projections. In a long series of
experiments stretching back many years the lesion scar
has been identified as one critical element that impedes
axonal regeneration in the adult mammalian CNS (Ramn
Fig. 2A, B The influence of Schwann cell suspension implants on
axonal regeneration in the transected postcommissural fornix. A In
animals with transection only axons sprout up to the lesion site (ar-
row), where they stop. B In animals with an additional Schwann cell
implant the axons cross the lesion/implantation site (arrow) and en-
ter their former pathway, the distal fornix stump (d distal, p proxi-
mal). Scale bars 100 m
5
y Cajal 1928; Reier et al. 1983; Guth et al. 1983). After
lesion CNS axons start to sprout over short distances,
but almost all of them stop abruptly at the lesion scar bor-
der and fail to traverse it (Figs. 1A, 3A,B) (Stichel and
Mller 1994b; Blaugrund et al. 1993; Richardson et al.
1982; Schnell and Schwab 1993). Only single axons ei-
ther emit short sprouts into the scar, where they end local-
ly (Frisn et al. 1993; Li and Raisman 1995), or they are
deflected and bypass the lesion scar via bridges of con-
nective tissue, blood vessels or surviving tissue (Clemente
1955; Guth et al. 1983; Krger et al. 1986).
Reasons offered to explain the inhibitory nature of the
scar include (a) the accumulation of growth-inhibitory
molecules (molecular barrier) and (b) the formation of a
dense, impenetrable matrix within this area (physical bar-
rier).
Due to their dominant role in scar formation and their
diverse physiological changes after lesion, reactive astro-
cytes are suggested to play a key role in the formation of a
regeneration barrier. They (a) express several potential
growth-inhibitory molecules, such as tenascin C or
proteoglycans (Ajemian et al. 1994; Grierson et al.
1990; Bartsch et al. 1992; Stichel et al. 1995a; Canning
et al. 1996; Geisert et al. 1992), and (b) are able to form
a dense network with a strikingly high number of gap
junctions (Alonso and Privat 1993b). The fact that axons
succeed in entering and elongating within dense astrogli-
otic tissue (Alonso and Privat 1993b; Lips et al. 1995;
Stichel et al. 1994b; Alonso and Privat 1993a; Frisn et
al. 1993; Li and Raisman 1995) or a proteoglycan-rich
perilesional area (Lips et al. 1995; Stichel et al. 1995a)
encourages the view that reactive astrocytes and their
products are not a priori impermeable barriers for axons.
The BM, which is always deposited in the center of a
scar, might be another candidate responsible for abortive
axonal regeneration. Due to its dense architecture and in-
teraction with several growth-modulating molecules
(Timpl and Dziadek 1986), the BM might either create
a mechanical obstacle, a molecular barrier or both for re-
growing axons.
Recent experiments in our laboratory have renewed in-
terest in the lesion-induced BM formation and have cast
new light on its role in regeneration failure. In our lesion
model, the mechanically transected postcommissural for-
nix, sprouting fornix axons cross astrogliotic tissue rich in
Fig. 3 Sections reacted for neurofilament immunohistochemistry
show that in untreated animals (A, B) sprouting axons stop abruptly
at the lesion site (arrow), while in animals with dipyridyl injection
(C, D) axons succeed in crossing the lesion site (arrow) and enter
the distal fornix stump (d distal, p proximal). Scale bars 100 m
(A, C), 50 m (B, D)
6
both chondroitin/keratan sulfate proteoglycans and tenas-
cin C (Stichel et al. 1995; Stichel and Mller 1994b; Lips
et al. 1995), but abruptly stop growing at the BM in the
lesion center. The BM formed within the 1st week postle-
sion and penetrated the full depth of the lesion. It ap-
peared as a parallel array of BM sheets aligned perpendic-
ular to the course of the tract and intermingled with sev-
eral blood vessels. Confocal microscopy of double-la-
beled sections revealed that axons either arrest close to
BM deposits or make hook-like turns and stop some dis-
tance from the BM (Fig. 3A,B). The temporal coincidence
of BM maturation and the arrival of regrowing axons as
well as the close apposition of growth-arrested axons at
the BM sheets suggest that the BM is the inhibitory con-
stituent of the impermeable lesion scar.
Strategies to overcome the lesion scar
In the past a substantial number of different experimental
approaches have been made to drive regrowing axons,
e.g., by the use of neurotrophins (Bregman et al. 1997;
Houle et al. 1996; Tuszynski et al. 1996), the transplanta-
tion of nerve grafts (Vidal-Sanz et al. 1987; Cheng et al.
1996), Schwann cells (Harvey et al. 1995; Paino et al.
1994; Stichel et al. 1996; Xu et al. 1995), olfactory en-
sheathing cells (Ramon-Cuto et al. 1998; Ramon-Cuto
and Nieto-Sampedro 1994; Li et al. 1997) and micro-
glia/macrophages (Franzen et al. 1998; Rabchevsky and
Streit 1997; Lazarov-Spiegler et al. 1996) or the neutral-
ization of myelin-associated growth inhibitors (Schnell
et al. 1994; Schnell and Schwab 1990). Most of these ap-
proaches succeeded in stimulating axon growth but failed
to elongate the lesioned axons within their appropriate
pathway. However, a topographically correct regeneration
is a prerequisite for the reestablishment of original topo-
graphic reinnervation, and the more precise the reinnerva-
tion, the greater the functional improvements will be.
Thus, it emerges that a crucial and outstanding problem
of regeneration-promoting strategies is the accuracy rath-
er than the vigor of regeneration.
The lesion scar seems to be a major barrier that dis-
turbs the required continuous pathway and prevents a to-
pographic reconstruction of a lesioned pathway. Several
decades ago Windle and colleagues (Windle et al. 1951)
made the pioneering studies of injecting a pyrogenic bac-
terial polysaccharide, pyromen, into the lesioned spinal
cords of cats. They reported both reduction of astrogliosis
and enhanced neuritic regeneration. Despite the fact that
the regeneration was not permanent, they provided the
first evidence that the scar constitutes a barrier to regrow-
ing axons. Subsequent studies have applied a variety of
other manipulations, to interfere with the physiology of
the reactive astrocyte. These include surgical removal of
the scar, administration of hormones (Puchala and Windle
1977) and more recent strategies such as X-irradiation
(Kalderon and Fuks 1996) and the application of gluco-
corticoids (Li and David 1996) or antibodies to transform-
ing growth factor b (TGFb) (Logan et al. 1994) (Table 2).
Only single of these approaches improved the regenera-
tion outcome, but they failed to allow regeneration within
the normal pathway, which is a prerequisite for topo-
graphic and coordinate synaptic reinnervation.
A second series of experiments was performed to alter
the texture or density of the BM. In these early efforts en-
zymes such as collagenase, trypsin or elastase were inject-
ed into a damaged region (Freeman 1952; Puchala and
Windle 1977; Guth et al. 1980) to degrade the BM (Ta-
ble 2). Unfortunately, the unavoidable, additional attack
of the BM of blood vessels led to severe changes in vas-
cular supply and limited the regeneration success. Anoth-
er more indirect way to modify the nature of the BM is the
implantation of glial cells, such as astrocytes or Schwann
cells (Emmett et al. 1988; Smith and Miller 1991;
Wunderlich et al. 1994; Stichel et al. 1996; Raisman et
al. 1993; Harvey et al. 1995; Martin et al. 1991; Brook
et al. 1994) (Table 2). In our own work we have used
the minimal invasive microtransplantation approach to in-
troduce a Schwann cell suspension into the transected
postcommissural fornix (Stichel et al. 1996). We were
able to show that the injected Schwann cells become inte-
grated into the neuropil and promote structural recovery
of the projection. The implanted cells seem to interfere
with the scarring process by (a) establishing a guiding
framework, (b) disaggregating the dense cellular network
of the scar and/or (c) physically preventing the infiltration
of scar-promoting cells. In line with the latter effect,
Campbell and coworkers (Galuske et al. 1996) tended to
protect the lesion site from encroachment by collagenous
scar tissue by wrapping the severed ends of the spinal
cord into porous material. The porous material became
calcified and thereby impermeable to regrowing axons.
Recently, we developed a new experimental strategy to
reduce the lesion-induced deposition of the BM. A local
injection of either antibodies against Coll IV or 2, 2di-
pyridyl (DPY), an inhibitor of collagen triple helix forma-
tion and synthesis, interfered with the posttranslational
processing of the major BM component, Coll IV. This
treatment allowed massive axonal regeneration across
the former impermeable lesion scar (Fig. 1C,D). Regrow-
ing fornix axons followed their former pathway, reinner-
vated their normal target, were remyelinated and recov-
Table 2 Experimental strategies to overcome the lesion scar
Excision of scar
X-irradiation
Implantation: Porous material
Glial cells
Infusion/injection: Pyrogenic bacterial polysaccharide
(pyromen)
Trypsin, elastase, collagenase
Glucocorticoids
Antibodies against transforming growth
factor b
Antibodies against collagen IV
Prolylhydroxylase inhibitor
Hormones
7
ered normal conduction properties (Stichel et al. 1997,
1998a). The application of collagen-reducing agents did
not affect the quantity of vascular BM in the surrounding
neuropil (Stichel et al. 1998a) nor did the BM reduction
influence the distribution pattern and density of reactive
astrocytes or microglial cells within the scar and the peri-
lesion area (Stichel et al. 1998b).
However, up to now the nature of the inhibitory mech-
anisms mediated by the BM is still a matter of specula-
tion. Since BM might possess many holes and fenestra-
tions with a diameter of 0.55 m (Komuro 1985; Warfel
and Hull 1988), it is unlikely that the BM represents a
physical barrier for growing axons. Another potential
source of BM-mediated growth inhibition could be repul-
sive molecules associated with the membrane matrix. In
support of this assumption several reports show that
growth-inhibitory proteoglycans, such as chondroitin sul-
fate and heparan sulfate proteoglycans (Ard and Bunge
1988; Snow et al. 1990; Snow et al. 1991), are major con-
stituents of the BM (Timpl and Dziadek 1986; Stichel et
al. 1998b; Couchman et al. 1984). On the other hand, it is
apparent that the myelin-associated proteins NI35/250
and MAG are not involved in the formation of such an in-
hospitable territory. Evidence for this conclusion is (a) ab-
sence of these molecules from the scar tissue (Fig. 1C)
(Stichel et al. 1995b), (b) failure of neutralizing approach-
es to stimulate axonal regeneration across the scar tissue
(Schnell and Schwab 1990, 1993) and (c) the fact that my-
elin or myelin debris beyond the lesion scar can be highly
permissive for growing axons (Davies et al. 1997; Stichel
et al. 1995b).
Taken together, recent data provide strong evidence
that the lesion-induced BM itself is the primary determi-
nant of scar impermeability and hence a major constraint
to axonal regeneration. Exactly what aspect of this matrix
comprises the regeneration barrier is not yet defined. In
the future it will be of crucial importance to characterize
the molecular composition of the BM in CNS scar and to
define the role of its components in the process of axonal
regeneration.
References
Ajemian A, Ness R, David S (1994) Tenascin in the injured rat optic
nerve and in non-neuronal cells in vitro: potential role in neural
repair. J Comp Neurol 340:233242
Alonso G, Privat A (1993a) Neuropeptide Y-producing neurons of
the arcuate nucleus regenerate axons after surgical deafferenta-
tion of the mediobasal hypothalamus. J Neurosci Res 3:215234
Alonso G, Privat A (1993b) Reactive astrocytes involved in the for-
mation of lesional scars differ in the mediobasal hypothalamus
and in other forebrain regions. J Neurosci Res 34:523538
Ard MD, Bunge RP (1988) Heparan sulfate proteoglycan and lam-
inin immunoreactivity on cultured astrocytes: relationship to dif-
ferentiation and neurite outgrowth. J Neurosci 8:28442858
Aubert I, Ridet J-L, Gage FH (1995) Regeneration in the adult mam-
malian CNS: guided by development. Curr Opin Neurobiol
5:625635
Auger MJ, Ross JA (1992) The biology of the macrophage. In: Lew-
is CE, McGee JOD (eds) The macrophage. Oxford University
Press, Oxford, pp 174
Bartsch S, Bartsch U, Drries U, Faissner A, Weller A, Ekblom P,
Schachner M (1992) Expression of tenascin in the developing
and adult cerebellar cortex. J Neurosci 12:736749
Bernstein JJ, Getz R, Jefferson M, Kelemen M (1985) Astrocytes se-
crete basal lamina after hemisection of rat spinal cord. Brain Res
327:135141
Berry M, Maxwell WL, Mathewson A, McConnell P, Ashhurst DE,
Thomas GE (1983) Deposition of scar tissue in the central ner-
vous system. Acta Neurochir Suppl 32:3153
Blaugrund E, Lavie V, Cohen J, Solomon A, Schreyer DJ, Schwartz
M (1993) Axonal regeneration is associated with glial migra-
tion: comparison between the injured optic nerves of fish and
rats. J Comp Neurol 330:105112
Blight AR (1991) Morphometric analysis of blood vessels in chronic
experimental spinal cord injury: hypervascularity and recovery
of function. J Neurol Sci 106:158174
Bovolenta P, Wandosell F, Nieto-Sampedro M (1992) CNS glial
scar tissue: a source of molecules which inhibit central neurite
outgrowth. Prog Brain Res 94:367379
Bregman BS, McAtee M, Dai HN, Kuhn PL (1997) Neurotrophic
factors increase axonal growth after spinal cord injury and trans-
plantation in the adult rat. Exp Neurol 148:475494
Brodkey JA, Gates MA, Laywell ED, Steindler DA (1993) The com-
plex nature of interactive neuroregeneration-related molecules.
Exp Neurol 123:251270
Brook GA, Lawrence JM, Shah B, Raisman G (1994) Extrusion
transplantation of Schwann cells into the adult rat thalamus in-
duces directional host axon growth. Exp Neurol 126:3143
Canning DR, Hke A, Malemud CJ, Silver J (1996) A potent inhib-
itor of neurite outgrowth that predominates in the extracellular
matrix of reactive astrocytes. Int J Dev Neurosci 14:153175
Carbonetto S (1984) The extracellular matrix of the nervous system.
Trends Neurosci 10:382387
Chamak B, Morandi V, Mallat M (1994) Brain macrophages stimu-
late neurite growth and regeneration by secreting thrombospon-
din. J Neurosci Res 38:221233
Cheng H, Cao Y, Olson L (1996) Spinal cord repair in adult paraple-
gic rats: partial restoration of hind limb function. Science
273:510513
Choi BH (1997) Role of the basement membrane in neurogenesis
and repair of injury in the central nervous system. Microsc
Res Tech
Clemente CD (1955) Structural regeneration in the mammalian cen-
tral nervous system and the role of neuroglia and connective tis-
sue. In: Windle WF (ed) Regeneration in the nervous system.
Charles C. Thomas, Springfield, Ill, pp 147161
Couchman JR, Caterson B, Christner JE, Baker JR (1984) Mapping
by monoclonal antibody detection of glycosaminoglycans in
connective tissues. Nature 307:650652
David S, Ness R (1993) Heterogeneity of reactive astrocytes. In:
Fedoroff S (ed) Biology and pathology of astrocyte-neuron in-
teractions. Plenum Press, New York, pp 303312
Davies SJA, Fitch MT, Memberg SP, Hall AK, Raisman G, Silver J
(1997) Regeneration of adult axons in white matter tracts of the
central nervous system. Nature 390:680683
Eddleston M, Mucke L (1993) Molecular profile of reactive astro-
cytes implications for their role in neurologic disease. Neuro-
science 54:1536
Elkabes S, DiCicco-Bloom EM, Black IB (1996) Brain microglia/
macrophages express neurotrophins that selectively regulate
microglial proliferation and function. J Neurosci 16:2508
2521
Emmett CJ, Lawrence JM, Seeley PL, Raisman G (1988) Studies of
the behaviour of purified rat astrocytes after transplantation into
syngeneic adult brain. In: Gash DM, Sladek JR (eds) Elsevier
Science, Amsterdam, pp 383386
Faissner A (1997) Glial derived extracellular matrix components:
important roles in axon growth and guidance. Neuroscientist
3:371380
Fawcett J (1997) Astrocytic and neuronal factors affecting axon re-
generation in the damaged central nervous system. Cell Tissue
Res 290:371377
8
Feringa ER, Kowalski TF, Vahlsing HL (1980) Basal lamina forma-
tion at the site of spinal cord transection. Ann Neurol 8:148154
Fernaud-Espinosa I, Nieto-Sampedro N, Bovolenta P (1993) Differ-
ential activation of microglia and astrocytes in aniso- and iso-
morphic gliotic tissue. Glia 8:277291
Frank M, Wolburg H (1996) Cellular reactions at the lesion site after
crushing of the rat optic nerve. Glia 16:227240
Franzen R, Schoenen J, Leprince P, Joosten EA, Moonen G, Martin
D (1998) Effects of macrophage transplantation in the injured
adult rat spinal cord: a combined immunocytochemical and bio-
chemical study. J Neurosci Res 51:316327
Freeman LW (1952) Return of function after complete transection of
the spinal cord of the rat, cat and dog. Ann Surg 136:193205
Frisn J, Fried K, Sjgren AM, Risling M (1993) Growth of ascend-
ing spinal axons in CNS scar tissue. Int J Dev Neurosci 4:461
475
Galuske RAW, Kim D-S, Castrn E, Thoenen H, Singer W (1996)
Brain-derived neurotrophic factor reverses experience-depen-
dent synaptic modifications in kitten visual cortex. Eur J Neuro-
sci 8:15541559
Geisert EE, Williams GS, Bidanset DJ (1992) A CNS specific pro-
teoglycan associated with astrocytes in rat optic nerve. Brain
Res 571:165168
Grierson JP, Petroski RE, Ling DSF, Geller HM (1990) Astrocyte to-
pography and tenascin/cytotactin expression: correlation with the
ability to support neurite outgrowth. Dev Brain Res 55:1119
Guth L, Albuquerque EX, Deshpande SS, Barrett CP, Donati EJ,
Warnick JE (1980) Ineffectiveness of enzyme therapy on regen-
eration in the transected spinal cord of the rat. J Neurosurg
52:7386
Guth L, Reier PJ, Barrett CP, Donati EJ (1983) Repair of the mam-
malian spinal cord. Trends Neurosci 6:120124
Hall DE, Reichardt LF, Crowley E, Holley B, Moezzi H, Sonnen-
berg A, Damsky CH (1990) The a1/b1 and a6/b1 integrin het-
erodimers mediate cell attachment to distinct sites of laminin.
J Cell Biol 110:21752184
Harvey AR, Plant GW, Tan MML (1995) Schwann cells and the re-
growth of axons in the mammalian CNS: a review of transplan-
tation studies in the rat visual system. Clin Exp Pharmacol Phy-
siol 22:569579
Hatten ME, Liem RKH, Shelanski ML, Mason CA (1991) Astroglia
in CNS injury. Glia 4:233243
Houle JD, Ye JH, Kane DJ (1996) Axonal regeneration by chroni-
cally injured supraspinal neurons can be enhanced by exposure
to insulin-like growth, basic fibroblast growth factor or trans-
forming growth factor beta. Rest Neurol Neurosci 10:205215
Hke A, Silver J (1996) Proteoglycans and other repulsive mole-
cules in glial boundaries during development and regeneration
of the nervous system. Prog Brain Res 108:149163
Jaeger CB, Blight AR (1997) Spinal compression injury in guinea
pigs: structural changes of endothelium and its perivascular cell
associations after blood-brain barrier breakdown and repair. Exp
Neurol 144:381399
Joosten LS (1996) Integrin's possible functions in the adult CNS.
Trends Neurosci 19:6872
Kalaria RN, Pax AB (1995) Increased collagen content of cerebral
microvessels in Alzheimer's disease. Brain Res 705:349352
Kalderon N, Fuks Z (1996) Structural recovery in lesioned adult
mammalian spinal cord by X-irradiation of the lesion site. Proc
Natl Acad Sci 93:1117911184
Klagsbrun M, D'Amore PA (1991) Regulators of angiogenesis. Ann
Rev Physiol 53:217239
Komuro T (1985) Fenestrations of the basal lamina of intestinal villi
of the rat. Scanning and transmission microscopy. Cell Tissue
Res 239:183188
Kramer RH, Marks N (1989) Identification of integrin collagen re-
ceptors on human melanoma cells. J Biol Chem 264:46844688
Krger S, Sievers J, Hansen C, Sadler M, Berry M (1986) Three
morphologically distinct types of interface develop between
adult host and fetal brain transplants: implications for scar for-
mation in the adult central nervous system. J Comp Neurol
249:103116
Landis DMD (1994) The early reactions of non-neuronal cells to
brain injury. Ann Rev Neurosci 17:133151
Lazarov-Spiegler O, Solomon AS, Zeev-Brann AB, Hirschberg DL,
Lavie V, Schwartz M (1996) Transplantation of activated mac-
rophages overcomes central nervous system regrowth failure.
FASEB J 10:12961302
Li MS, David S (1996) Topical glucocorticoids modulate the lesion
interface after cerebral cortical stab wounds in adult rats. Glia
18:306318
Li Y, Raisman G (1995) Sprouts from cut corticospinal axons persist
in the presence of astrocytic scarring in long-term lesions of the
adult rat spinal cord. Exp Neurol 134:102111
Li Y, Filed PM, Raisman G (1997) Repair of adult rat corticospinal
tract by transplants of olfactory ensheathing cells. Science
277:20002002
Lindsay RM (1986) Reactive gliosis. In: Fedoroff S, Vernadakis A
(eds) Astrocytes. Academic Press, Orlando, pp 231262
Lips K, Stichel CC, Mller HW (1995) Restricted appearance of ten-
ascin and chondroitin sulfate proteoglycans after transection and
sprouting of adult rat postcommissural fornix. J Neurocytol
24:449464
Logan A, Berry M, Gonzalez AM, Frautschy SA, Spron MB, Baird
A (1994) Effects of transforming growth factor b1 on scar pro-
duction in the injured central nervous system of the rat. Eur J
Neurosci 6:355363
Martin D, Schoenen J, Delre P, Leprince P, Rogister B, Moonen G
(1991) Grafts of syngeneic cultured, adult dorsal root ganglion-
derived Schwann cells to the injured spinal cord of adult rats:
preliminary morphological studies. Neurosci Lett 124:4448
Maxwell WL, Duance VC, Lehto M, Ashhurst DE, Berry M (1984)
The distribution of types I, III, IV and V collagens in penetrant
lesions of the central nervous system of the rat. Histochem J
16:12191229
Maxwell WL, Follows R, Ashhurst DE, Berry M (1990) The re-
sponse of the cerebral hemisphere of the rat to injury. I. The ma-
ture rat. Philos Trans R Soc Lond B 328:36500
McLoon SC (1986) Response of astrocytes in the visual system to
Wallerian degeneration: an immunohistochemical analysis of
laminin and glial fibrillary acidic protein (GFAP). Exp Neurol
91:613621
Mller HW, Junghans U, Kappler J (1996) Astroglial neurotrophic
and neurite-promoting factors. In: Bell C (ed) Chemical factors
in neural growth, degeneration and repair. Elsevier Science,
Amsterdam, pp 377397
Nicholson C, Sykova E (1998) Extracellular space structure revealed
by diffusion analysis. Trends Neurosci 21:207214
Paino CL, Fernandez-Valle C, Bates ML, Bunge MB (1994) Re-
growth of axons in lesioned adult rat spinal cord: promotion
by implants of cultured Schwann cells. J Neurocytol 23:433452
Paulsson M (1992) Basement membrane proteins: structure assem-
bly, and cellular interactions. Crit Rev Biochem Mol Biol
27:93127
Perry VH, Gordon S (1988) Macrophages and microglia in the ner-
vous system. Trends Neurosci 11:273277
Puchala E, Windle WF (1977) The possibility of structural and func-
tional restitution after spinal cord injury. A review. Exp Neurol
55:142
Puckett WR, Hiester ED, Norenberg MD, Marcillo AE, Bunge RP
(1997) The astroglial response to Wallerian degeneration after
spinal cord injury in humans. Exp Neurol 148:424432
Rabchevsky AG, Streit WJ (1997) Grafting of cultured microglial
cells into the lesioned spinal cord of adult rats enhances neurite
outgrowth. J Neurosci Res 47:3448
Raisman G, Lawrence JM, Brook GA (1993) Schwann cells trans-
planted into the CNS. Int J Dev Neurosci 11:651669
Ramon-Cuto A, Nieto-Sampedro N (1994) Regeneration into the
spinal cord of transected dorsal root axons is promoted by en-
sheathing glia transplants. Exp Neurol 127:232244
Ramon-Cuto A, Plant GW, Avila J, Bunge MB (1998) Long-dis-
tance axonal regeneration in the transected adult rat spinal cord
is promoted by olfactory ensheathing glia transplants. J Neurosci
18:38033815
9
Ramn y Cajal S (1928) Degeneration and regeneration of the ner-
vous system. Hafner, New York
Reier PJ, Stensaas LJ, Guth L (1983) The astrocytic scar as an im-
pediment to regeneration in the central nervous system. In: Kao
CC, Bunge RP, Reier PJ (eds) Spinal cord reconstruction. Raven
Press, New York, pp 163195
Richardson PM, Issa VMK, Shemie S (1982) Regeneration and ret-
rograde degeneration of axons in the rat optic nerve. J Neurocy-
tol 11:949966
Ridet J-L, Malhotra SK, Privat A, Gage FH (1997) Reactive astro-
cytes: cellular and molecular cues to biological function. Trends
Neurosci 20:570577
Rutka JT, Apodaca G, Stern R, Rosenblum M (1988) The extracel-
lular matrix of the central and peripheral nervous systems: struc-
ture and function. J Neurosurg 69:155170
Schnell L, Schwab ME (1990) Axonal regeneration in the rat spinal
cord produced by an antibody against myelin-associated neurite
growth inhibitors. Nature 343:269272
Schnell L, Schwab ME (1993) Sprouting and regeneration of lesion-
ed corticospinal tract fibres in the adult rat spinal cord. Eur J
Neurosci 5:11561171
Schnell L, Schneider R, Kolbeck R, Barde Y-A, Schwab ME (1994)
Neurotrophin-3 enhances sprouting of corticospinal tract during
development and after adult spinal cord lesion. Nature 367:170
173
Sivron T, Cohen A, Duvdevani R, Jeserich G, Schwartz M (1990)
Glial response to axonal injury: in vitro manifestation and impli-
cation for regeneration. Glia 3:267276
Smith GM, Miller RH (1991) Immature type-1 astrocytes suppress
glial scar formation, are motile and interact with blood vessels.
Brain Res 543:111122
Snow AD, Lemmon V, Carrino DA, Caplan AI, Silver J (1990) Sul-
fated proteoglycans in astroglial barriers inhibit neurite out-
growth in vitro. Exp Neurol 109:111130
Snow DM, Watanabe M, Letourneau PC, Silver J (1991) A chondro-
itin sulfate proteoglycan may influence the direction of retinal
ganglion cell outgrowth. Development 113:14731485
Stichel-Gunkel CC (1997) The role of microenvironment in axonal
regeneration. Springer, Berlin Heidelberg New York (Advances
in anatomy, embryology and cell biology, vol 137)
Stichel CC, Mller HW (1994a) Extensive and long-lasting changes
of glial cells following transection of the postcommissural for-
nix in the adult rat. Glia 10:89100
Stichel CC, Mller HW (1994b) Relationship between injury-in-
duced astrogliosis, laminin expression and axonal sprouting in
the adult rat brain. J Neurocytol 23:615630
Stichel CC, Kappler J, Junghans U, Koops A, Kresse H, Mller HW
(1995a) Differential expression of the small chondroitin/derma-
tan sulfate proteoglycans decorin and biglycan after injury of the
adult rat brain. Brain Res 704:263274
Stichel CC, Wunderlich G, Schwab ME, Mller HW (1995b) Clear-
ance of myelin constituents and axonal sprouting in the transec-
ted postcommissural fornix of the adult rat. Eur J Neurosci
7:401411
Stichel CC, Lips K, Wunderlich G, Mller HW (1996) Reconstruc-
tion of transected postcommissural fornix in adult rat by
Schwann cell suspension grafts. Exp Neurol 140:2136
Stichel CC, Hermanns S, Luhmann H, Mller HW (1997) Regener-
ation of lesioned axons across CNS scar tissue following modu-
lation of the lesion interface. Neurotrauma Symp (Abstr)
Stichel CC, Hermanns S, Lausberg F, Niermann H, Luhmann H,
Mller HW (1998a) Massive regeneration of lesioned CNS ax-
ons across scar tissue following modulation of the lesion inter-
face. Eur J Neurosci Suppl 10:25.01 (Abstr)
Stichel CC, Niermann H, Hermanns S, Lausberg F, D'Urso D, Ml-
ler HW (1998b) Axon growth through the modified CNS scar.
Eur J Neurosci Suppl 10:25.03 (Abstr)
Streit WJ, Graeber MB, Kreutzberg GW (1988) Functional plasticity
of microglia: a review. Glia 1:301307
Timpl R, Dziadek M (1986) Structure, development and molecular
pathology of basement membranes. Int Rev Exp Pathol 29:1
112
Tuszynski MH, Gabriel K, Gage FH, Suhr S, Meyer S, Rosetti A
(1996) Nerve growth factor delivery by gene transfer induces
differential outgrowth of sensory, motor, and noradrenergic neu-
rites after adult spinal cord injury. Exp Neurol 137:157173
Vidal-Sanz M, Bray GM, Villegas-Prez MP, Thanos S, Aguayo AJ
(1987) Axonal regeneration and synapse formation in the supe-
rior colliculus by retinal ganglion cells in the adult rat. J Neuro-
sci 7:28942909
Warfel K, Hull MT (1988) Basal lamina fenestrations in the human
colon: transmission and scanning electron microscope study.
Anat Rec 220:6875
Windle WF, Clemente CD, Chambers WW (1951) Regeneration in
the spinal cord of the cat and dog. Arch Neurol Psychiatry
65:261262
Wunderlich G, Stichel CC, Schroeder WO, Mller HW (1994)
Transplants of immature astrocytes promote axonal regeneration
in the adult rat brain. Glia 10:4958
Xu XM, Gunard V, Kleitman N, Bunge MB (1995) Axonal regen-
eration into Schwann cell-seeded guidance channels grafted into
transected adult rat spinal cord. J Comp Neurol 351:145160
Yurchenco PD, O'Rear JJ (1994) Basal lamina assembly. Curr Opin
Cell Biol 6:674681
10