MICROSCOPY RESEARCH AND TECHNIQUE 41:431–440 (1998)

Galactolipids in the Formation and Function

of the Myelin Sheath
JEFFREY L. DUPREE,1 KINUKO SUZUKI,1,2 AND BRIAN POPKO1,3,4*
1UNC Neuroscience Center, Univeristy of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599
2Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599
3Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599
4Program in Molecular Biology and Biotechnology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599

KEY WORDS galactocerebroside; sulfatide; UDP-galactose:ceramide galactosyltransferase;

ultrastructure
ABSTRACT Among the most abundant components of myelin are the galactolipids galactocere-
broside (GalC) and sulfatide. In spite of this abundance, the roles that these molecules play in the
myelin sheath are not well understood. Until recently, our concept of GalC and sulfatide functions
had been principally defined by immunological and chemical perturbation studies that implicate
these lipids in oligodendrocyte differentiation, myelin formation, and myelin stability. Recently,
however, genetic studies have allowed us to re-analyze the functions of these lipids. Two laboratories
have independently generated mice that are incapable of synthesizing either GalC or sulfatide by
inactivating the gene encoding the enzyme UDP-galactose:ceramide galactosyltransferase (CGT),
which is required for myelin galactolipid synthesis. These galactolipid-deficient animals exhibit a
severe tremor, hindlimb paralysis, and display electrophysiological deficits in both the central and
peripheral nervous systems. In addition, ultrastructural studies have revealed hypomyelinated
white matter tracts with unstable myelin sheaths and a variety of myelin abnormalities including
altered node length, reversed lateral loops, and compromised axo-oligodendrocytic junctions.
Collectively, these observations indicate that cell-cell interactions, which are essential in the
formation and maintenance of a properly functioning myelin sheath, are compromised in these
galactolipid-deficient mice. Microsc. Res. Tech. 41:431–440, 1998. r 1998 Wiley-Liss, Inc.

INTRODUCTION understanding of the role that these galactolipids play

The dry weight of myelin is approximately 70% lipid,
with the galactolipids, galactocerebroside (GalC) and
its sulfated derivative sulfatide, comprising almost
one-third of the lipid mass of this highly specialized
membranous sheath (reviewed by Norton and Cammer,
1984). Because of this abundance, combined with the
fact that GalC is the first myelin component to appear
on myelinating cells (Pfeiffer et al., 1993), GalC and
sulfatide have been implicated in a variety of roles
essential for the establishment and maintenance of a
properly functioning myelin sheath (reviewed by Coet-
zee et al., 1998; Stoffel and Bosio, 1997). In order to
explore such possible functions, a number of investiga-
tors have employed galactolipid perturbation tech-
niques both in vitro and in vivo, and have identified
several potential functions of these lipids with regard to
myelin. In addition to these perturbation studies, func-
tions of GalC and sulfatide have been investigated
genetically. Recently, mice have been generated with a
mutation in the gene that encodes UDP-galactose:
ceramide galactosyltransferase (CGT) (Coetzee et al.,
1996; Bosio et al., 1996), the enzyme that catalyzes the
final step in the synthesis of GalC (Fig. 1) (Morell and
Radin, 1969). The mutant mice, which are incapable of
synthesizing either GalC or sulfatide, displayed a vari-
ety of deficits in myelin structure, function, and stabil-
ity. Detailed biochemical and ultrastructural examina-
tions of these mutants have greatly increased our
in myelinating cell differentiation and function.
GALACTOLIPID PERTURBATION STUDIES
Over the past 2 decades, perturbation strategies have
been used to investigate the possible roles of galactolip-
ids in the establishment of a properly functioning
myelin sheath. In these studies, many different antibod-
ies have been used in a variety of in vitro and in vivo
systems, and the findings from these reports have
implicated GalC and sulfatide in oligodendrocytic differ-
entiation, myelin formation, and myelin stability (re-
viewed in Benjamins and Dyer, 1990; Dyer, 1993).
Oligodendrocyte Differentiation
Both immunological and chemical perturbations of
GalC and sulfatide have dramatically altered the matu-
ration process in myelin-forming cells. Following treat-
ment with an antibody that recognizes both sulfatide
and GalC (Ranscht monoclonal antibody (R-mAb);
Ranscht et al., 1982), and possibly other uncharacter-
ized oligodendrocyte markers (Bansal and Pfeiffer,
1989), cultured embryonic oligodendrocytes fail to ter-
Contract grant sponsor: NIH; Contract grant numbers: NS27336, NS01637;
Contract grant sponsor: National Multiple Sclerosis Society.
*Correspondence to: Dr. Brian Popko, UNC Neuroscience Center, CB# 7250,
University of North Carolina, Chapel Hill, NC 27599–7250.
E-mail: bpopko@css.unc.edu
Received 19 November 1997; Accepted in revised form 12 January 1998

r 1998 WILEY-LISS, INC.


432 J.L. DUPREE ET AL.
Fig. 1. Galactocerebroside and glucocerebroside are synthesized
from the common precursor ceramide. In the galactolipid deficient
mice, the absence of CGT results in an abundance of GlcC, a molecule
that is usually present in very low quantities since it is used as an
minally differentiate at the RNA level (Bansal and
Pfeiffer, 1989). Upon removal of the antibody, the cells
rapidly resume differentiation and present a morphol-
ogy that is consistent with mature oligodendrocytes.
From these observations, the authors propose that
binding of the R-mAb blocks differentiation by inhibit-
ing the function of GalC and sulfatide. Since treatment
of these cells with the 01 mAb, a galactolipid antibody
that does not recognize sulfatide (Bansal et al., 1989), is
ineffective at blocking differentiation, Bansal and
Pfeiffer (1989) postulate that sulfatide is the target
molecule responsible for oligodendrocyte differentia-
tion. Nevertheless, the inhibition of sulfation (i.e.,
reduced sulfatide synthesis) does not block oligodendro-
cyte differentiation (Bansal and Pfeiffer, 1994a), suggest-
ing that sulfatide is not essential for the maturation of
oligodendrocytes. Furthermore, the addition of a sul-
fatide-directed antibody to cultured oligodendrocytes
stimulates differentiation as assessed by the expression
of myelin specific proteins (Bansal et al., 1988). The
authors propose that the sulfatide antibody stimulates
a sulfatide-mediated cascade of intracellular events
that facilitates myelin gene expression. In addition to
the possible involvement of sulfatide, GalC may also
play a role in the development of these cells. Anti-GalC
serum induces cytoskeletal and morphological changes
in cultured oligodendrocytes, therefore suggesting that
GalC may mediate maturation through transmem-
brane signaling (Dyer and Benjamins, 1988, 1990). In
total, these in vitro perturbation studies are somewhat
contradictory and confusing, leaving open the question
of what role the myelin galactolipids play in oligodendro-
cyte differentiation.
Myelin Formation
Numerous antibody perturbation studies also report
that antisera against cerebroside inhibits in vitro my-
elination (Dorfman et al., 1976, 1978, 1979; Dubois-
Dalcq et al., 1970; Fry et al., 1974; Hruby et al., 1977;
Raine et al., 1978). The addition of the R-mAb to
peripheral nervous system (PNS) co-cultures does not
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intermediate in the synthesis of gangliosides. CGT: UDP-galactose:
ceramide galactosyltransferase; GlcT-1: UDP-glucose ceramide:
glucosyltransferase.
interfere with the initial interaction between the axon
and the Schwann cell but inhibits the process of mes-
axon wrapping (Ranscht et al., 1987; Owens and Bunge,
1990). Oligodendrocytes expressing terminal differen-
tiation markers retract cellular processes and mem-
brane sheets in the presence of a GalC directed anti-
body (Bansal and Pfeiffer, 1994b). Moreover, the
implantation of anti-galactolipid producing hybridoma
cells into the spinal cords of immature rats either
completely disrupts myelin formation (Rosenbluth et
al., 1994) or produces myelin sheaths with widely
separated lateral loops and increased lamellae spacing
(Rosenbluth et al., 1995). Taken together, these observa-
tions indicate that galactolipids are essential in the
process of myelinogenesis. More specifically, separated
lateral loops and abnormally spaced lamellae suggest
that these lipids mediate membrane-membrane interac-
tions that are critical for proper myelin formation.
Myelin Stability
In addition to their possible involvement in oligoden-
drocyte differentiation and myelin formation, antibody
treatment studies provide morphological evidence that
galactolipids are essential in maintaining myelin struc-
ture. Myelinated spinal cord slice cultures treated with
cerebroside antibody yield myelin fragmentation and a
loss of sheath integrity (Fry et al., 1974; Saito et al.,
1986). Similarly, the addition of anti-GalC serum pro-
duces intramyelinic splitting, ‘‘smudged’’ myelin and
oligodendrocyte degeneration in cerebellar slice cul-
tures (Saida et al., 1979b) and myelin collapse in spinal
cord explants (Roth et al., 1985). The injection of GalC
antibodies into adult animals induces segmental demy-
elination with interlamellar separation and myelin
splitting in the PNS (Saida et al., 1979a) and myelin
vesiculation leading to complete demyelination in the
central nervous system (CNS) (Sergott et al., 1986). In
mature rats with implanted anti-GalC producing hy-
bridoma cells, axonal processes in the vicinity of the
implants frequently lack myelin, indicating that these
fibers are demyelinated (Rosenbluth et al., 1995). In

MYELIN GALACTOLIPIDS
Fig. 2. The life expectancy of the CGT-deficient mice is dramatically decreased with few animals
surviving beyond PND 90.
light of the findings of Saito et al. (1986), who demon-
strated that surface GalC was internalized following
interaction with the antibody, demyelination may be
the result of insufficient galactolipid-mediated cell-cell
interactions. Collectively, these studies suggest that
GalC is essential in maintaining the myelin sheath.
Although the antibody perturbation studies indicate
that the galactolipids are involved in oligodendrocyte
differentiation, myelin formation, and myelin stability,
several caveats must be addressed to properly interpret
these data. Only a few of the studies demonstrate that
the loss of cellular differentiation and myelin structure
is independent of complement-mediated events (Bansal
and Pfeiffer, 1989; Dorfman et al., 1979; Fry et al.,
1974). Secondly, the antibodies used in these reports
display considerable cross reactivity to various lipids,
making it difficult to attribute their effect to a specific
molecule (Bansal et al., 1989, 1992; Bansal and Pfeiffer,
1992). Finally, reports rarely demonstrate the mecha-
nism responsible for the effects resulting from antisera
binding to the galactolipid target. Therefore, several
questions critical to the understanding of the models
are left unanswered: (1) Do the antibodies used in these
studies inhibit or stimulate GalC/sulfatide-mediated
events? (2) Is the antibody/antigen complex internal-
ized, effectively reducing the number of functional
antigen molecules on the cell surface as demonstrated
by Saito et al. (1986) and Ranscht et al. (1987)? (3) Are
the functions of neighboring molecules influenced by
the formation of the antibody/antigen complex? There-
fore, the conclusions presented in these perturbation
studies must be tempered with these reservations
making accurate assessment of the data difficult.
CGT-DEFICIENT MICE
The recent isolation of the gene that encodes CGT
(Schulte and Stoffel, 1993; Stahl et al., 1994; Schaeren-
Wiemers et al., 1995), the enzyme that catalyzes the
final step in the synthesis of GalC (Morell and Radin,
1969), has allowed for the genetic analysis of myelin
galactolipid function. Mice with a disruption in the
CGT gene were generated, and the success of these
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433
methods has been demonstrated by Northern blot
analyses, thin layer chromatography (Bosio et al., 1996;
Coetzee et al., 1996), in situ hybridization, and CGT
activity assays (Bosio et al., 1996). Phenotypically,
these animals demonstrate a pronounced tremor both
at rest and during movement as early as 2 weeks of age,
and their gait is characterized by splayed hindlimbs
with clenched paws and a lowered hindquarter region
with a vertically postured tail. The CGT -/- animals
exhibit a progressive hindlimb paralysis, such that by
postnatal day (PND) 60, many animals are immobile.
The life expectancy of these animals is greatly de-
creased with mutants rarely surviving past PND 90
(Fig. 2) (Coetzee et al., 1996). It is important to point
out that CGT is expressed in tissues other than the
nervous system (Bengtsson et al., 1996; Henseler et al.,
1996; Krivan et al., 1989; Sakakibara et al., 1981;
Seddiki et al., 1996; Shimomura and Kishimoto, 1983;
Ueno et al., 1975; Zalc et al., 1978), and the loss of
function of this enzyme in these non-neural systems
may significantly contribute to the phenotype of these
mutants. For example, preliminary morphological obser-
vations reveal that the kidney and the testis are
dramatically altered in the mutant animals (Dupree et
al., unpublished data).
Oligodendrocyte Differentiation
and Myelin Formation
Although the CGT-deficient mice present a pheno-
type that is consistent with severely altered sheath
structure, these mutants surprisingly display com-
pacted myelin with normal periodicity in both the CNS
and the PNS (Fig. 3) (Bosio et al., 1996; Coetzee et al.,
1996). In addition, RNA and protein levels of myelin
basic protein and proteolipid protein, late stage develop-
mental markers for oligodendrocytes, are comparable
between CGT 2/2 and 1/1 mice at PND 15, 30, 45, and
90 (Coetzee et al., unpublished data; Dupree et al.,
1998). Clearly these findings indicate that, in contrast
to the in vitro perturbation studies, oligodendrocyte
differentiation and myelin formation are not blocked in
the absence of galactolipid function. Thus, galactolipid-
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434 J.L. DUPREE ET AL.

Fig. 3. Myelin sheaths from the ventral columns of the cervical spinal cord of 30-day-old wild-type
mice (A) are thicker than the sheaths from comparable regions in age matched CGT-deficient animals (B).
No difference in sheath thickness was observed in the optic (C,D) or sciatic (E,F) nerves between control
(C and E) and mutant (D and F) mice. Magnification bars 5 1.0 µm.

mediated intracellular signaling is not essential for
oligodendrocyte differentiation.
An unexpected result that both Coetzee et al. (1996)
and Bosio et al. (1996) report is the presence of glucoce-
rebroside (GlcC) in CNS and PNS myelin. GlcC is
usually found only in very low levels in control animals
since it is an intermediate product in ganglioside
biosynthesis (Fig. 1) (Hammarstrom, 1971). In the
absence of CGT activity, the ceramide normally des-
tined for incorporation into GalC is presumably used as
a substrate for GlcC synthesis. Interestingly, some
primitive fish species and certain marine invertebrates
contain GlcC in their myelin (Kishimoto, 1986; Tamai
et al., 1992). These findings, combined with the fact
that GlcC exhibits similar biochemical properties to
GalC (Koynova and Caffrey, 1995), suggest that GlcC
may, at least partially, substitute for GalC function
allowing for the efficient progression of myelin cell
differentiation and myelin formation in the CGT mu-
tant mice. Perhaps in the antibody perturbation studies
described above, the myelinating cells are unable to
compensate for the absence of galactolipids through
GlcC synthesis. When cells are treated with galacto-
lipid antibodies, CGT activity remains, such that ce-
ramide levels are not elevated, preventing increased
GlcC synthesis.
Myelin Structural Abnormalities
Although the CNS myelin sheaths are grossly nor-
mal, they do exhibit several ultrastructural abnormali-
ties. Coetzee et al. (1996) reported that by PND 24, the
dorsal and ventral columns of the spinal cord are

MYELIN GALACTOLIPIDS
Fig. 4. The most frequent aberrant nodal formation observed in the CGT-deficient mice is the
significant increase in the number of heminodes. A high incidence of heminode formation results in long
stretches of unmyelinated axons and may hamper impulse conduction. Magnification bar 5 1.0 µm.
significantly hypomyelinated (Fig. 3A and B) while no
difference in myelin thickness is observed in either the
optic or sciatic nerves (Fig. 3C–F). In addition to
hypomyelination, abnormal nodal and paranodal struc-
tures are commonly observed in the CNS of the CGT
2/2 mice. Nodal length is typically increased and
heminodes, defined as a paranode juxtaposed to an
extended unmyelinated axonal region, are prevalent
(Fig. 4) (Dupree et al., 1998). The lateral loops are
frequently disorganized and face away from the axo-
lemma, and adjacent myelin sheaths with overlapping
paranodal regions that completely occlude node forma-
tion are occasionally observed (Fig. 5) (Dupree et al.,
1998). Since appropriate cell-cell communication is
essential for proper node formation (reviewed in Sch-
erer, 1996; Salzer, 1997), a disruption in these interac-
tions could lead to improper alignment of oligodendro-
cytic processes along the axon. Such misalignments
could explain both lengthened nodes and overlapping
paranodal regions.
Transverse bands, regularly arrayed densities in the
periaxonal space that are prominent features of the
axo-oligodendrocytic paranodal junctional complex, are
absent in the CNS of the CGT-deficient animals (Fig. 6)
(Dupree et al., 1998). The absence of transverse bands
is further indication that myelin galactolipids are essen-
tial in establishing proper cell-cell interactions. Al-
though the exact function of these structures is not
known, they have been implicated in isolating the
periaxonal space from the surrounding extracellular
space, in establishing axolemmal domains and in an-
choring the myelin sheath to the axon (reviewed by
Rosenbluth, 1995). In the absence of these structures,
the lateral loops are not tightly opposed to the axo-
lemma, facilitating the entrance of cellular and acellu-
lar material into the paranodal periaxonal space (Rosen-
bluth, 1987). In support of this hypothesis, Rosenbluth
(1987) observed the intrusion of astrocytic processes
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435
between the myelin sheath and the axolemma in the
myelin deficient rat, a myelin mutant that lacks trans-
verse bands. Similarly, recent data from our laboratory
show that the CGT 2/2 mice also have astrocytic
process penetration into the periaxonal space (Dupree
et al., 1998), an observation that further indicates that
the myelin galactolipids are essential in establishing
proper cell-cell interactions. This hypothesis is further
supported by the observations that CNS lateral loops
frequently face away from the axolemma (Fig. 5), and
occasionally adjacent lateral loops are not closely asso-
ciated and the myelin sheath is not tightly opposed to
the axolemma (Fig. 7). As previously mentioned, widely
spaced lateral loops were also reported in vivo following
the implantation of anti-GalC hybridomas (Rosenbluth
et al., 1995). Tight junctions ordinarily adjoin adjacent
lateral loops, and such abnormal spacing indicates a
disruption in the formation of these junctions. Interest-
ingly, sulfatide has been shown to be a prominent
constituent of the myelin tight junctions known as the
radial component (Karthigasan et al., 1994). Therefore,
tight junction formation between lateral loops and
myelin lamellae may be dependent on the presence of
sulfatide. Taken together, these observations, abnor-
mal node length, absence of transverse bands, reversed
lateral loops, and widely spaced lateral loops, indicate
that GalC and sulfatide are critical components in
establishing proper axo-oligodendrocytic interactions
that ensure proper myelin formation. In contrast to the
CNS, the PNS displays no abnormalities in the struc-
ture of either myelin sheath or the nodes of Ranvier
(Dupree et al., 1998).
Similar ultrastructural myelin abnormalities have
been reported in the CNS of myelin deficient rat
(Rosenbluth, 1987) and shaking pup (Griffiths et al.,
1981). These models, however, are the products of a
point mutation in the proteolipid protein gene that is
lethal to oligodendrocytes (Dentinger et al., 1982; Dun-

436 J.L. DUPREE ET AL.
Fig. 5. Disorganized lateral loops and abnormal node of Ranvier
structure are frequently observed in the CGT-deficient mice. A:
Lateral loop disarray and paranodal regions from adjacent myelin
sheaths (1 and 2) overlap and completely occlude node of Ranvier
can et al., 1983, respectively). In contrast, both biochemi-
cal and ultrastructural data suggest that oligodendro-
cyte viability is not compromised in the CGT-deficient
mice even at PND 90 (Bosio et al., 1996; Dupree et al.,
1998).
Myelin Instability
With age the CGT-deficient mice exhibit extensive
and progressive myelin vacuolation, leading to intrape-
riod line splitting in the ventral columns of the spinal
cord (Fig. 8) (Coetzee et al., 1996). The onset of this
vacuolar degeneration is prior to PND 45, and by PND
90, myelin in the ventral columns of the cervical spinal
cord is virtually absent. This demyelination is likely
related to the absence of transverse bands and altered
lateral loop formation. Since the periaxonal space is
accessible to the extracellular milieu, we propose that
the initial insult that facilitates demyelination occurs
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formation. B: This high-magnification micrograph illustrates lateral
loops that, although organized, are reversed and face away from the
axon (A). Magnification bar 5 0.1 µm.
in the periaxonal space. This space becomes edematous,
causing vacuolar formation between the myelin sheath
and the axolemma (Fig. 6b). Since the lateral loops are
not adjoined by tight junctions, perhaps the increasing
pressure caused by the fluid buildup separates the
axolemma from the sheath and forces the loops and
their corresponding lamella apart, resulting in intrape-
riod line splitting. Such myelin instability in the ab-
sence of functional galactolipids is consistent with the
antibody perturbation studies (Rosenbluth et al., 1995;
Roth et al., 1985; Saida et al., 1979a,b; Saito et al.,
1986; Sergott et al., 1986). Vacuolar myelin degenera-
tion, however, was rarely observed in the dorsal column
of the spinal cord or in the optic nerve and never in the
sciatic nerve. This heterogeneic tolerance to GalC and
sulfatide elimination is not understood but is currently
under investigation.
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MYELIN GALACTOLIPIDS 437

Fig. 6. Transverse bands (arrowheads), regularly spaced densities that are prominent features of the
axo-glial junctional complex, are present in the CGT littermate controls (A) but absent in the
CGT-deficient mice (B). Magnification bar 5 0.1 µm.

Myelin Functional Deficits
Consistent with the structural abnormalities ob-
served in the CNS myelin, electrophysiological deficits
in these mutant have also been reported (Coetzee et al.,
1996; Bosio et al., 1996). Using a double sucrose gap
technique, Coetzee et al. (1996) report that the action
potential amplitude is significantly reduced and the
peak latency is significantly delayed in isolated spinal
cords. The frequency of abnormal node formation and
the prevalence of heminodes likely accounts for the
conduction deficits in the spinal cord of the CGT mutant
animals. In addition, treatment with 4-aminopyridine
(4-AP), a blocker of potassium channels located in the
paranodal region, partially restores the action potential
amplitude in isolated spinal cords from the mutant
animals. Ordinarily, 4-AP has little effect on the action
potentials of myelinated nerve fibers since the potas-
sium channels are occluded by the myelin sheath and
do not participate in the impulse propagation. These
findings indicate that the paranodal regions are acces-
sible to the extracellular milieu, that the tight associa-
tion between the axon and the myelin sheath is dis-
rupted, and that the myelin galactolipids are essential
for the formation of proper axo-oligodendrocytic interac-
tions in the CNS. In contrast, the electrophysiological
deficits observed in the PNS are much less severe,
consistent with the observed normal ultrastructure,
and may be attributable to the changes in the lipid
composition of the sheath (Dupree et al., 1998).
CLOSING REMARKS
In conclusion, the data from both the perturbation
studies and from the structural and functional analyses
of the CGT-deficient mice demonstrate that the galacto-

438 J.L. DUPREE ET AL.
Fig. 7. Paranodal (A) and internodal (B) regions from 30-day-old
CGT-deficient mice. A: The lateral loops, although facing the axon, are
widely spaced and are not joined by tight junctions. In addition a
vacuole, which is observed to the right of the first lateral loop, is
separating the myelin sheath from the axon. Since this axolemma/
lipids are essential in the formation and maintenance
of myelin. One interesting difference between the per-
turbation studies and the analyses of the mutants was
the effect on the PNS. While antibody treatment of the
PNS both in vitro (Owens and Bunge, 1990; Ranscht et
al., 1987) and in vivo (Saida et al., 1979a) resulted in
the disruption of myelin formation and structure, the
findings from the CGT-deficient mice clearly demon-
strate that neither GalC nor sulfatide is required for
the formation of structurally normal PNS myelin. In
addition, the CGT -/- mice conclusively demonstrate
that the galactolipids are not essential for the differen-
tiation of oligodendrocytes since these cells not only
display terminal differentiation markers (Bosio et al.,
1996; Dupree et al., 1998) but also form compact
myelin. Furthermore, the ability of these cells to pro-
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sheath separation precedes any interlamellar splitting, the initial site
of insult appears to be the periaxonal space. B: The low-magnification
electron micrograph better demonstrates this axolemma/sheath sepa-
ration that presumably results in the massive vacuolar degeneration
observed in PND 45 and older animals. Magnification bar 5 0.1 µm.
duce myelin in the absence of such abundant normal
components demonstrates the remarkable compensa-
tory capacity of both the CNS and the PNS.
Although the CGT mutant mice form compacted
myelin, the sheaths in the CNS are regionally thin and
unstable, display a variety of nodal and paranodal
defects, and are functionally impaired. We propose that
these abnormal myelin structures are the result of
disruptions in axo-oligodendrocyte interactions that
are mediated by GalC and sulfatide. Therefore, galacto-
lipids are essential in the formation and maintenance
of a properly functioning myelin sheath. Further study
of the CGT-deficient mice should greatly increase our
understanding of the specific role that these lipids
play with regard to the myelinating cell and myelina-
tion.

MYELIN GALACTOLIPIDS
Fig. 8. A large, presumably edematous vacuole (V) separates the sheath from the axon (A) resulting in
both axolemma/sheath and interlamellar separation of a ventral column nerve fiber from a 45-day-old
CGT-deficient animal. This process of demyelination has previously been named vacuolar degeneration.
Magnification bar 5 0.5 µm.
ACKNOWLEDGMENTS
We thank Dr. Tim Coetzee for providing the data for
Figure 2.
We also thank Dr. Coetzee and Kris Baerwald for
their insightful critique of the manuscript. The work
described in this review from the laboratory of Brian
Popko was supported by a grant from the National
Institute of Health (NIH) (NS27336). B.P. is the recipi-
ent of a Research Career Development Award from the
NIH (NS01637). J.D. is supported by an Advanced
Postdoctoral Fellowship from the National Multiple
Sclerosis Society.
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