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Heparin-binding site in the G domain of the mouse laminin a chain mediates cell attachment and neurite outgrowth
HEPARIN-BINDINGSITE IN THE G DOMAINOF THE MOUSELAMININA CHAINMEDIATES CELL ATTACHMENT AND NEURITEOUTGROWTH KEN-ICHIROTASHIRO” ,MASASHI TAKEICHI’.AND YOSHIHIKOYAMADA’. ‘Department of Medical
School.5-l-l
National
Institute
Nabeshima.Saga.849.Japan. of Dental
Four heparin-binding
*Laboratory
Research.National
domains
Institute
had been
It
is
comprised
of 3084 amino acids.which
several
peptides
from the amino acid
A chain
in order
to find
peptide
mediated
cell
heparin-binding regulating
of
the
attachment
RETROVIRUS-MEDIATED USING <
PROMOTER
BETA-GALACTOSIDASE *1
SH~BA~‘~, Osaka
Shirokansdals have
enables
enough
to
However,
us allow
in
immunocytochemical cell
types
have
inserted
this
type
made
It
of much
nuclear
virus, easier
YOSHIDA,
Sciences,
Osaka
the which
data
A chain
suggest
that
may be crucial
a for
IN
PRIMARY
CULTURE
A REPORTER
University,
3-2
University
Yamadaoka,
of
4 _6 - L
Tokvo,
the
i t
cell
front present
type
in
this
to
the only
IacZ in
the each
Therefore, gene.
the
mild
system, with
identify
markers.
of
IS culture.
overlaps
difficult
system
and
Primary
using often
type-specific in
cell
are
rather
was each
in
staining
assay
cell
cells ws
X-Gal
slonal
Promoter individual
brain
reporter
of makes
against
When
nucleus,
we using which
immunoh~stochemicallv.
OF CEREBELLAR CULTURE
MASAFUMI YANO,
of
laminin
Furthermore.this
These
CELLS AS
in
of
color
identify
BRAIN SIGNAL
activity
beta-ealactosidase
University
domains.We synthesized
activity.
retrovtrus-mediated
promoter
blue
SYNCHRONOUS DIFFERENTIATION SACHIKO
new
iocatlon
to
in the
of the G domain of
the laminin
Science,
differentlatlon
antIbodies a
located
Japan,
a
staining,
with
IN
Medical
108,
the
cytoplasm,
structural
outgrowth.
of
LOCATION
beta-galactosidase,
the
globule
Research, of
detect
normal
since
locates
Protein
developed
to
them is
have been cloned and sequence.
have heparin-binding
SYSTEM
NUCLEAR
Tokyo
recently
One of
outgrowth.
Institute
Mlnato-ku,
We which
for
565.‘The
8 distinct
and neurite
ASSAY
WITH
Ilnstitute
Suita,
to
terminal
and neurite
and Anomalies.
site.
found
carboxyl
laminin. of laminin
deduced from a cDNA clone
attachment.spreading.
site
cell
form at least
out a heparin-binding
A 15 mer peptide.CKDFLSIELVRGRVK.was
in
the A chain
sequence
Psychiatry.Saga Biology
of Health.Bethesda.Maryland.20892.U.S.A.
localized
globule at the end of the long arm. Recently
of Developmental
AND HIROSHI
Tokyo,
7-3-l
SHIMIZU,
Hongo,
Faculty
Bunkyo-ku,
of
Pharmacoeutical
Tokyo
113,
Japan
Improving a culture method, the organotypic culture of cerebellum of a new born rat could be developed its morphology over the entire system. The following two points were improved: 1) two slices explanted face-to-face on the cortical surface on a membrane filter were cultured at air-water interface of the medium. 2) these slices were treated with synchronizers of cell cycle such as Colcemid and Thymidine. Within 24 hrs after the treatment with a cell cycle synchronizer, the glial cells in the external granule layer were differentiated into like Bergmann and at the same time the granule cells lined along these differentiated dia. glia. This temporal evolution of the morphology was quite similar to that in the molecular layer in vivo. In order to develop the morphology over the entire system, it was necessary to once synchronize the cell cycle in the slice. It also suggest that some macroscopic spatio-temporal order would be appeared in the system during development. Therefore we have investigated spatio-temporal pattern of [Calin in differentiating cultured organ.
School.5-l-l
National
Institute
Nabeshima.Saga.849.Japan. of Dental
Four heparin-binding
*Laboratory
Research.National
domains
Institute
had been
It
is
comprised
of 3084 amino acids.which
several
peptides
from the amino acid
A chain
in order
to find
peptide
mediated
cell
heparin-binding regulating
of
the
attachment
RETROVIRUS-MEDIATED USING <
PROMOTER
BETA-GALACTOSIDASE *1
SH~BA~‘~, Osaka
Shirokansdals have
enables
enough
to
However,
us allow
in
immunocytochemical cell
types
have
inserted
this
type
made
It
of much
nuclear
virus, easier
YOSHIDA,
Sciences,
Osaka
the which
data
A chain
suggest
that
may be crucial
a for
IN
PRIMARY
CULTURE
A REPORTER
University,
3-2
University
Yamadaoka,
of
4 _6 - L
Tokvo,
the
i t
cell
front present
type
in
this
to
the only
IacZ in
the each
Therefore, gene.
the
mild
system, with
identify
markers.
of
IS culture.
overlaps
difficult
system
and
Primary
using often
type-specific in
cell
are
rather
was each
in
staining
assay
cell
cells ws
X-Gal
slonal
Promoter individual
brain
reporter
of makes
against
When
nucleus,
we using which
immunoh~stochemicallv.
OF CEREBELLAR CULTURE
MASAFUMI YANO,
of
laminin
Furthermore.this
These
CELLS AS
in
of
color
identify
BRAIN SIGNAL
activity
beta-ealactosidase
University
domains.We synthesized
activity.
retrovtrus-mediated
promoter
blue
SYNCHRONOUS DIFFERENTIATION SACHIKO
new
iocatlon
to
in the
of the G domain of
the laminin
Science,
differentlatlon
antIbodies a
located
Japan,
a
staining,
with
IN
Medical
108,
the
cytoplasm,
structural
outgrowth.
of
LOCATION
beta-galactosidase,
the
globule
Research, of
detect
normal
since
locates
Protein
developed
to
them is
have been cloned and sequence.
have heparin-binding
SYSTEM
NUCLEAR
Tokyo
recently
One of
outgrowth.
Institute
Mlnato-ku,
We which
for
565.‘The
8 distinct
and neurite
ASSAY
WITH
Ilnstitute
Suita,
to
terminal
and neurite
and Anomalies.
site.
found
carboxyl
laminin. of laminin
deduced from a cDNA clone
attachment.spreading.
site
cell
form at least
out a heparin-binding
A 15 mer peptide.CKDFLSIELVRGRVK.was
in
the A chain
sequence
Psychiatry.Saga Biology
of Health.Bethesda.Maryland.20892.U.S.A.
localized
globule at the end of the long arm. Recently
of Developmental
AND HIROSHI
Tokyo,
7-3-l
SHIMIZU,
Hongo,
Faculty
Bunkyo-ku,
of
Pharmacoeutical
Tokyo
113,
Japan
Improving a culture method, the organotypic culture of cerebellum of a new born rat could be developed its morphology over the entire system. The following two points were improved: 1) two slices explanted face-to-face on the cortical surface on a membrane filter were cultured at air-water interface of the medium. 2) these slices were treated with synchronizers of cell cycle such as Colcemid and Thymidine. Within 24 hrs after the treatment with a cell cycle synchronizer, the glial cells in the external granule layer were differentiated into like Bergmann and at the same time the granule cells lined along these differentiated dia. glia. This temporal evolution of the morphology was quite similar to that in the molecular layer in vivo. In order to develop the morphology over the entire system, it was necessary to once synchronize the cell cycle in the slice. It also suggest that some macroscopic spatio-temporal order would be appeared in the system during development. Therefore we have investigated spatio-temporal pattern of [Calin in differentiating cultured organ.