Single particle tracking analysis of endosome movement in glial cells

s131

CNS NEUROBLASTS

207

CONTAINING

MIGRATE IN THREE-DIMENSIONAL

MATRIX

LAMININ.

ISA0 NAGATA’, HIROAKI ASOU’, H. AND JUNK0

KIMURA-KURODA’

‘Dept. of Cell Biology, Tokyo Metropolitan Institute for Neuroscience, ‘Dept. of Nemo-Cell

GELS WITH EXTRACELLULAR

2-6 Musashidai,

Biology, Tokyo Metropolitan Institute for Gerontology,

We have found that three-dimensional

Fuchu-shi,

Sakaecho,

Tokyo 183-8526,

Itabashi-ku,

Tokyo 173-0015

(3-D) fibrin and collagen gels containing laminin (with entactin. LM) promote the

migration of cerebellar granule cells”. To determine whether or not other CNS neurons migrate in such gels, smaII tissues were dissected from mouse cerebral cortices (E12), rat hippocampus embedded in gel matrices formed of plasminogen-free

(E18) or mouse cerebellar nuclei with white matter (P4),

fibrin (FIB) and collagen-type I (COL) with (FIB-LM, COL-LM) or

without LM, cultured in a serum-free synthetic medium and observed by time-lapse video microscopy.

Single neuroblasts

migrated out from these explants and moved freely in both FIB-LM and COL-L.M, but not FIB and COL alone. The migratory dynamics of neuroblasts in FIB-LM was as usual but that in COL-LM was different. Neuroblasts in COL-LM had unique cytoplasma segmented with some constrictions,

and they appeared to move Iike toothpaste squeezed

in a tube. These results

suggest that LM is essential for the migration of CNS neuroblasts in 3-D gels and may serve to elucidate the migratory 1) Kimura-Kuroda, J. et al. in Neural Development (Uyemura, K. et al. eds.) Springer-Verlag Tokyo (in press)

mechanism of neuroblasts in vivo.

208

SINGLE PARTICLE TRACKING

TOMOMITSU TETSUYA

ICHIKAWAI,

URASAKH,

MAKOTO

SUGURU

YAMADAZ,

ANALYSIS

TETSUYA

OF ENDOSOME

KIMOTOl,

MOVEMENT

NOR10 TAKATAI,

IN GLIAL CELLS

KAZUHIRO

SUZUKIl,

KAWATOl

1Dep. of Biophysics and Life Sciences, Graduate School of Arts and Sciences, Univ. of Tokyo at Komaba, Tokyo 153-8902,2Dep. of Physics, Kyorin Univ. School of Medicine, Mitaka, Tokyo 181-8611

Meguro-ku,

Neurosteroids may act as modulater of glutamate receptor function, thereby contributing to learning and memory depending on emotion. In glial cells, neurosteroid is synthesized from cholesterol which was transported with low density lipoprotein LDL via endosome movement. LDL labeled with fluorescent probe, DiI, was loaded into glial cells. The traffic of endosomes was observed with video fluorescence microscopy and analyzed by single particle tracking technique. After lhour from loading of DiI-LDL, fluorescent endosomes were directionally moving forward (1 II m/set) and backward. There were also some immobile endsomes. When we trace endosome movement over the long time range, endosomes took 3-6 hours to be concentrated around the circumference of nuclei of glial cells. This very slow transport is due to the fact that endosomes were not only unidirectionally moving forward but also moving backward and also being immobile.

MODULATORY EFFECT OF PROSTAGLANDIN Ez ON AXONAL TRANSPORT IN CULTURED MOUSE DORSAL ROOT GANGLION CELLS. INVOLVEMENT OF CYCLIC AMP/PROTEIN KINASE 209 A CASCADE HIROMI HIRUMA’. TAKASHI KATAKURA’, SANAE NISHIDA’, TATSUMI KUSAKABEZ. HIROSHI MARUYAMA3. TOSHIFUMI TAKENAKA1, SUM10 HOKA3, TADASHI KAWAKAMI’ iDept of Physiology, Kitasato Univ Sch of Medicine, Sagamihara 228-8555. ZDept of Anatomy, Yokohama City Univ Sch of Medicine, 3Dept. of Anesthesiology, Kitasato Univ Sch of Medicine, lKokushikan Univ. Sch. of Physical Education The effects were

of prostaglandin

examined

transported

with video-enhanced

particles

microscopy.

permeable

of PGEz on the anterograde H-89 (I PM),

the facilitatory

diminished

These

suggest

in cultured

results DRGs

transport

and retrograde

(IO PM), axonal

of both

that PGE2 facilitates

mouse

of I uM PGEz

directions

and folskolin

effects

of cultured

Application

and retrograde

cPIMP,

A inhibitor

cascade

on axonal

in both anterograde

0 5 mM), a membrane the effect

Ez (PGE2)

Both

the axonal

increased

ofadenylate

In the presence

PGEz and dbcAMP transport

root

of dibutyryl

an activator

transport

dorsal

ganglion

the number c.4MP

cells of the

(dbcAMP,

cyclase,

mimicked

of the protein

on axonal

transport

via c.4MP/

protein

kinase

were kinase

A