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Monoclonal antibody 3C8 recognizes retinal ganglion cells of mammals
S208 MONOCLONAL ANTIBODY 3C8 RECOGNIZES RETINAL GANGLION CELLS OF 1736 MAMMALS. TAKETOSHI WAKABAYASHI, JUN KOSAKA AND YUTAKA FUKUDA, 2nd. Dept. of Physiol., Osaka Univ. Med. Sch. Yamadaoka 2-2 Suita, Osaka, 565, Japan. Flatmount preparations have been frequently used for the analysis of distribution and the morphological identification of retinal ganglion cells (RGCs) . Retrograde labelings were used for the detection of RGCs. The drawbacks of retrograde labeling methods are leakage of dyes from labeled cells, erroneous labeling of other cells and incomplete labeling of RGCs. To overcome these problems, we tried to produce monoclonal antibodies with specifically recognize RGCs on the flatmount preparations. One of these antibodies, 3C8, appeared to stain somas of cats. We verified that in flatmount preparations of rat retina, RGCs of rats and labeled cells overlapped with cells stained with 3C8. Even over 90% of retrogradelly in axotomized rat retina, survived RGCs were labeled with 3C8, without erroneous labeling of glial cells. We conclude that monoclonal antibody 3C8 is useful to label RGCs in axotomized retina as well as intact retina.
CREB Phosphorylation in Rat Retinal Cells by Light and Calcium Influx JUNK0 IMAKI, Dept. of Anatomy, Nippon Medical School, 1-1-5, Sendagi, Bunkyo-ku, Tokyo, 113, Japan 1737
The expression of c-fos mRNA and the phosphorylation of CAMP responsive element binding protein (CREB) were examined in the rat retina by in situ hybridization histochemistry and imm.unocytochemistry. c-fos mRNA was expressed in the outer half of the inner nuclear layer (INL) and the ganglion cell layer (GCL) after 30 min of sustained light. After 30 min of flashing light, cfos expression was stronger in the inner border of the INL. Following administration of L-type calcium channel activator Bay K 8644, c-fos expression was detected in the outer nuclear layer was detected in the (ONL), at the inner border of the INL and the GCL. CREB immunoreactivity nuclei in the INL and the GCL. Phosphorylated CREB immunoreactive nuclei had similar distributior, as c-fos mRNA following steady and flashing light, as well as that following Bay K 8644. These results suggest that calcium influx and phosphorylation of CREB may be involved in the c-fos expression in retinal cells in vivo.
1738
APPEARANCE AND MATURATION OF RESPONSES OF EXCITATORY AND INHIBITORY AMINO ACIDS DURING CHIKAFUMI CHIBA AND TAKEHIKO SAITO, Inst, of Biol.
SOLITARY SPIKING CELLS TO NEWT RETINAL REGENERATION.
Sci., Universitv of Tsukuba,
Tsukuba, Ibaraki 305, Japan. Responsiveness to excitatory and inhibitory amino acids in solitary spiking cells dissociated from different stages of regenerating newt retinas were studied with whole-cell patch-clamp methods in comparison with that in the normal retina. In the normal retina, we identified AMPA/kainate and NMDA receptors as excitatory amino acid receptors, and GABAA and glycine receptors as inhibitory amino acid receptors. All spiking cells examined responded to kainate, GABA and glycine, about 70% of them responded to AMPA, and about 50% to NMDA. During retinal regeneration, appearance rate of kainate, GABA and glycine responses monotonically increased to that in the normal retina. While appearance rate of AMPA and NMDA responses temporarily increased to more than the normal level before the regenerating retina has segregated into two synaptic layers, and decreased to the normal level after the segregation of the synaptic layers.
CREB Phosphorylation in Rat Retinal Cells by Light and Calcium Influx JUNK0 IMAKI, Dept. of Anatomy, Nippon Medical School, 1-1-5, Sendagi, Bunkyo-ku, Tokyo, 113, Japan 1737
The expression of c-fos mRNA and the phosphorylation of CAMP responsive element binding protein (CREB) were examined in the rat retina by in situ hybridization histochemistry and imm.unocytochemistry. c-fos mRNA was expressed in the outer half of the inner nuclear layer (INL) and the ganglion cell layer (GCL) after 30 min of sustained light. After 30 min of flashing light, cfos expression was stronger in the inner border of the INL. Following administration of L-type calcium channel activator Bay K 8644, c-fos expression was detected in the outer nuclear layer was detected in the (ONL), at the inner border of the INL and the GCL. CREB immunoreactivity nuclei in the INL and the GCL. Phosphorylated CREB immunoreactive nuclei had similar distributior, as c-fos mRNA following steady and flashing light, as well as that following Bay K 8644. These results suggest that calcium influx and phosphorylation of CREB may be involved in the c-fos expression in retinal cells in vivo.
1738
APPEARANCE AND MATURATION OF RESPONSES OF EXCITATORY AND INHIBITORY AMINO ACIDS DURING CHIKAFUMI CHIBA AND TAKEHIKO SAITO, Inst, of Biol.
SOLITARY SPIKING CELLS TO NEWT RETINAL REGENERATION.
Sci., Universitv of Tsukuba,
Tsukuba, Ibaraki 305, Japan. Responsiveness to excitatory and inhibitory amino acids in solitary spiking cells dissociated from different stages of regenerating newt retinas were studied with whole-cell patch-clamp methods in comparison with that in the normal retina. In the normal retina, we identified AMPA/kainate and NMDA receptors as excitatory amino acid receptors, and GABAA and glycine receptors as inhibitory amino acid receptors. All spiking cells examined responded to kainate, GABA and glycine, about 70% of them responded to AMPA, and about 50% to NMDA. During retinal regeneration, appearance rate of kainate, GABA and glycine responses monotonically increased to that in the normal retina. While appearance rate of AMPA and NMDA responses temporarily increased to more than the normal level before the regenerating retina has segregated into two synaptic layers, and decreased to the normal level after the segregation of the synaptic layers.