- Email: [email protected]
1020 Production and characterization of a monoclonal antibody against the green fluorescent protein (GFP)
s122
1018
ORIGIN AND MIGRATION OF THE OPTIC NERVE OLIGODENDROCYTE IN THE CHICK EMBRYO Dept. of Anatomy (2nd Division), Shimane Med. Univ., Izumo 693. Japan’, Dept. of Psychiatry, Shimane Med. Univ., Izumo 693, Japan” KATSUHIKO
ONO’, TOSHIKO
TSUMORI’,
TOSHIRO
KISHI”,
YUKIHIKO YASUI’ The origin and migration of the optic nerve oligodendrocyte (OL) were studied in the chick embryo with monoclonal antibodies 01 and 04, which label the OL and its precursor cell, respectively. 04+ cells were initially observed in the midline ventricular layer (VL) just dorsal to the optic chasm at 5th day of incubation (E5). Then, 04+ cells spread to the chiasmal end of the optic nerve, and arrived at the retinal end by E9. At ElO, 04-t cells appeared in the retina. Cultured explants of the chiasmal region from E5 or E6 generated Ol+ OL. Two days after intraventricular injection of DiI to E6 embryos, the VL of the third ventricle was intensely labeled with the dye, and cells labeled with DiI were observed in the optic nerve, some of which were 04+. The present results suggest that the OL in the optic nerve arises from the VL of the thrid ventricle, probably from the bottom of the midline region, and that they migrate in the optic nerve towards the retina.
1019
EFFECTS OF NEONATAL STRESS ON POSTNATAL DEVELOPMENT OF THE LIMBIC SYSTEM AND LOCOMOTOR FUNCTION Dept. Physiol., Fat. Med., Toyama Med. & Pharmaceu. Univ., Sugitani 2630, Toyama 930-01, Japan Teruko Uwano, Hisao Nishijo, Kyoko Tanebe, Ryoi Tamura, Taketoshi Ono Repeated cold stress was loaded to pregnant rats from 10 to 20 day gestation age, and their offspring were used. At postnatal day (PND) 8, the mean body weight, and the mean wet weights of the brain, thymus,,spleen, and kidney were smaller in the stress-loaded pups than those of the non-stressed control pups. The mean breadth of area of the anterior cingulate cortex was also smaller in the stress-loaded pups. From PND 6 to 14, swimming in warm water and duration for turning of the body axis from below upward on a inclined plane after pups were put upside down were also analyzed. Control pups at PND 6 swam smoothly with nose-up, and regular and sequential movements of extremities. The stress-loaded pups bent the body axis to right or left, and could not swim straightway. The mean duration for body turning were larger in stress-loaded pups. These results suggest that neonatal stress could widely affect postnatal functional and anatomical development.
1020
PRODUCTION AND CHARACTERIZATION OF A MONOCLONAL AGAINST THE GREEN FLUORESCENT PROTEIN (GFP)
Dept. Physiol. Tokyo Women’s Medical College, 8-l Kawada-cho, Shinjuku-ku,
ANTIBODY
Tokyo 162, Japan
Keiko Gengyo-Ando, Shohei Mitani GFP is a unique ln vivo reporter for analyzing gene expression in many species. We have generated monoclonal antibodies against GFP using a GST fusion protein containing the full-length of GFP coding region and a synthetic peptide corresponding to internal 208Ser - 217His residues of GFP as immunogens. By immunoblot analysis, they specifically recognized the fusion proteins expressed in E. coli, and one of the antibodies, 65B12, was also available for immunohistochemistry, since it recognized the fluorescing cells observed in living animals transgenic to an uric-86-gfp reporter construct. The indirect immunostaining using a rodamine-conjugated secondary antibody showed greater brightness than a GFP variant (GFP-TT, a gift from Jin and Horvitz) observed in living cells. Thus, this antibody against GFP will be helpful for the identification of the GFP expressing cells even if they make relatively less brightness. This work was done in collaboration with Aki Yonezumi.
1018
ORIGIN AND MIGRATION OF THE OPTIC NERVE OLIGODENDROCYTE IN THE CHICK EMBRYO Dept. of Anatomy (2nd Division), Shimane Med. Univ., Izumo 693. Japan’, Dept. of Psychiatry, Shimane Med. Univ., Izumo 693, Japan” KATSUHIKO
ONO’, TOSHIKO
TSUMORI’,
TOSHIRO
KISHI”,
YUKIHIKO YASUI’ The origin and migration of the optic nerve oligodendrocyte (OL) were studied in the chick embryo with monoclonal antibodies 01 and 04, which label the OL and its precursor cell, respectively. 04+ cells were initially observed in the midline ventricular layer (VL) just dorsal to the optic chasm at 5th day of incubation (E5). Then, 04+ cells spread to the chiasmal end of the optic nerve, and arrived at the retinal end by E9. At ElO, 04-t cells appeared in the retina. Cultured explants of the chiasmal region from E5 or E6 generated Ol+ OL. Two days after intraventricular injection of DiI to E6 embryos, the VL of the third ventricle was intensely labeled with the dye, and cells labeled with DiI were observed in the optic nerve, some of which were 04+. The present results suggest that the OL in the optic nerve arises from the VL of the thrid ventricle, probably from the bottom of the midline region, and that they migrate in the optic nerve towards the retina.
1019
EFFECTS OF NEONATAL STRESS ON POSTNATAL DEVELOPMENT OF THE LIMBIC SYSTEM AND LOCOMOTOR FUNCTION Dept. Physiol., Fat. Med., Toyama Med. & Pharmaceu. Univ., Sugitani 2630, Toyama 930-01, Japan Teruko Uwano, Hisao Nishijo, Kyoko Tanebe, Ryoi Tamura, Taketoshi Ono Repeated cold stress was loaded to pregnant rats from 10 to 20 day gestation age, and their offspring were used. At postnatal day (PND) 8, the mean body weight, and the mean wet weights of the brain, thymus,,spleen, and kidney were smaller in the stress-loaded pups than those of the non-stressed control pups. The mean breadth of area of the anterior cingulate cortex was also smaller in the stress-loaded pups. From PND 6 to 14, swimming in warm water and duration for turning of the body axis from below upward on a inclined plane after pups were put upside down were also analyzed. Control pups at PND 6 swam smoothly with nose-up, and regular and sequential movements of extremities. The stress-loaded pups bent the body axis to right or left, and could not swim straightway. The mean duration for body turning were larger in stress-loaded pups. These results suggest that neonatal stress could widely affect postnatal functional and anatomical development.
1020
PRODUCTION AND CHARACTERIZATION OF A MONOCLONAL AGAINST THE GREEN FLUORESCENT PROTEIN (GFP)
Dept. Physiol. Tokyo Women’s Medical College, 8-l Kawada-cho, Shinjuku-ku,
ANTIBODY
Tokyo 162, Japan
Keiko Gengyo-Ando, Shohei Mitani GFP is a unique ln vivo reporter for analyzing gene expression in many species. We have generated monoclonal antibodies against GFP using a GST fusion protein containing the full-length of GFP coding region and a synthetic peptide corresponding to internal 208Ser - 217His residues of GFP as immunogens. By immunoblot analysis, they specifically recognized the fusion proteins expressed in E. coli, and one of the antibodies, 65B12, was also available for immunohistochemistry, since it recognized the fluorescing cells observed in living animals transgenic to an uric-86-gfp reporter construct. The indirect immunostaining using a rodamine-conjugated secondary antibody showed greater brightness than a GFP variant (GFP-TT, a gift from Jin and Horvitz) observed in living cells. Thus, this antibody against GFP will be helpful for the identification of the GFP expressing cells even if they make relatively less brightness. This work was done in collaboration with Aki Yonezumi.