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A monoclonal antibody that labels motoneurons and sensory ganglion cells in the rat nervous system
S86
708
DISTRIBUTIONOF NITRIC OXIDESYNTHASEIN THE FOREBRAINOF MACACA FUSCATA: K. SATOHl, R. ARAI2, K. IKEMOTOl, M. NARITAl, T. NAGA@, AND K. KITAHAMA4. 1Denartment of Psvchiatrv. Shiea University of Medical Science. Otsu; 2Denar tmne t of Anatomy. Fuiita Health Universitv, Tovoake : 3Dept. of Pedi@rirs. Osaka University Medical School, 4DeDartment of ExDerimental Medicine. Universitv of Claude Bernard. Lyon. France.
The distribution of nitric oxide synthase (NOS)-immunoreactive neurons were studied in the forebrain of the Macaca fuscata by ABC immunohistochemistry using antiserum against neuronal NOS (Ohshima et al., 1992). In the forebrain, numerous NOS immunoreactive neurons were observed in the neocortex, caudate nucleus, putamen, nucleus accumbens, nucleus of the diagonal band, anterior perforated substance, and a few in the amygdaloid complex and hippocampal formation. Whereas most of these neurons were labeled extremely intense for NOS, there were also many moderately intense NOS neurons in the substantia innominata, globus pallidus, anza peduncularis and lateral hypothalamic nucleus. This neuroanatomical evidence that NOS are localized in variety of specific transmitter systems further indicates the neural messenger role of nitric oxide in the central nervous system.
709
REGULATION OF STRESS-INDUCED ACCUMULATION OF hsp27 AND aB CRYSTALLIN IN C6 CELLS. 1. MODULATION BY INHIBITORS OR ACTIVATORS OF ARACHIDONATE CASCADE. HlDFNORl lT0. YUTAKA lN&%lMA. KAORl HASPWWA. OSqMv KDTa KAKATO,K&& I Prefect.. 480 *03. daoan.
Regulation of expression of the small stress proteins, hsp27 and aB crystallin, in C6 glioma cells was analyzed by measuring levels of the two protein with specific immunoassays. Levels of the two proteins increased after exposure of cells to arsenite (100 pM for 1 h) or heat (43OC for 15 min) stress. When cells were exposed to the above stress in the presence of indomethacin (an inhibitor of cyclooxygenase) or nordihydroguaiaretic acid (an inhibitor of lipoxygenase), induction of hsp27 and aB crystallin was markedly enhanced. Coexistence of melittin (an activator of phospholipase As), together with either of the above inhibitor, further enhanced induction of the two proteins. However, exposure of cells to melittin with indomethacin at 37OC did not induce the two proteins. In contrast, quinacnne, which is known as an inhibitor of phospholipase A2 suppressed the induction of the two protein by arsenite. The responses of hsp70 to the inhibitors or activators were similar to those of hsp27, which were estimated by Western Wetting test of the same extracts. These results suggest that the stress-induced responses of hsp27 and aB crystallin (and hsp70), are modulated by arachidonate cascade, and that the induction is enhanced under conditions in which the level of free arachidonic acid is elevated in cells.
710
A MONOCLONAL ANTIBODY THAT LABELS MOTONEURONS AND SENSORY GANGLION CELLS IN THE RAT NERVOUS SYSTEM. TAKASHI MAEHARA, KATSUHIKO ONO, KIMIKO TSUTSUI AND AKIRA TOKUNAGA. The Third Department of Anatomy, Okayama University Medical School. 2-5-l Shikata-Cho, Okayama 700, Japan. A monoclonal antibody (MAb5G6), generated by immunizing Balb/c mice with homogenized bovine retinal tissue, labeled the optic nerve fibers in the bovine and rat retinae. In the adult rat brain, MAb-5G6 immunostained intensely large motoneurons in the motor nuclei of the cranial nerve (nucl. III, IV, V, VI. VII and XII, the dorsal motor nucl. of vagus nerve and nucl. ambiguus) and the spinal motoneurons. The immunoreactivity to the MAb was shown in soma and processes but not in the cell nuclei. MAb5G6 labeled also large round sensory cells, such as the nucl. of the mesencephalic trigeminal tract, the trigeminal and spinal ganglia. Furthermore, fibers of the spinal and mesencephalic trigeminal tracts, and of the posterior funiculus were also immunoreactive to MAb-5G6. The MAb was classified as IgM. Western blot analysis of the spinal cord tissue revealed that MAb-5G6 recognized a polypeptide with mol. wt. of 85kDa. In the prenatal rats, immunoreactivity to the MAb was already found in the spinal ganglion cells by the 13th embryonic day. On the other hand, the spinal motoneurons were not labeled by the MAb up to the 18th embryonic day. The present findings suggest that MAb-5G6 is a useful morphological marker for neurons with direct axonal contact with peripheral organs.
708
DISTRIBUTIONOF NITRIC OXIDESYNTHASEIN THE FOREBRAINOF MACACA FUSCATA: K. SATOHl, R. ARAI2, K. IKEMOTOl, M. NARITAl, T. NAGA@, AND K. KITAHAMA4. 1Denartment of Psvchiatrv. Shiea University of Medical Science. Otsu; 2Denar tmne t of Anatomy. Fuiita Health Universitv, Tovoake : 3Dept. of Pedi@rirs. Osaka University Medical School, 4DeDartment of ExDerimental Medicine. Universitv of Claude Bernard. Lyon. France.
The distribution of nitric oxide synthase (NOS)-immunoreactive neurons were studied in the forebrain of the Macaca fuscata by ABC immunohistochemistry using antiserum against neuronal NOS (Ohshima et al., 1992). In the forebrain, numerous NOS immunoreactive neurons were observed in the neocortex, caudate nucleus, putamen, nucleus accumbens, nucleus of the diagonal band, anterior perforated substance, and a few in the amygdaloid complex and hippocampal formation. Whereas most of these neurons were labeled extremely intense for NOS, there were also many moderately intense NOS neurons in the substantia innominata, globus pallidus, anza peduncularis and lateral hypothalamic nucleus. This neuroanatomical evidence that NOS are localized in variety of specific transmitter systems further indicates the neural messenger role of nitric oxide in the central nervous system.
709
REGULATION OF STRESS-INDUCED ACCUMULATION OF hsp27 AND aB CRYSTALLIN IN C6 CELLS. 1. MODULATION BY INHIBITORS OR ACTIVATORS OF ARACHIDONATE CASCADE. HlDFNORl lT0. YUTAKA lN&%lMA. KAORl HASPWWA. OSqMv KDTa KAKATO,K&& I Prefect.. 480 *03. daoan.
Regulation of expression of the small stress proteins, hsp27 and aB crystallin, in C6 glioma cells was analyzed by measuring levels of the two protein with specific immunoassays. Levels of the two proteins increased after exposure of cells to arsenite (100 pM for 1 h) or heat (43OC for 15 min) stress. When cells were exposed to the above stress in the presence of indomethacin (an inhibitor of cyclooxygenase) or nordihydroguaiaretic acid (an inhibitor of lipoxygenase), induction of hsp27 and aB crystallin was markedly enhanced. Coexistence of melittin (an activator of phospholipase As), together with either of the above inhibitor, further enhanced induction of the two proteins. However, exposure of cells to melittin with indomethacin at 37OC did not induce the two proteins. In contrast, quinacnne, which is known as an inhibitor of phospholipase A2 suppressed the induction of the two protein by arsenite. The responses of hsp70 to the inhibitors or activators were similar to those of hsp27, which were estimated by Western Wetting test of the same extracts. These results suggest that the stress-induced responses of hsp27 and aB crystallin (and hsp70), are modulated by arachidonate cascade, and that the induction is enhanced under conditions in which the level of free arachidonic acid is elevated in cells.
710
A MONOCLONAL ANTIBODY THAT LABELS MOTONEURONS AND SENSORY GANGLION CELLS IN THE RAT NERVOUS SYSTEM. TAKASHI MAEHARA, KATSUHIKO ONO, KIMIKO TSUTSUI AND AKIRA TOKUNAGA. The Third Department of Anatomy, Okayama University Medical School. 2-5-l Shikata-Cho, Okayama 700, Japan. A monoclonal antibody (MAb5G6), generated by immunizing Balb/c mice with homogenized bovine retinal tissue, labeled the optic nerve fibers in the bovine and rat retinae. In the adult rat brain, MAb-5G6 immunostained intensely large motoneurons in the motor nuclei of the cranial nerve (nucl. III, IV, V, VI. VII and XII, the dorsal motor nucl. of vagus nerve and nucl. ambiguus) and the spinal motoneurons. The immunoreactivity to the MAb was shown in soma and processes but not in the cell nuclei. MAb5G6 labeled also large round sensory cells, such as the nucl. of the mesencephalic trigeminal tract, the trigeminal and spinal ganglia. Furthermore, fibers of the spinal and mesencephalic trigeminal tracts, and of the posterior funiculus were also immunoreactive to MAb-5G6. The MAb was classified as IgM. Western blot analysis of the spinal cord tissue revealed that MAb-5G6 recognized a polypeptide with mol. wt. of 85kDa. In the prenatal rats, immunoreactivity to the MAb was already found in the spinal ganglion cells by the 13th embryonic day. On the other hand, the spinal motoneurons were not labeled by the MAb up to the 18th embryonic day. The present findings suggest that MAb-5G6 is a useful morphological marker for neurons with direct axonal contact with peripheral organs.