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The expression of estrogen receptor-α mRNA variant in frontal cortex of the mice during the estrus cycle
S140
Abstracts
detergent solubility of PSD molecules. No such increased solubility was observed for GluR2 in the CA3 of GluR1KO. Furthermore, F-/G-actin ratio and spine density were significantly decreased. These findings suggest that GluR2 is critically involved in NMDAR activity as well as formation and maintenance of postsynaptic macromolecular complexes. doi:10.1016/j.neures.2009.09.693
P2-a32 Loss of cerebellar granule cell AMPA receptors contributes to ataxia in stargazer mice Maya Yamazaki 1 , Masahiro Fukaya 2 , Taisuke Miyazaki 2 , Manabu Abe 1 , Rie Natsume 1 , Masahiko Watanabe 2 , Kenji Sakimura 1 1 2
Cell. Neurobiolo., Brain Res. Inst., Niigata University, Niigata, Japan; Anatomy, Hokkaido University Grad. Sch. Med., Sapporo, Japan
The stargazer (stg) mouse is characterized by ataxic gait, head tossing, and absence seizures. Previous studies showed that cell-surface expression and synaptic targeting of AMPA receptors require the membrane protein stargazin (␥-2), which is deficient in the stg mutant mice. To identify which brain region is responsible for the stargazer ataxia, we generated conditional knockout mice using Cre/loxP recombination. In the ␥-2 GC-KO mice, ␥-2 protein was found to disappear from cerebellar granule cell (GC) layer, accompanied by reduced expression of GluR␣ 2 and GluR␣ 4 proteins. Moreover, they showed ataxic gait and poor performance on rota-rod test. In contrast, ␥-2 PC-KO mice, which lacked ␥-2 protein in Purkinje cells selectively, exhibited no ataxia, although GluR␣ 2 and GluR␣ 3 in Purkinje cell dendrite were decreased. These results suggested that stargazer ataxia was attributed to a loss of ␥-2 and AMPA receptors in cerebellar GC. doi:10.1016/j.neures.2009.09.694
P2-a33 Visualization of receptor trafficking in neuronal primary cilia Toru Misawa, Kentaro Yoshimura, Keishi Narita, Sen Takeda Dept. Anat. Cell Biol., Fac. Med. Univ. of Yamanashi, Chuo, Japan Neurons are basic elements in the nervous system, where synapses are the key device in interneuronal communication. Principally, synaptic transmission takes on all-or-none signaling, which is evoked by an action potential. On the other hand, neurons also possess primary cilia that are considered to function mainly as a sensory organelle and modulate the neuronal function in a gradual manner. Previous studies have shown that primary cilia were enriched in numerous receptors and ion channels. The major determinant of sensory modality is receptors such as melanin-concentrating hormone receptor 1, somatostatin receptor 3 and 5-hydroxytryptamine receptor 6. Here, we have confirmed these receptors localized in neuronal primary cilia. Moreover, we have visualized the receptor trafficking by co-transfection of CFP-tagged receptors and YFP-tagged inversin which is known as a molecule specifically targeted to the primary cilia. Our studies indicated that primary cilia play an important role in sensing the extracellular milieu of neurons, and in turn, modulating the neuronal function in meticulous manner.
P2-a34 Immunohistochemical analyses of the expression of Prrt3 in the mouse brain, an orphan metabotropic receptor in the family C Tomomi Yamamoto 1 , Yoshihiro Kubo 1,2 Div Biophys & Neurobiol, Natl Inst Physiol Sci, Okazaki, Japan; JST, Kawaguchi, Japan
2
SORST,
Mouse Prrt3 (Proline rich transmembrane protein) is an orphan metabotropic receptor in the family C, with a large N-terminal extracellular domain and seven transmembrane regions. In addition to the receptor type- long form (Prrt3-Long), a secreted protein type- short form (Prrt3-Short) is translated by slice variation which consists only of N-terminal extracellular domain. As an initial step to approach the function of these proteins, we raised two specific polyclonal antibodies against the N- or C- terminal region, and analyzed their expression patterns in the mouse. Prrt3Short was expressed exclusively in the choroid plexus epithelial cells in the brain ventricles, and was confirmed to be secreted in the cerebrospinal fluid by Western blotting. Prrt3-Long was expressed in the layer beneath the cerebellar Purkinje cells, presumably at the so called Pinceau consists of presynaptic termini of the Basket cells on the Purkinje cells. It was also expressed at the intercalated disc of the cardiac muscle cells and on the wall of the pancreatic ducts. doi:10.1016/j.neures.2009.09.696
1 Lab. Medicinal Pharmacol, Grad. Sch. Pharmaceut. Sci., Osaka University, Osaka, Japan; 2 RIKEN CMIS, Kobe, Japan; 3 Department of Experimental Disease Model, The Osaka-Hamamatsu Joint Research Center for Child Mental Dev., Grad. Sch. Med. Osaka University, Osaka, Japan
Small animal studies are important for elucidation of the pathophysiological mechanism in several diseases, since many gene-manipulated models have been developed in rodents. Recently, PET system has been dramatically improved its performance, and it allows noninvasive imaging in rodents. We previously established the in vivo [18 F]FDG imaging method by PET for mice without anesthesia, and evaluated the brain glucose utilization was high in conscious state compared with isoflurane anesthesia. Here, we have reported dopamine transporter (DAT) imaging using [11 C]PE2I in mouse under unanesthetized condition. The binding potential of [11 C]PE2I in the striatum was significantly high under the anesthetized state, as compared with that under the conscious state. The result strongly indicates that functional state of DAT in vivo is affected by isoflurane anesthesia. doi:10.1016/j.neures.2009.09.697
P2-a36 Vesicular glutamate transporter 3 (VGLUT3)-positive periglomerular cells in the rat olfactory bulb Fumihiro Shutoh, Kae Yamamoto, Kazue Sanada, Kohei Mise, Haruo Nogami, Setsuji Hisano Laboratory of Neuroendocrinology, Kansei, Behavioral and Brainscience, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Japan A neuronal network formed in the olfactory bulb (OB) is an initial processing module of olfactory information. Periglomerular cells (PGCs) interact with glutamatergic projection neurons through synapses within glomeruli. Many PGCs are GABAergic interneurons. Recently, although some fibers in the glomerulus are shown to be positive for VGLUT 3, the origin of the fibers is yet unknown. We identified VGLUT3positive cells in the PGC layer of the rat OB by immunohistochemistry and in situ hybridization. In the PCG layer there were more VGLUT3-positive cells medially than laterally. Some VGLUT3 mRNA-expressing cells were positive for GAD. These results suggest that some PGC neurons are glutamatergic, contributing to excitatory and/or inhibitory neurotransmission in the glomerulus. doi:10.1016/j.neures.2009.09.698
P2-a37 The expression of estrogen receptor-␣ mRNA variant in frontal cortex of the mice during the estrus cycle Kyoko Teramoto, Yutaka Yamamuro Department Anim. Sci., Nihon University, Kanagawa, Japan
doi:10.1016/j.neures.2009.09.695
1
P2-a35 Effect of isoflurane anesthesia on dopamine transporter function; PET study in mice with [11 C]PE2I Toshiyuki Kawasaki 1,2 , Hiroshi Mizuma 2 , Hiroko Nagata 2 , Hisashi Doi 2 , Toshio Matsuda 1,3 , Hirotaka Onoe 2
We have previously reported that the expression of estrogen receptor-␣ (ER-␣) mRNA variant that deleted from exon 1 to 4 was decreased with estrogen levels in the mouse neuroblastoma. However, the self-regulation of estrogen to wild-type ER-␣ and ER- ␣ variant in the brain during the estrus cycle has remained unclear. The aim of present study was to examine the expression of ER-␣ mRNA variant in frontal cortex of the mice during the estrus cycle. A real-time PCR analysis using primer sets designed to amplify from exons 3 to 4, exons 4 to 5, exons 5 to 6, exons 6 to 7, and exons 7 to 8 of the mouse ER-␣ gene revealed the existence of ER-␣ mRNA variant that lost prior to exon 5. Furthermore, ER-␣ mRNA variant expression was significantly lower at proestrus than at estrus and diestrus. The wild-type ER-␣ expression did not change throughout the estrus cycle. These findings suggested that the incidence of ER-␣ mRNA variant interlocks negatively with estrogen levels in frontal cortex of the mice. doi:10.1016/j.neures.2009.09.699
P2-a38 Distinct cellular expression and synaptic localization of NMDA receptor GluN2B and GluN2D subunits in the mouse somatosensory cortex Masahiko Watanabe, Masahiro Fukaya, Miwako Yamasaki Hokkaido University, Japan Of two major NMDA receptor GluN2 subunits expressed in neonates, GluN2B, but not GluN2D, plays an essential role in whisker-related central patterning. Here we compared expression of these GluN2 subunits in the somatosensory cortex. In adult
Abstracts
detergent solubility of PSD molecules. No such increased solubility was observed for GluR2 in the CA3 of GluR1KO. Furthermore, F-/G-actin ratio and spine density were significantly decreased. These findings suggest that GluR2 is critically involved in NMDAR activity as well as formation and maintenance of postsynaptic macromolecular complexes. doi:10.1016/j.neures.2009.09.693
P2-a32 Loss of cerebellar granule cell AMPA receptors contributes to ataxia in stargazer mice Maya Yamazaki 1 , Masahiro Fukaya 2 , Taisuke Miyazaki 2 , Manabu Abe 1 , Rie Natsume 1 , Masahiko Watanabe 2 , Kenji Sakimura 1 1 2
Cell. Neurobiolo., Brain Res. Inst., Niigata University, Niigata, Japan; Anatomy, Hokkaido University Grad. Sch. Med., Sapporo, Japan
The stargazer (stg) mouse is characterized by ataxic gait, head tossing, and absence seizures. Previous studies showed that cell-surface expression and synaptic targeting of AMPA receptors require the membrane protein stargazin (␥-2), which is deficient in the stg mutant mice. To identify which brain region is responsible for the stargazer ataxia, we generated conditional knockout mice using Cre/loxP recombination. In the ␥-2 GC-KO mice, ␥-2 protein was found to disappear from cerebellar granule cell (GC) layer, accompanied by reduced expression of GluR␣ 2 and GluR␣ 4 proteins. Moreover, they showed ataxic gait and poor performance on rota-rod test. In contrast, ␥-2 PC-KO mice, which lacked ␥-2 protein in Purkinje cells selectively, exhibited no ataxia, although GluR␣ 2 and GluR␣ 3 in Purkinje cell dendrite were decreased. These results suggested that stargazer ataxia was attributed to a loss of ␥-2 and AMPA receptors in cerebellar GC. doi:10.1016/j.neures.2009.09.694
P2-a33 Visualization of receptor trafficking in neuronal primary cilia Toru Misawa, Kentaro Yoshimura, Keishi Narita, Sen Takeda Dept. Anat. Cell Biol., Fac. Med. Univ. of Yamanashi, Chuo, Japan Neurons are basic elements in the nervous system, where synapses are the key device in interneuronal communication. Principally, synaptic transmission takes on all-or-none signaling, which is evoked by an action potential. On the other hand, neurons also possess primary cilia that are considered to function mainly as a sensory organelle and modulate the neuronal function in a gradual manner. Previous studies have shown that primary cilia were enriched in numerous receptors and ion channels. The major determinant of sensory modality is receptors such as melanin-concentrating hormone receptor 1, somatostatin receptor 3 and 5-hydroxytryptamine receptor 6. Here, we have confirmed these receptors localized in neuronal primary cilia. Moreover, we have visualized the receptor trafficking by co-transfection of CFP-tagged receptors and YFP-tagged inversin which is known as a molecule specifically targeted to the primary cilia. Our studies indicated that primary cilia play an important role in sensing the extracellular milieu of neurons, and in turn, modulating the neuronal function in meticulous manner.
P2-a34 Immunohistochemical analyses of the expression of Prrt3 in the mouse brain, an orphan metabotropic receptor in the family C Tomomi Yamamoto 1 , Yoshihiro Kubo 1,2 Div Biophys & Neurobiol, Natl Inst Physiol Sci, Okazaki, Japan; JST, Kawaguchi, Japan
2
SORST,
Mouse Prrt3 (Proline rich transmembrane protein) is an orphan metabotropic receptor in the family C, with a large N-terminal extracellular domain and seven transmembrane regions. In addition to the receptor type- long form (Prrt3-Long), a secreted protein type- short form (Prrt3-Short) is translated by slice variation which consists only of N-terminal extracellular domain. As an initial step to approach the function of these proteins, we raised two specific polyclonal antibodies against the N- or C- terminal region, and analyzed their expression patterns in the mouse. Prrt3Short was expressed exclusively in the choroid plexus epithelial cells in the brain ventricles, and was confirmed to be secreted in the cerebrospinal fluid by Western blotting. Prrt3-Long was expressed in the layer beneath the cerebellar Purkinje cells, presumably at the so called Pinceau consists of presynaptic termini of the Basket cells on the Purkinje cells. It was also expressed at the intercalated disc of the cardiac muscle cells and on the wall of the pancreatic ducts. doi:10.1016/j.neures.2009.09.696
1 Lab. Medicinal Pharmacol, Grad. Sch. Pharmaceut. Sci., Osaka University, Osaka, Japan; 2 RIKEN CMIS, Kobe, Japan; 3 Department of Experimental Disease Model, The Osaka-Hamamatsu Joint Research Center for Child Mental Dev., Grad. Sch. Med. Osaka University, Osaka, Japan
Small animal studies are important for elucidation of the pathophysiological mechanism in several diseases, since many gene-manipulated models have been developed in rodents. Recently, PET system has been dramatically improved its performance, and it allows noninvasive imaging in rodents. We previously established the in vivo [18 F]FDG imaging method by PET for mice without anesthesia, and evaluated the brain glucose utilization was high in conscious state compared with isoflurane anesthesia. Here, we have reported dopamine transporter (DAT) imaging using [11 C]PE2I in mouse under unanesthetized condition. The binding potential of [11 C]PE2I in the striatum was significantly high under the anesthetized state, as compared with that under the conscious state. The result strongly indicates that functional state of DAT in vivo is affected by isoflurane anesthesia. doi:10.1016/j.neures.2009.09.697
P2-a36 Vesicular glutamate transporter 3 (VGLUT3)-positive periglomerular cells in the rat olfactory bulb Fumihiro Shutoh, Kae Yamamoto, Kazue Sanada, Kohei Mise, Haruo Nogami, Setsuji Hisano Laboratory of Neuroendocrinology, Kansei, Behavioral and Brainscience, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Japan A neuronal network formed in the olfactory bulb (OB) is an initial processing module of olfactory information. Periglomerular cells (PGCs) interact with glutamatergic projection neurons through synapses within glomeruli. Many PGCs are GABAergic interneurons. Recently, although some fibers in the glomerulus are shown to be positive for VGLUT 3, the origin of the fibers is yet unknown. We identified VGLUT3positive cells in the PGC layer of the rat OB by immunohistochemistry and in situ hybridization. In the PCG layer there were more VGLUT3-positive cells medially than laterally. Some VGLUT3 mRNA-expressing cells were positive for GAD. These results suggest that some PGC neurons are glutamatergic, contributing to excitatory and/or inhibitory neurotransmission in the glomerulus. doi:10.1016/j.neures.2009.09.698
P2-a37 The expression of estrogen receptor-␣ mRNA variant in frontal cortex of the mice during the estrus cycle Kyoko Teramoto, Yutaka Yamamuro Department Anim. Sci., Nihon University, Kanagawa, Japan
doi:10.1016/j.neures.2009.09.695
1
P2-a35 Effect of isoflurane anesthesia on dopamine transporter function; PET study in mice with [11 C]PE2I Toshiyuki Kawasaki 1,2 , Hiroshi Mizuma 2 , Hiroko Nagata 2 , Hisashi Doi 2 , Toshio Matsuda 1,3 , Hirotaka Onoe 2
We have previously reported that the expression of estrogen receptor-␣ (ER-␣) mRNA variant that deleted from exon 1 to 4 was decreased with estrogen levels in the mouse neuroblastoma. However, the self-regulation of estrogen to wild-type ER-␣ and ER- ␣ variant in the brain during the estrus cycle has remained unclear. The aim of present study was to examine the expression of ER-␣ mRNA variant in frontal cortex of the mice during the estrus cycle. A real-time PCR analysis using primer sets designed to amplify from exons 3 to 4, exons 4 to 5, exons 5 to 6, exons 6 to 7, and exons 7 to 8 of the mouse ER-␣ gene revealed the existence of ER-␣ mRNA variant that lost prior to exon 5. Furthermore, ER-␣ mRNA variant expression was significantly lower at proestrus than at estrus and diestrus. The wild-type ER-␣ expression did not change throughout the estrus cycle. These findings suggested that the incidence of ER-␣ mRNA variant interlocks negatively with estrogen levels in frontal cortex of the mice. doi:10.1016/j.neures.2009.09.699
P2-a38 Distinct cellular expression and synaptic localization of NMDA receptor GluN2B and GluN2D subunits in the mouse somatosensory cortex Masahiko Watanabe, Masahiro Fukaya, Miwako Yamasaki Hokkaido University, Japan Of two major NMDA receptor GluN2 subunits expressed in neonates, GluN2B, but not GluN2D, plays an essential role in whisker-related central patterning. Here we compared expression of these GluN2 subunits in the somatosensory cortex. In adult