TY - JOUR AB - Microglia have emerged as a critical component of neurodegenerative diseases. Genetic manipulation of microglia can elucidate their functional impact in disease. In neuroscience, recombinant viruses such as lentiviruses and adeno-associated viruses (AAVs) have been successfully used to target various cell types in the brain, although effective transduction of microglia is rare. In this review, we provide a short background of lentiviruses and AAVs, and strategies for designing recombinant viral vectors. Then, we will summarize recent literature on successful microglial transductions in vitro and in vivo, and discuss the current challenges. Finally, we provide guidelines for reporting the efficiency and specificity of viral targeting in microglia, which will enable the microglial research community to assess and improve methodologies for future studies. AU - Maes, Margaret E AU - Colombo, Gloria AU - Schulz, Rouven AU - Siegert, Sandra ID - 6521 JF - Neuroscience Letters SN - 0304-3940 TI - Targeting microglia with lentivirus and AAV: Recent advances and remaining challenges VL - 707 ER - TY - JOUR AB - We examined immunoreactivities for γ-aminobutyric acidB-receptor (GABABR) subtypes, GABABR1 and GABABR2, in the mesencephalic trigeminal nucleus neurons (MTN neurons) of the rat. Immunoreactivity for GABABR1 was prominent in cell bodies of MTN, whereas that for GABABR2 was very weak, if existed. For electron microscopy, the immunogold-silver method for GABABR1 was combined with the immunoperoxidase method for glutamic acid decarboxylase (GAD: the synthetic enzyme of GABA). Immunogold-silver particles indicating GABABR1 immunoreactivity were distributed widely in the cytoplasm of the cell bodies postsynaptic to GAD-immunoreactive axon terminals, but were rarely associated with synaptic membrane specialization or extrasynaptic sites of plasma membrane. It has been indicated that GABABR1 may not be transported to plasma membrane when no GABABR2 exists. Thus, it was presumed that GABABR1 in the cell body of the rat MTN neurons might not be involved in the synaptic transmission. AU - Li, Jin AU - Shigemoto, Ryuichi AU - Kulik, Ákos AU - Chen, Peng AU - Nomura, Sakashi AU - Kaneko, Takeshi AU - Mizuno, Noboru ID - 2612 IS - 1-2 JF - Neuroscience Letters SN - 0304-3940 TI - Immunocytochemical localization of GABAB receptors in mesencephalic trigeminal nucleus neurons in the rat VL - 315 ER - TY - JOUR AB - Substance P receptor (i.e. NK1)-like immunoreactive (SPR-LI) neurons were observed in the newborn and adult human spinal cord. Substance P receptor-like immunoreactive neuronal cell bodies were seen most frequently in lamina I, and were scattered throughout the remaining laminae of the dorsal horn and the area around the central canal. Some neurons in the intermediolateral nucleus also showed weak immunoreactivity. The pattern of distribution of SPR-LI neurons in the adult spinal cord was essentially the same as that in the newborn spinal cord. However, SPR-LI neurons cell bodies were seen much more frequently in the newborn than in the adult dorsal horn, especially in lamina II. AU - Ding, Yu AU - Zheng, Heng AU - Wang, Dian AU - Xu, Jun AU - Gong, Liang AU - Lü, Yan AU - Qin, Bing AU - Shi, Juan AU - Li, Hua AU - Li, Ji AU - Shigemoto, Ryuichi AU - Kaneko, Takeshi AU - Mizuno, Noboru ID - 2594 IS - 2 JF - Neuroscience Letters SN - 0304-3940 TI - The distribution of substance P receptor (NK1)-like immunoreactive neurons in the newborn and adult human spinal cord VL - 266 ER - TY - JOUR AB - The occurrence and distribution of the preferred receptor for the neuropeptide, substance P (SP), the neurokinin-1 receptor (NK1R) was investigated in the vascular supply of the rat sciatic nerve. Messenger RNA for NK1R was demonstrated by RT-PCR in the epineurial layer where the majority of small arteries and arterioles feeding the endoneurial vasculature are located. Immunoreactivity to NK1R-protein was localized on the smooth muscle cells of these arterial vessels by means of immunofluorescence using a polyclonal NK1R antiserum. This muscular localization of NK1R explains the previously reported [Zochodne, D.W. and Ho, L.T., J. Physiol. 444 (1991) 615- 630] moderate vasoconstrictor rather than vasodilator effects of SP in this vascular bed. AU - Kummer, Wolfgang AU - Shigemoto, Ryuichi AU - Haberberger, Rainer ID - 2591 IS - 2 JF - Neuroscience Letters SN - 0304-3940 TI - Smooth muscle cells are the site of neurokinin-1 receptor localization in the arterial supply of the rat sciatic nerve VL - 259 ER - TY - JOUR AB - It was examined electron microscopically in the rat if a metabotropic glutamate receptor, mGluR7, might be localized in axon terminals of nociceptive, primary afferent fibers in laminae I and II of the spinal dorsal horn. Nociceptive nature of axon terminals showing mGluR7-like immunoreactivity (mGluR7-LI) was indicated by binding to the isolectin I-B4 from Griffonia simplicifolia (I-B4), or by substance P-like immunoreactivity (SP-LI). Axon terminals labeled with immunogold particles indicating mGluR7-LI were usually filled with round synaptic vesicles and were in asymmetric synaptic contact with dendritic or somatic profiles; occasionally they contained pleomorphic vesicles and were in symmetric synaptic contact with somatic profiles in lamina II. The double-labeling studies revealed that most of axon terminals with I-B4 labeling as well as a small population of axon terminals with SP-LI, showed mGluR7-LI. About one-third or much smaller population of axon terminals with mGluR7-LI in laminae I and II were labeled, respectively, with I-B4 or SP-LI; these were in asymmetric synaptic contact with dendritic profiles. AU - Li, He AU - Ohishi, Hitoshi AU - Kinoshita, Ayae AU - Shigemoto, Ryuichi AU - Nomura, Sakashi AU - Mizuno, Noboru ID - 2575 IS - 3 JF - Neuroscience Letters SN - 0304-3940 TI - Localization of a metabotropic glutamate receptor, mGluR7, in axon terminals of presumed nociceptive, primary afferent fibers in the superficial layers of the spinal dorsal horn: An electron microscope study in the rat VL - 223 ER - TY - JOUR AB - The present study indicated presynaptic localization of a metabotropic glutamate receptor, mGluR8, in projection neurons of the main olfactory bulb of rat. An antibody was produced by using a peptide corresponding to C-terminal 23 amino acids of mouse mGluR8. It was confirmed that the C-terminal 23 amino acids of rat mGluR8 were the same as those of mouse mGluR8 except for one, and that the antibody specifically recognized mGluR8 in the rat rhinencephalon. In layer Ia of the piriform cortex (a target area of projection fibers from the main olfactory bulb), mGluR8-like immunoreactivity (mGluR8-LI) was reduced after transection of the lateral olfactory tract, and mGluR8-LI was observed in axon terminals which were filled with round synaptic vesicles and made asymmetric synapses with dendritic spines. AU - Kinoshita, Ayae AU - Ohishi, Hitoshi AU - Neki, Akio AU - Nomura, Sakashi AU - Shigemoto, Ryuichi AU - Takada, Masahiko AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2566 IS - 1 JF - Neuroscience Letters SN - 0304-3940 TI - Presynaptic localization of a metabotropic glutamate receptor, mGluR8, in the rhinencephalic areas: A light and electron microscope study in the rat VL - 207 ER - TY - JOUR AB - Immunoreactivity for the metabotropic glutamate receptor 7 (mGluR7) and that for phosphate-activated glutaminase (PAG) were examined in the trigeminal (TG), dorsal root (DRG), nodose (NG), superior cervical, celiac, and pelvic ganglia of the rat. Virtually all neuronal cell bodies showed mGluR7-like immunoreactivity (mGluR7-LI) in these ganglia. On the other hand, PAG-like immunoreactivity (PAG) was seen in almost all neuronal cell bodies in the TG, DRG and NG, but not in the other ganglia. Co-existence of mGluR7- and PAG-LI in the TG, DRG and NG was confirmed by a double-immunofluorescence immunohistochemical method. The results indicate that virtually all sensory ganglion neurons are glutamatergic and equipped with mGluR7. AU - Li, Jin AU - Ohishi, Hitoshi AU - Kaneko, Takeshi AU - Shigemoto, Ryuichi AU - Neki, Akio AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2565 IS - 1-2 JF - Neuroscience Letters SN - 0304-3940 TI - Immunohistochemical localization of a metabotropic glutamate receptor, mGluR7, in ganglion neurons of the rat; with special reference to the presence in glutamatergic ganglion neurons VL - 204 ER - TY - JOUR AB - A monoclonal antibody against a metabotropic glutamate receptor, mGluR2, was produced by using a glutathione S-transferase (GST) fusion protein containing an N-terminal sequence of rat mGluR2. Intense mGluR2-like immunoreactivity (mGluR2-LI) was seen mainly in neuropil of the cerebral cortical regions, hippocampus, olfactory bulb, some diencephalic nuclei, dorsal cochlear nucleus and cerebellar cortex. In the cerebellar cortex, mGluR2-LI was seen only in Golgi cells. In Ammon's hem, mGluR2-LI was marked in the stratum lucidum of CA3 and the stratum lacunosum-moleculare of CA1-CA3, but not detected in the stratum pyramidale. The results indicate that mGluR2 is located not only presynaptically but also postsynaptically. AU - Neki, Akio AU - Ohishi, Hitoshi AU - Kaneko, Takeshi AU - Shigemoto, Ryuichi AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2562 IS - 3 JF - Neuroscience Letters SN - 0304-3940 TI - Pre- and postsynaptic localization of a metabotropic glutamate receptor, mGluR2, in the rat brain: An immunohistochemical study with a monoclonal antibody VL - 202 ER - TY - JOUR AB - Localization of a metabotropic glutamate receptor, mGluR4a, was immunohistochemically examined in the rat cerebellum with an antibody, which was produced by using a synthetic peptide corresponding to a C-terminal sequence of rat mGluR4a. Marked mGluR4a-like immunoreactivity (mGluRLta-LI) was seen in neuropil of the molecular layer of the cerebellar cortex. Electron microscopically, mGluR4a-LI was observed in many axon terminals in the molecular layer. These axon terminals showing mGluR4a-LI were filled with round synaptic vesicles and were in asymmetric synaptic contacts most frequently with dendritic spines. The results indicate that mGluR4a are located presynaptically in the parallel fibers arising from the granule cells in the cerebellar cortex. AU - Kinoshita, Ayae AU - Ohishi, Hitoshi AU - Nomura, Sakashi AU - Shigemoto, Ryuichi AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2568 IS - 3 JF - Neuroscience Letters SN - 0304-3940 TI - Presynaptic localization of a metabotropic glutamate receptor, mGluR4a, in the cerebellar cortex: A light and electron microscope study in the rat VL - 207 ER - TY - JOUR AB - Morphological substrates for interactions between γ-aminobutyric acid (GABA) and substance P upon neurons expressing substance Preceptor (SPR) in the nucleus of the solitary tract (NST) were investigated by immunocytochemical electron microscopy. In the NST of the rat, many GABA-like immunoreactive axon terminals were in symmetric synaptic contacts with dendritic profiles; they were observed on nearly a half of the SPR-like immunoreactive dendritic profiles in the medial part of the caudal half of the NST. AU - Jia, Hong AU - Wang, Bai AU - Rao, Zhi AU - Shi, Ji AU - Shigemoto, Ryuichi AU - Kaneko, Takeshi AU - Mizuno, Noboru ID - 2569 IS - 1 JF - Neuroscience Letters SN - 0304-3940 TI - GABAergic synapses upon neurons expressing substance P receptors in the nucleus of the solitary tract: An immunocytochemical electron microscope study in the rat VL - 210 ER - TY - JOUR AB - An antibody which recognizes specifically a metabotropic glutamate receptor, mGluR7, was produced by using a trpE fusion protein containing a C-terminal sequence of rat mGluR7. Neuropil in laminae I and II of the dorsal horn of the rat, as well as many neuronal cell bodies in the dorsal root ganglion, showed mGluR7-like immunoreactivity; the immunoreactivity in neuropil was seen in axon terminals, which were filled with round synaptic vesicles and constituted axodendritic and axosomatic asymmetric synapses. The mGluR7-like immunoreactivity in laminae I and II in the dorsal horn was reduced after dorsal rhizotomy. The results indicate that some axon terminals of the primary afferent fibers to laminae I and II of the dorsal horn are provided with mGluR7. AU - Ohishi, Hitoshi AU - Nomura, Sakashi AU - Ding, Yu AU - Shigemoto, Ryuichi AU - Wada, Eiki AU - Kinoshita, Ayae AU - Li, Jin AU - Neki, Akio AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2561 IS - 1-2 JF - Neuroscience Letters SN - 0304-3940 TI - Presynaptic localization of a metabotropic glutamate receptor, mGluR7, in the primary afferent neurons: An immunohistochemical study in the rat VL - 202 ER - TY - JOUR AB - Chemical irritation of the urinary bladder with formalin in the rat induced c-fos protein-like immunoreactivity in more than 80% of substance P receptor-like immunoreactive (SPR-LI) neurons of the dorsal commissural nucleus, sacral parasympathetic nucleus and lamina I in the 6th lumbar and 1st sacral cord segments. These neurons with SPR-LI may receive noxious information from the urinary bladder through the primary afferent fibers with substance P. AU - Lü, Yan AU - Jin, Shan AU - Xu, Tian AU - Qin, Bing AU - Li, Ji AU - Ding, Yu AU - Shigemoto, Ryuichi AU - Mizuno, Noboru ID - 2560 IS - 2 JF - Neuroscience Letters SN - 0304-3940 TI - Expression of c-fos protein in substance P receptor-like immunoreactive neurons in response to noxious stimuli on the urinary bladder: an observation in the lumbosacral cord segments of the rat VL - 198 ER - TY - JOUR AB - The superficial layers of the superior colliculus (SC) have been known to contain many axons showing substance P-like immunoreactivity (SP-LI). We, therefore, immunohistochemically examined the distribution of SP receptor (SPR) in the superficial layers of the SC in the rat by using a specific antibody against SPR. The majority of SC neurons with SPR-LI were distributed in the zonal and the superficial gray layers, the rest of them were in the optic layer. Electron microscopy revealed that SPR-immunoreaction products in SC neurons were distributed not only in postsynaptic sites, but also in non-synaptic regions of perikaryal and dendritic profiles. AU - Ogawa Meguro, Reiko AU - Shigemoto, Ryuichi AU - Itoh, Kazuo AU - Konishi, Akira AU - Mizuno, Noboru ID - 2552 IS - 2 JF - Neuroscience Letters SN - 0304-3940 TI - Immunohistochemical localization of substance P receptor in the superior colliculus. A light and electron microscope study in the rat VL - 166 ER - TY - JOUR AB - Expression patterns of mRNAs of l-AP4-sensitive metabotropic glutamate receptors (mGluR4, mGluR6, mGluR7) in the rat retina were examined by northern blot analysis and in situ hybridization histochemistry. Expression patterns of mGluR4 and mGluR7 mRNAs were quite different from that of mGluR6 mRNA which was expressed at the outer part of the inner nuclear layer. The mGluR4 mRNA was expressed on the cell bodies of the ganglion cells, but not in the inner or outer nuclear layer. The expression of mGluR7 mRNA was observed throughout the entire region of the inner nuclear layer and on the cell bodies of the ganglion cells. AU - Akazawa, Chihiro AU - Ohishi, Hitoshi AU - Nakajima, Yoshiaki AU - Okamoto, Naoyuki AU - Shigemoto, Ryuichi AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2551 IS - 1-2 JF - Neuroscience Letters SN - 0304-3940 TI - Expression of mRNAs of l-AP4-sensitive metabotropic glutamate receptors (mGluR4, mGluR6, mGluR7) in the rat retina VL - 171 ER - TY - JOUR AB - A trpE-fusion protein containing a C-terminal sequence of a rat metabotropic glutamate receptor, mGluR5, was used to produce an antibody. On immunoblot, the antibody specifically reacted with mGluR5 expressed in mammalian cells and rat brain. Immunohistochemical analysis revealed intense mGluR5-like immunoreactivity (LI) in the olfactory bulb, anterior olfactory nuclei, olfactory tubercle, cerebral cortex, hippocampus, lateral septum, striatum, nucleus accumbens, inferior colliculus, and spinal trigeminal nuclei. The distribution pattern of mGluR5-LI corresponds very well with that of mGluR5 mRNA. Electron microscope analysis of the striatum revealed dense accumulation of immunoreaction products in dendrites which were often provided with asymmetrical synapses. These results suggest that mGluR5 is predominantly located in postsynaptic elements. AU - Shigemoto, Ryuichi AU - Nomura, Sakashi AU - Ohishi, Hitoshi AU - Sugihara, Hidemitsu AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2542 IS - 1 JF - Neuroscience Letters SN - 0304-3940 TI - Immunohistochemical localization of a metabotropic glutamate receptor, mGluR5, in the rat brain VL - 163 ER - TY - JOUR AB - A trp E fusion protein containing a C-terminal portion of the rat substance P receptor (SPR) was expressed in bacteria and used to produce an antibody. The antibody specifically reacted with SPR expressed in a mammalian cell line and rat striatum. Light and electron microscope analyses of the rat striatum revealed intense SPR-like immunoreactivity in neuronal somata and dendrites. These immunoreactive neurons constituted ∼ 3% of the total population of striatal neurons; they were putative interneurons of large and medium-sized aspiny types. AU - Shigemoto, Ryuichi AU - Nakaya, Yoshifumi AU - Nomura, Sakashi AU - Ogawa Meguro, Reiko AU - Ohishi, Hitoshi AU - Kaneko, Takeshi AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2541 IS - 2 JF - Neuroscience Letters SN - 0304-3940 TI - Immunocytochemical localization of rat substance P receptor in the striatum VL - 153 ER - TY - JOUR AB - The distribution of NMDA receptor (NMDAR1) on neurons in the peripheral ganglia was examined in the adult rat by in situ hybridization. NMDAR1 mRNA was expressed in all neurons in the sensory and autonomic ganglia examined; in the dorsal root, trigeminal, nodose, superior cervical, and sphenopalatine ganglia. Possible roles of the NMDA receptor on the sensory and autonomic ganglion neurons are discussed. AU - Shigemoto, Ryuichi AU - Ohishi, Hitoshi AU - Nakanishi, Shigetada AU - Mizuno, Noboru ID - 2531 IS - 1-2 JF - Neuroscience Letters SN - 0304-3940 TI - Expression of the mRNA for the rat NMDA receptor (NMDAR1) in the sensory and autonomic ganglion neurons VL - 144 ER - TY - JOUR AB - Injection of large amounts of a mixture of horseradish peroxidase and wheat germ agglutinin-horseradish peroxidase conjugate into the upper cervical segments of the spinal cord in the Japanese monkey (Macaca fuscata) led to the retrograde labeling of a small number of neuronal cell bodies within the rostral part of the subthalamic nucleus of Luys. Direct projection from the subthalamic nucleus to the spinal cord appeared to be much less prominent in the Japanese monkey than in the cat and rat. AU - Mizuno, Noboru AU - Ueyama, Teizo AU - Itoh, Kazuo AU - Satoda, Takahiro AU - Tashiro, Takashi AU - Shigemoto, Ryuichi ID - 2523 IS - 1 JF - Neuroscience Letters SN - 0304-3940 TI - Direct projections from the subthalamic nucleus of Luys to the spinal cord in the Japanese monkey VL - 89 ER -