{"title":"Identification and characterization of a novel Delphilin variant with an alternative N-terminus","authors":"Tetsuji Yamashita , Yohei Miyagi , Michio Ono , Hiroaki Ito , Keiko Watanabe , Tomoko Sonoda , Keisuke Tsuzuki , Seiji Ozawa , Ichiro Aoki , Kenji Okuda , Masayoshi Mishina , Susumu Kawamoto","doi":"10.1016/j.molbrainres.2005.08.006","DOIUrl":"10.1016/j.molbrainres.2005.08.006","url":null,"abstract":"<div><p><span><span><span>Delphilin is identified as a Glutamate receptor δ2 (GluRδ2) subunit interacting </span>protein<span>, consisting of a PDZ domain and formin homology (FH) domains 1 and 2, in addition to a C-terminal coiled-coil structure. Delphilin has been shown to be selectively expressed in cerebellar </span></span>Purkinje cells<span> where it co-localizes with the GluRδ2 subunit at the Purkinje cell-parallel fiber synapses. Although Delphilin specifically interacts with the GluRδ2 C-terminus via its PDZ domain, the physiological role of the interaction is not yet understood. Here, we report that the Delphilin protein exhibits diversity at its N-terminus by variable usage of the first several exons. Interestingly, the two Delphilin mRNAs which correspond to the first one initially identified (now designated as Delphilin α) and the second that contains a newly identified first exon (designated as Delphilin β), show different chronological expression profiles. Delphilin β mRNA was not decreased throughout the cerebellar development in vivo and in vitro, while in vivo Delphilin α mRNA gradually decreases following the first postnatal week. Delphilins α and β also revealed different subcellular distribution with some overlap. Specifically, the cerebellar synaptosomal membrane fraction contained the Delphilin β protein. Both Delphilin α and β localized at the </span></span>dendritic spines with GluRδ2; however, dendritic shafts in cultured Purkinje cells also included Delphilin β. In MDCK cells upon becoming confluent, Delphilin α moved to the cell–cell junction area, whereas Delphilin β maintained a diffuse distribution pattern throughout the cytoplasm. Taken as a whole, these two different Delphilins seemed to play functionally different roles in developing and matured cerebellar Purkinje cells.</p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 83-94"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.08.006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25014174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-11-18Epub Date: 2005-08-24DOI: 10.1016/j.molbrainres.2005.07.012
Ping-An Chang , Rui Chen , Yi-Jun Wu
{"title":"Reduction of neuropathy target esterase does not affect neuronal differentiation, but moderate expression induces neuronal differentiation in human neuroblastoma (SK-N-SH) cell line","authors":"Ping-An Chang , Rui Chen , Yi-Jun Wu","doi":"10.1016/j.molbrainres.2005.07.012","DOIUrl":"10.1016/j.molbrainres.2005.07.012","url":null,"abstract":"<div><p><span>Neuropathy target esterase<span> (NTE) is inhibited and aged by organophosphorus compounds that induce delayed neuropathy in human and some sensitive animals. NTE has been proposed to play a role in neurite outgrowth and process elongation during neurodifferentiation. However, to date, there is no direct evidence of the relevance of NTE in neurodifferentiation under physiological conditions. In this study, we have investigated a possible role for NTE in the all-</span></span><em>trans</em><span><span> retinoic acid-induced differentiation of neuroblastoma cells. The functional inactivation of NTE by RNA interference indicated that reduction of NTE does not affect </span>process outgrowth<span> or differentiation of the cells, although moderate expression of NTE by expression of the NTE esterase domain accelerates the elongation of neurite<span> processes. Mipafox, a neurotoxic organophosphate, was shown to block process outgrowth and differentiation in cells that have lowered NTE activity due to RNA interference, suggesting that mipafox may interact with other molecules to exert its effect in this context.</span></span></span></p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 30-38"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.07.012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24976277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-11-18Epub Date: 2005-09-09DOI: 10.1016/j.molbrainres.2005.08.004
Silke Penschuck , Jesper F. Bastlund , Henrik Sindal Jensen , Tine Bryan Stensbol , Jan Egebjerg , William P. Watson
{"title":"Changes in KCNQ2 immunoreactivity in the amygdala in two rat models of temporal lobe epilepsy","authors":"Silke Penschuck , Jesper F. Bastlund , Henrik Sindal Jensen , Tine Bryan Stensbol , Jan Egebjerg , William P. Watson","doi":"10.1016/j.molbrainres.2005.08.004","DOIUrl":"10.1016/j.molbrainres.2005.08.004","url":null,"abstract":"<div><p>Potassium channels<span><span> containing the KCNQ2 subunit play an important role in the regulation of neuronal excitability<span><span> and therefore have been implicated in epilepsy. This study describes the expression of KCNQ2 subunit immunoreactivity<span> in the basolateral amygdala in two rat models of </span></span>temporal lobe epilepsy<span>, (1) amygdala kindling and (2) spontaneously epileptic rats after </span></span></span>status epilepticus<span><span> induced by hippocampal electrical stimulation. KCNQ2 subunit immunoreactivity was assessed with a commercial antibody raised against a C-terminal part of the KCNQ2 protein. We show that KCNQ2 subunit immunoreactivity is upregulated in the basolateral amygdala in both models and that </span>generalized seizures are required to induce this upregulation. We hypothesize that the upregulation of potassium channels containing the KCNQ2 subunit might represent a mechanism to counteract seizures in experimental temporal lobe epilepsy.</span></span></p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 66-73"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.08.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25002657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Entorhinal deafferentation induces upregulation of SPARC in the mouse hippocampus","authors":"Xin Liu, Guoxin Ying, Wenyuan Wang, Jinghui Dong, Yan Wang, Zimei Ni, Changfu Zhou","doi":"10.1016/j.molbrainres.2005.08.003","DOIUrl":"10.1016/j.molbrainres.2005.08.003","url":null,"abstract":"<div><p><span><span><span>SPARC<span> is a matricellular protein that modulates cell–cell and cell–matrix interactions by virtue of its antiproliferative and counteradhesive properties. Here, we report the denervation-induced upregulation of SPARC mRNA and protein in the mouse hippocampus following transections of the entorhinal afferents. </span></span>Northern blot<span> analysis showed that SPARC mRNA was upregulated in a transient manner in the deafferented mouse hippocampus. In situ hybridization and </span></span>immunohistochemistry<span> confirmed the temporal upregulation of both SPARC mRNA and protein specifically in the denervated areas, which initiated at 7 days postlesion, reached the maximum at 15 as well as 30 days postlesion, and subsided towards normal levels by 60 days postlesion. Double labeling by either a combination of in situ hybridization for SPARC mRNA with immunohistochemistry for glial fibrillary acidic protein<span> or double immunofluorescence staining for both proteins in the hippocampus revealed that SPARC-expressing cells are reactive astrocytes. In respect to the spatiotemporal alterations of SPARC expression in the denervated hippocampus, we suggest that SPARC may be involved in modulation of the denervation-induced plasticity processes such as </span></span></span>glial cell<span> proliferation, axonal sprouting and subsequent synaptogenesis in the hippocampus following entorhinal deafferentation.</span></p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 58-65"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.08.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25279434","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-11-18Epub Date: 2005-09-08DOI: 10.1016/j.molbrainres.2005.08.008
Mihaela Sin , Paul D. Walker , Mohamad Bouhamdan , John P. Quinn , Michael J. Bannon
{"title":"Preferential expression of an AAV-2 construct in NOS-positive interneurons following intrastriatal injection","authors":"Mihaela Sin , Paul D. Walker , Mohamad Bouhamdan , John P. Quinn , Michael J. Bannon","doi":"10.1016/j.molbrainres.2005.08.008","DOIUrl":"10.1016/j.molbrainres.2005.08.008","url":null,"abstract":"<div><p><span>Most CNS studies using recombinant adeno-associated virus type 2 (rAAV-2) vectors have focused on gene delivery for the purpose of gene therapy. In the present study, we examined the feasibility of using rAAV-2 vectors to study the regulation of preprotachykinin-A (PPT-A) promoter activity in striatal medium spiny projection neurons. An rAAV-2 vector incorporating a PPT promoter fragment (shown previously to confer some cell-specificity of expression in vitro) coupled to a green fluorescent protein (GFP) reporter gene was stereotaxically injected into the rat striatum. Since medium spiny projection neurons represent the predominant neuronal type (90–95%) in the striatum, we predicted that the vast majority of GFP-expressing cells would be of this phenotype. Surprisingly, the transgene was actually expressed in a similar number of medium spiny projection neurons and </span>interneurons<span>, while glial expression of GFP was not observed. A preponderance of GFP-expressing interneurons was immunoreactive for the marker neuronal nitric oxide synthase (nNOS). Our results suggest that viral vector-related events that occur during transduction are the determining factor in the pattern of transgene expression observed, while the influence of the transgene promoter appears to be secondary, at least under the conditions employed.</span></p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 74-82"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.08.008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25001735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-11-18Epub Date: 2005-10-13DOI: 10.1016/j.molbrainres.2005.06.005
K. Larivière , M. Samia , A. Lister , G. Van Der Kraak , V.L. Trudeau
{"title":"Sex steroid regulation of brain glutamic acid decarboxylase (GAD) mRNA is season-dependent and sexually dimorphic in the goldfish Carassius auratus","authors":"K. Larivière , M. Samia , A. Lister , G. Van Der Kraak , V.L. Trudeau","doi":"10.1016/j.molbrainres.2005.06.005","DOIUrl":"10.1016/j.molbrainres.2005.06.005","url":null,"abstract":"<div><p><span><span><span><span>GABA, the major inhibitory neurotransmitter of the vertebrate brain, has been shown to play an important role in vertebrate reproduction by regulating </span>LH release and sexual behavior. We have studied the expression of the GABA-synthesizing enzyme, </span>glutamic acid decarboxylase (GAD), in goldfish throughout the </span>reproductive cycle in May (mature), November (early gonadal recrudescence) and February (late gonadal recrudescence) and in response to implanted sex steroids. Levels of GAD</span><sub>67</sub> and GAD<sub>65</sub> mRNA levels in the hypothalamus of both males and females were highest in the early stages of gonadal recrudescence. In the telencephalon, a different seasonal pattern of GAD expression was evident. The telencephalic expression GAD<sub>67</sub>, GAD<sub>65</sub><span><span> and a novel isoform, GAD3, were highest in sexually mature fish in May. Five-day intraperitoneal implantation of gonad-intact fish with </span>testosterone<span> (T), estradiol<span> (E2) or progesterone (P4) did not affect GAD expression in November and February. This is in contrast to results in May when sex differences in steroid responsiveness were evident. Progesterone decreased hypothalamic GAD</span></span></span><sub>67</sub> and GAD<sub>65</sub> in females and was without effect in males. All other treatments did not alter GAD<sub>67</sub>, GAD<sub>65</sub> or GAD3 expression in the hypothalamus. Both T and P4 decreased GAD<sub>67</sub> and GAD<sub>65</sub> levels in the telencephalon of male goldfish but had no effect in females. Serum sex steroid levels in control and implanted mature males and females in May were similar so it is unlikely that sex differences in the GAD responses were a result of differences in serum sex steroid levels. These contrasting effects of sex steroids on males and females suggest important sex differences in the regulation of the GADs in sexually mature goldfish.</p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 1-9"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.06.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25638574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-11-18Epub Date: 2005-11-03DOI: 10.1016/S0169-328X(05)00400-6
{"title":"Brain Research Young Investigator Awards","authors":"","doi":"10.1016/S0169-328X(05)00400-6","DOIUrl":"https://doi.org/10.1016/S0169-328X(05)00400-6","url":null,"abstract":"","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Page v"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0169-328X(05)00400-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"137422141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-11-18Epub Date: 2005-09-13DOI: 10.1016/j.molbrainres.2005.08.005
Joshua G. Hunsberger , Alica H. Bennett , Emmanuel Selvanayagam , Ronald S. Duman , Samuel S. Newton
{"title":"Gene profiling the response to kainic acid induced seizures","authors":"Joshua G. Hunsberger , Alica H. Bennett , Emmanuel Selvanayagam , Ronald S. Duman , Samuel S. Newton","doi":"10.1016/j.molbrainres.2005.08.005","DOIUrl":"10.1016/j.molbrainres.2005.08.005","url":null,"abstract":"<div><p><span>Kainic acid activates non-</span><em>N</em>-methyl-<span>d</span><span>-aspartate (NMDA) glutamate receptors<span><span> where it increases synaptic activity resulting in seizures, neurodegeneration<span><span>, and remodeling. We performed microarray analysis on rat hippocampal tissue following kainic acid treatment in order to study the signaling mechanisms underlying these diverse processes in an attempt to increase our current understanding of mechanisms contributing to such fundamental processes as neuronal protection and neuronal plasticity. The kainic acid-treated rats used in our array experiments demonstrated severe seizure behavior that was also accompanied by neuronal degeneration which is suggested by fluoro-jade B </span>staining and anti-caspase-3 </span></span>immunohistochemistry<span><span>. The gene profile revealed 36 novel kainic acid regulated genes along with additional genes previously reported. The functional roles of these novel genes are discussed. These genes mainly have roles in transcription and to a lesser extent have roles in cell death, extracellular matrix remodeling, </span>cell cycle progression<span><span>, neuroprotection, </span>angiogenesis<span><span>, and synaptic signaling. Gene regulation was confirmed via quantitative real time polymerase chain reaction and </span>in situ hybridization.</span></span></span></span></span></p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"141 1","pages":"Pages 95-112"},"PeriodicalIF":0.0,"publicationDate":"2005-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.08.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25303625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Tetracycline-regulated gene expression in the NSC-34-tTA cell line for investigation of motor neuron diseases","authors":"Elisabetta Babetto , Alessandra Mangolini , Milena Rizzardini , Monica Lupi , Laura Conforti , Paola Rusmini , Angelo Poletti , Lavinia Cantoni","doi":"10.1016/j.molbrainres.2005.07.010","DOIUrl":"10.1016/j.molbrainres.2005.07.010","url":null,"abstract":"<div><p><span>The motor neuron-like cell line NSC-34 has become a widely used in vitro model for motor neuron biology and pathology. We established a tetracycline-regulated gene expression system in this cell line by stably transfecting pTet-Off, which codifies for the </span>tetracycline<span><span> transactivator, the regulatory protein<span> tTA<span><span>. The monoclonal cell lines (NSC-34-tTA) were evaluated for the presence of functional tTA after transient transfection with pBI-EGFP, analyzing the expression of the reporter gene enhanced green fluorescent protein. We evaluated the regulation of tTA function with </span>doxycycline using </span></span></span>fluorescence microscopy and quantitative cytofluorimetric analysis on viable transfected cells. The best-regulated cell line (NSC-34-tTA40) had a 66.4-fold induction for the reporter gene fluorescence in comparison to NSC-34.</span></p><p><span>Alpha-tubulin, GAP-43 and phosphorylated medium and heavy neurofilaments, proteins<span> of importance for the motor neuronal phenotype, were evident in NSC-34-tTA40 by Western blot and </span></span>immunocytochemistry; they were expressed similarly in NSC-34-tTA40 and in NSC-34.</p><p>The cDNA of human Cu/Zn superoxide dismutase, a gene of interest for amyotrophic lateral sclerosis, was cloned into pBI-EGFP, downstream of the tetracycline-responsive bidirectional promoter. This plasmid was transiently transfected into NSC-34-tTA40, and the functionality of bidirectional transcription was verified by determining the expression of enhanced green fluorescent protein and of human Cu/Zn superoxide dismutase. Both proteins were regulated by doxycycline.</p><p>This novel cell line, NSC-34 tTA40, that permits tetracycline-regulated gene expression may prove useful to unravel the mechanisms of motor neuron degeneration.</p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 63-72"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.07.010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25270509","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular Brain ResearchPub Date : 2005-10-31Epub Date: 2005-07-14DOI: 10.1016/j.molbrainres.2005.06.009
Jun Xu
{"title":"Age-related changes in Usp9x protein expression and DNA methylation in mouse brain","authors":"Jun Xu","doi":"10.1016/j.molbrainres.2005.06.009","DOIUrl":"10.1016/j.molbrainres.2005.06.009","url":null,"abstract":"<div><p>Usp9x, a ubiquitin-specific protease implicated in synaptic development, was found to be more abundant in adult as compared to newborn mouse brain tissue. The <em>Usp9x</em><span><span> gene was less methylated in adults than in newborns in both the promoter and the protein coding region. Compared with newborns, the adult mouse brain also had lower levels of Dnmt1, the enzyme responsible for maintaining </span>DNA methylation state. These age-associated changes in DNA methylation and ubiquitin system protein concentrations potentially contribute to developmental changes in brain structure and function.</span></p></div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 17-24"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.06.009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24903404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}