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• {"link"=>{"url"=>"http://www.citeulike.org/user/scarley/article/11483517"}, "post_time"=>"2012-10-17 06:27:49", "tag"=>["activity-genetics-physiology", "amyotrophic", "animals", "bodies-metabolism", "cognition-physiology", "degeneration-genetics-metabolism-pathology-physiopathology", "dna-binding", "drosophila", "expression", "female", "frontotemporal", "gene", "humans", "junction-cytology-pathology", "knockdown", "larva-genetics", "lateral", "learning-physiology", "lobar", "male", "melanogaster-cytology-genetics-metabolism-physiology", "motor", "mushroom", "mutation", "neuromuscular", "neurons-cytology-metabolism-pathology", "organ", "phenotype", "presynaptic", "proteins-deficiency-genetics-metabolism", "regulation", "sclerosis-genetics-metabolism-pathology-physiopathology", "specificity", "techniques", "terminals-metabolism"], "linkout"=>[{"type"=>"URL", "url"=>"http://www.ncbi.nlm.nih.gov/pubmed/21673800"}, {"type"=>"DOI", "url"=>"http://dx.doi.org/10.1371/journal.pone.0020371"}], "username"=>"scarley", "article_id"=>"11483517"}

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{"title"=>"Neuronal function and dysfunction of drosophila dTDP", "type"=>"journal", "authors"=>[{"first_name"=>"Meng Jau", "last_name"=>"Lin", "scopus_author_id"=>"37042007600"}, {"first_name"=>"Ching Wei", "last_name"=>"Cheng", "scopus_author_id"=>"56309736900"}, {"first_name"=>"C. K James", "last_name"=>"Shen", "scopus_author_id"=>"7402860269"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-79957801387", "pui"=>"361862764", "doi"=>"10.1371/journal.pone.0020371", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"79957801387", "pmid"=>"21673800"}, "id"=>"e0e6c697-de52-3db6-be83-261f0da6b598", "abstract"=>"BACKGROUND: TDP-43 is an RNA- and DNA-binding protein well conserved in animals including the mammals, Drosophila, and C. elegans. In mammals, the multi-function TDP-43 encoded by the TARDBP gene is a signature protein of the ubiquitin-positive inclusions (UBIs) in the diseased neuronal/glial cells of a range of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-U).\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: We have studied the function and dysfunction of the Drosophila ortholog of the mammalian TARDBP gene, dTDP, by genetic, behavioral, molecular, and cytological analyses. It was found that depletion of dTDP expression caused locomotion defect accompanied with an increase of the number of boutons at the neuromuscular junctions (NMJ). These phenotypes could be rescued by overexpression of Drosophila dTDP in the motor neurons. In contrast, overexpression of dTDP in the motor neurons also resulted in reduced larval and adult locomotor activities, but this was accompanied by a decrease of the number of boutons and axon branches at NMJ. Significantly, constitutive overexpression of dTDP in the mushroom bodies caused smaller axonal lobes as well as severe learning deficiency. On the other hand, constitutive mushroom body-specific knockdown of dTDP expression did not affect the structure of the mushroom bodies, but it impaired the learning ability of the flies, albeit moderately. Overexpression of dTDP also led to the formation of cytosolic dTDP (+) aggregates.\\n\\nCONCLUSION/SIGNIFICANCE: These data together demonstrate the neuronal functions of dTDP, and by implication the mammalian TDP-43, in learning and locomotion. The effects of mis-expression of dTDP on Drosophila NMJ suggest that eukaryotic TDP-43 guards against over development of the synapses. The conservation of the regulatory pathways of functions and dysfunctions of Drosophila dTDP and mammalian TDP-43 also shows the feasibility of using the flies as a model system for studying the normal TDP-43 function and TDP-43 proteinopathies in the vertebrates including human.", "link"=>"http://www.mendeley.com/research/neuronal-function-dysfunction-drosophila-dtdp", "reader_count"=>52, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Student > Doctoral Student"=>5, "Researcher"=>16, "Student > Ph. D. Student"=>15, "Student > Master"=>3, "Other"=>3, "Student > Bachelor"=>4, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Student > Doctoral Student"=>5, "Researcher"=>16, "Student > Ph. D. Student"=>15, "Student > Master"=>3, "Other"=>3, "Student > Bachelor"=>4, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Nursing and Health Professions"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>30, "Medicine and Dentistry"=>11, "Neuroscience"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>11}, "Neuroscience"=>{"Neuroscience"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>30}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Japan"=>1, "China"=>1, "United Kingdom"=>3, "Italy"=>1}, "group_count"=>1}

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• {"files"=>["https://ndownloader.figshare.com/files/768761"], "description"=>"<p>(A) Organization of the six-exon <i>dTDP</i> gene in the <i>dTDP^ex26</i> line. This semi-lethal <i>dTDP</i> was generated by imprecise excision of the P-element KG08578 leading to a 932 bp-deletion (the hatched block) in the 5′ end of the <i>dTDP</i> gene. The primers used for PCR in the RT-PCR analysis are indicated. Also shown is the most neighboring gene CG4585 located at upstream of dTDP. Lower panels, RT-PCR analysis of <i>dTDP</i> gene expression in the wild type (<i>yw</i>, left panel) and homozygous <i>dTDP^ex26</i> (right panel). Note the absence of dTDP signal in the <i>dTDP^ex26</i> samples. (B) Immunostaining of the third instar larval segment A3 muscle 6/7 (a,b) and the third instar larval ventral nerve cords (c,d) from <i>yw</i> control and <i>dTDP^ex26</i> mutant with anti-dTDP antibody (green). The nuclei were visualized by DAPI (blue), and the actin cytoskeleton was labeled by phalloidin (magenta). The neurons of the ventral nerve cords were also stained with anti-elav (red). Panel rows a and c show the nuclear localization of dTDP in the <i>yw</i> control and panel rows b and d show the lack of anti-dTDP signals in the <i>dTDP^ex26</i> mutant. (C) Larval movement assay. The movement of the wandering third instar larvae on the agar plate was monitored for 2 minutes and the average distance of the larval trails was calculated. Note the severe movement defect of the homozygous <i>dTDP^ex26</i> mutant in comparison to the <i>yw</i> control, and the partial rescue of this phenotype in <i>dTDP^ex26</i>;<i> D42>dTDP#18-1</i> (<i>N</i> = 15, ***, <i>p</i><0.0001). (D) Comparison of the bouton number/muscle area of <i>yw</i>, <i>dTDP^ex26</i> and <i>dTDP^ex26; D42>dTDP#18-1</i>. The A3 NMJs on muscles 6/7 of the late third instar larvae were co-stained with antibodies against HRP (red) and Synapsin (green), respectively, as exemplified in the right 2 panels. The <i>dTDP^ex26</i> mutant had higher bouton number per unit area of the muscle than the <i>yw</i> control, and this phenotype of <i>dTDP^ex26</i> was rescued in <i>dTDP^ex26; D42>dTDP#18-1</i>. *, <i>p</i><0.05. The averages of the total bouton numbers are: <i>yw</i>, 67±17 (N = 18); <i>dTDP^ex26</i>, 94±14 (N = 16); <i>dTDP^ex26</i>;<i> D42>dTDP#18-1</i>, 75±8 (N = 12). The means of the muscle areas are: <i>yw</i>, 63,187 µm²; <i>dTDP^ex26</i>, 62,565 µm²; <i>dTDP^ex26; D42>dTDP#18-1</i>, 64,310 µm².</p>", "links"=>[], "tags"=>["imprecise", "excision", "mutant"], "article_id"=>439129, "categories"=>["Physiology", "Cell Biology", "Neuroscience", "Genetics"], "users"=>["Meng-Jau Lin", "Ching-Wei Cheng", "C.-K. James Shen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020371.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characteristics_of_the_dTDP_imprecise_excision_mutant_dTDP_ex26_/439129", "title"=>"Characteristics of the <i>dTDP</i> imprecise excision mutant <i>dTDP^ex26</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:50:36"}
• {"files"=>["https://ndownloader.figshare.com/files/769019"], "description"=>"<p>The performance indexes of the control <i>OK107>+</i> flies and flies with mushroom body-specific knockdown (<i>OK107>38377</i> and <i>OK107>38379</i>) or overexpression (<i>OK107>dTDP#5-1 and OK107>dTDP#18-1</i>) of dTDP were measured by the olfactory learning tests. Note the dramatic lowering of the indexes of the <i>dTDP</i>-overexpressing lines (***, <i>p</i><0.0001) and the relatively small but significant lowering of the index of <i>OK107>38377</i> knockdown line (*, <i>p</i><0.05). All value represented mean ± SD. <i>N</i> = 8 performance indexes per group.</p>", "links"=>[], "tags"=>["knockdown", "overexpressing"], "article_id"=>439391, "categories"=>["Physiology", "Cell Biology", "Neuroscience", "Genetics"], "users"=>["Meng-Jau Lin", "Ching-Wei Cheng", "C.-K. James Shen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020371.g003", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Learning_tests_of_Drosophila_dTDP_knockdown_and_overexpressing_flies_/439391", "title"=>"Learning tests of <i>Drosophila dTDP</i> knockdown and overexpressing flies.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:52:05"}
• {"files"=>["https://ndownloader.figshare.com/files/769336"], "description"=>"<p>(A) Whole-mount immunostaining analysis of the larval ventral nerve cords. (a), control flies <i>D42>+</i>; (b) <i>D42>dTDP#18-1</i>; (c) <i>D42>dTDP#5-1</i>; (d) magnified pictures of the boxed area in (c). Red, dTDP; blue, DAPI; green, mCD8::GFP. Note the predominant nuclear localization of dTDP in (a). (b), in the low-level <i>dTDP</i>-overexpressing flies (<i>D42>dTDP#18-1</i>), dTDP was distributed in both the nucleus and cytosol. (c) and (d), in the high-level <i>dTDP</i>-overexpressing flies (<i>D42>dTDP#5-1</i>), dTDP became predominantly localized in the cytosol (arrows in d) and often formed dTDP (+) aggregates (arrowheads in d). (B) Histograph of the densities of the bouton numbers. NMJ staining of the third instar larvae was carried out for <i>D42>+</i>, <i>D42>dTDP#5-1</i>, and <i>D42>dTDP#18-1</i>, and the densities of the bouton numbers were counted and compared. Note that a decrease of the bouton number was observed only in the high-level <i>dTDP</i>-overexpressing fly line (<i>D42>dTDP#5-1</i>), **, <i>p</i><0.001. The averages of the total bouton numbers are: <i>D42>+</i>, 80±4 (N = 20); <i>D42>dTDP#5-1</i>, 46±15 (N = 12); <i>D42>dTDP#18-1</i>, 74±17 (N = 20). The means of the muscle areas are: <i>D42>+</i>, 75,925 µm²; <i>D42>dTDP#5-1</i>, 56,177 µm²; <i>D42>dTDP#18-1</i>, 71,189 µm². (C) Larvae movement assay. Parallel to data of (B), only the larvae with high dTDP overexpression (<i>D42>dTDP#5-1</i>) showed severe movement defect. (D) and (E) Pathogenesis of adult flies with motor neuron-specific overexpression of dTDP. (D) Movement defect of fly line <i>D42>dTDP#18-1</i>. Phototaxis was performed at three different ages of the adult flies and the movement indexes were normalized to that of the control line <i>D42>+</i>. Note the age-dependent declines of the performance of the <i>dTDP</i>-overexpressing flies. <i>N</i> = 5, **, <i>p</i><0.001. (E) Whole-mount immunostaining of the thoracic ganglia of day-9 adult flies. Red, dTDP; blue, DAPI; green, mCD8::GFP. (a) Control flies (<i>D42>+</i>). Note the predominant nuclear localization of dTDP. (b) <i>D42>dTDP#18-1</i> flies. Two regions under the confocal microscope are shown. Note the presence of cells with mainly cytoplasmic distribution of dTDP, as exemplified by cells pointed with the arrows. The dTDP (+) aggregates in the cytoplasm are exemplified by the arrowheads. (F) Western blotting analysis of soluble and insoluble proteins in the heads and thoraces of 13-day old <i>D42>dTDP#18-1</i> flies with the motor neuron-specific overexpression of dTDP. Different fractions of the protein extracts were isolated from the heads and thoraces as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020371#s4\" target=\"_blank\">Materials and Methods</a>, and analyzed by Western blotting with use of anti-dTDP and anti-tubulin. T, total protein; R, RIPA-soluble fraction; U, urea-soluble fraction. Note the presence of dTDP in the urea-soluble fraction from <i>D42>dTDP#18-1</i> but not <i>D42>+</i>.</p>", "links"=>[], "tags"=>["flies", "neuron-specific", "overexpression"], "article_id"=>439707, "categories"=>["Physiology", "Cell Biology", "Neuroscience", "Genetics"], "users"=>["Meng-Jau Lin", "Ching-Wei Cheng", "C.-K. James Shen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020371.g005", "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pathologies_of_flies_with_motor_neuron_specific_overexpression_of_dTDP_/439707", "title"=>"Pathologies of flies with motor neuron-specific overexpression of dTDP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:53:55"}
• {"files"=>["https://ndownloader.figshare.com/files/769179"], "description"=>"<p>(A) Lower locomotive activity of larvae (<i>elav>38377</i>) with knockdown of dTDP than the control (<i>elav>+</i>) (N = 15, *, <i>p</i><0.05). (B) Quantitative comparison of NMJ bouton numbers of <i>elav>+</i> and <i>elav>38377</i> larvae after normalization to the total muscle 6/7 areas. Note the higher bouton density of the knockdown larvae (<i>elav>38377</i>) than the control (<i>elav>+</i>). *, <i>p</i><0.05. The averages of the total bouton numbers are: <i>elav>+</i>, 95±14; <i>elav>38377</i>, 117±14. The means of the muscle areas are: <i>elav>+</i>, 69,937 µm²; <i>elav>38377</i>, 65,166 µm². (N = 12 in all cases). (C) Whole-mount immunostaining analysis of the larval ventral nerve cords with <i>dTDP</i>-knockdown in the pan-neurons. (a), control flies <i>elav>+</i>; (b) <i>elav>38377</i>. Red, dTDP; blue, DAPI; green, mCD8::GFP. Note that the pan-neuron-driven dsRNA did not completely knock down the dTDP expression since there were still some dTDP signals in the larval ventral nerve cords of <i>elav>38377</i>.</p>", "links"=>[], "tags"=>["locomotive", "nmj", "staining", "larvae", "knockdown"], "article_id"=>439549, "categories"=>["Physiology", "Cell Biology", "Neuroscience", "Genetics"], "users"=>["Meng-Jau Lin", "Ching-Wei Cheng", "C.-K. James Shen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020371.g004", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_locomotive_activity_and_NMJ_staining_of_larvae_with_dTDP_knockdown_in_the_pan_neurons_/439549", "title"=>"The locomotive activity and NMJ staining of larvae with <i>dTDP</i> knockdown in the pan-neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:53:00"}
• {"files"=>["https://ndownloader.figshare.com/files/768896"], "description"=>"<p>(A) Distribution of the endogenous dTDP proteins in the mushroom bodies of the <i>OK107>+</i> flies as analyzed by immunostaining with anti-dTDP (red), DAPI (blue), and mCD8::GFP (green). (a) a low-magnification photo; (b), higher magnification photo of the boxed area in (a); (c), higher magnification photo of the boxed area in (b). Note the predominant nuclear localization of dTDP. (B) Western blot of the total head protein extracts from the adult flies. The blot was probed with anti-dTDP and anti-tubulin. Note the decreases, by approximately 46% and 23%, of the dTDP levels in the heads of <i>OK107>38377</i> (lane 2) and <i>OK107>38379</i> (lane 3), respectively, with mushroom body-specific dsRNA-knockdown of dTDP. Also, dTDP levels in the <i>dTDP</i>-overexpressing lines <i>OK107>dTDP#5-1</i> (lane 4) and <i>OK107>dTDP#18-1</i> (lane 5) were 7- and 3 fold, respectively, higher than the <i>OK107>+</i> control (lane 1). (C) GFP-lighted morphologies of the mushroom bodies in <i>OK107>+</i> (a), <i>OK107>38377</i> (b), <i>OK107>dTDP#5-1</i> (c), and <i>OK107>dTDP#18-1</i> (d). The α, α′, β, β′, and γ lobes were indicated in (a). Note the smaller lobes in (c) and (d). (D) Immunostaining of dTDP in the brains of <i>OK107>38377</i> flies with mushroom body-specific knockdown of dTDP expression. Note the significantly reduced signals of dTDP in the mushroom bodies when compared to the <i>yw</i> control in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020371#pone-0020371-g002\" target=\"_blank\">Fig. 2A-a</a>. (E) Subcellular distribution of dTDP in the mushroom bodies of the <i>dTDP</i>-overexpressing line <i>OK107>dTDP#5-1</i>. (a) a set of low magnification pictures; (b) and (c), high magnification pictures of the immunostaining patterns of the <i>OK107>dTDP#5-1</i> mushroom body, with (b) from the boxed area in (a) and (c) from the boxed area in (b). Note the presence of cells with mainly cytoplasmic dTDP (the arrow in b) and cells with cytoplasmic dTDP (+) aggregates (the arrowheads). (F) Western blotting analysis of soluble and insoluble proteins in the heads of 3-day old <i>dTDP</i>-overexpressing flies. Different fractions of protein extracts were isolated from 3 days-old fly heads as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020371#s4\" target=\"_blank\">Materials and Methods</a>, and analyzed by Western blotting with use of anti-dTDP and anti-tubulin. Note the presence of high molecular weight dTDP species in the urea-soluble fraction of <i>OK107>dTDP#5-1</i>. T, total protein; R, RIPA-soluble fraction; U, urea-soluble fraction.</p>", "links"=>[], "tags"=>["brains", "3-day", "mutant"], "article_id"=>439271, "categories"=>["Physiology", "Cell Biology", "Neuroscience", "Genetics"], "users"=>["Meng-Jau Lin", "Ching-Wei Cheng", "C.-K. James Shen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020371.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_dTDP_in_the_adult_brains_of_3_day_old_wild_type_and_mutant_Drosophila_/439271", "title"=>"dTDP in the adult brains of 3-day old wild type and mutant <i>Drosophila</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:51:24"}

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F1000Prime