scholarly journals retroLEAP: rAAV-based retrograde trans-synaptic labeling, expression and perturbation

2019 ◽  
Author(s):  
Janine K Reinert ◽  
Ivo Sonntag ◽  
Hannah Sonntag ◽  
Rolf Sprengel ◽  
Patric Pelzer ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Heavy Chain ◽  
Tetanus Toxin ◽  
Brain Regions ◽  
Nervous Systems ◽  
Novel Approach

AbstractIdentifying and manipulating synaptically connected neurons across brain regions remains a core challenge in understanding complex nervous systems. RetroLEAP is a novel approach for retrograde trans-synaptic Labelling, Expression And Perturbation. GFP-dependent recombinase (Flp-DOG) detects trans-synaptic transfer of GFP-tetanus toxin heavy chain fusion protein (GFP-TTC) and activates expression of any gene of interest in synaptically connected cells. RetroLEAP overcomes existing limitations, is non-toxic, highly flexible, efficient, sensitive and easy to implement.

scholarly journals 500. Tetanus Toxin Heavy Chain-Streptoavidin Fusion Protein for Targeted Motor Neuron Gene Delivery through Metabolically Biotinylated AAV

2006 ◽  
Vol 13 ◽  
pp. S193
Author(s):  
Qingshan Teng ◽  
JeffreyS. Bartlett ◽  
Thais Federici ◽  
Jun Yang ◽  
NicholasM. Boulis
Keyword(s):  
Gene Delivery ◽  
Fusion Protein ◽  
Motor Neuron ◽  
Heavy Chain ◽  
Tetanus Toxin

scholarly journals A novel approach for locating mice brain regions of Cryptococcus neoformans CNS invasion

2016 ◽  
Vol 11 ◽  
pp. S136-S143
Author(s):  
Chunting He ◽  
Qingfen Chen ◽  
Longkun Zhu
Keyword(s):  
Motor Cortex ◽  
Cryptococcus Neoformans ◽  
Primary Motor Cortex ◽  
Thalamic Nucleus ◽  
Molecular Layer ◽  
Brain Regions ◽  
Dorsal Lateral Geniculate Nucleus ◽  
Novel Approach ◽  
Stratum Lucidum ◽  
Lateral Posterior

Aim of this study was to locate the brain regions where Cryptococcus interact with brain cells and invade into brain. After 7 days of intratracheal inocula-tion of GFP-tagged Cryptococcus neoformans strains H99, serial cryosections (10 ?m) from 3 C57 BL/6 J mice brains were imaged with immunofluorescence microscopy. GFP-tagged H99 were found in some brain regions such as primary motor cortex-secondary motor cortex, caudate putamen, stratum lucidum of hippocampus, field CA1 of hippocampus, dorsal lateral geniculate nucleus, lateral posterior thalamic nucleus, laterorostral part, lateral posterior thalamic nucleus, mediorostral part, retrosplenial agranular cortex, lateral area of secondary visual cortex, and lacunosum molecular layer of the hippocampus. The results will be very useful for further exploring the mechanism of C. neoformans infection of brain. 

Synthesis

2019 ◽  
pp. 423-472
Author(s):  
Georg F. Striedter ◽  
R. Glenn Northcutt
Keyword(s):  
Functional Organization ◽  
Brain Size ◽  
Brain Regions ◽  
Brain Areas ◽  
Nervous Systems ◽  
Internal Organization ◽  
Vertebrate Brain ◽  
Relative Brain Size

After summarizing the earlier chapters, which focused on the evolution of specific lineages, this chapter examines general patterns in the evolution of vertebrate nervous systems. Most conspicuous is that relative brain size and complexity increased independently in many lineages. The proportional size of individual brain regions tends to change predictably with absolute brain size (and neurogenesis timing), but the scaling rules vary across lineages. Attempts to link variation in the size of individual brain areas (or entire brains) to behavior are complicated in part because the connections, internal organization, and functions of individual brain regions also vary across phylogeny. In addition, major changes in the functional organization of vertebrate brains were caused by the emergence of novel brain regions (e.g., neocortex in mammals and area dorsalis centralis in teleosts) and novel circuits. These innovations significantly modified the “vertebrate brain Bauplan,” but their mechanistic origins and implications require further investigation.

A novel approach to CNS dysfunction of Pompe disease with a fusion protein consisting of anti-transferrin receptor antibody and GAA enzyme

2020 ◽  
Vol 129 (2) ◽  
pp. S150-S151
Author(s):  
Satowa Tanaka ◽  
Aya Yoshioka ◽  
Masafumi Kinoshita ◽  
Sachiho Kida ◽  
Atsushi Imakiire ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Pompe Disease ◽  
Transferrin Receptor ◽  
Receptor Antibody ◽  
Novel Approach ◽  
Cns Dysfunction

ANTIBODY TARGETED FIBRINOLYSIS

1987 ◽  
Author(s):  
Edgar Haber ◽  
Marchall T Runge ◽  
Christoph Bode ◽  
Betsy Branscomb ◽  
Janet Schnee
Keyword(s):  
Monoclonal Antibody ◽  
Fusion Protein ◽  
Cell Line ◽  
Heavy Chain ◽  
Specific Antibody ◽  
Plasminogen Activators ◽  
Amino Terminus ◽  
Beta Chain ◽  
Western Blots ◽  
Competitive Binding Assay

Chemical conjugates of fibrin-specificantibodies and plasminogen activators. Urokinase or tPA were linked covalently toamonoclonal antibody specific for the amino terminus of the beta chain of human fibrin (59D8) by means of the unidirectionalcross-linking reagent SPDP. The fibrinolytic potency of the conjugates at equal amidolytic activities was compared to the native plasminogen activators in an assay measuring lysis of 1251-fibrin monomer covalently linked to Sepharose CL-4B. Urokinase was least potent, tPA exhibited a 10fold increase in fibrinolysis whereas both the urokinase and tPA antibody conjugates and a urokinase-Fab conjugate were 250fold more potent than urokinase and 25 fold more potent than tPA. Enhanced fibrinolysis was fully inhibited by b peptide indicating its dependence on antigen binding. In a plasma assay conjugates of tPA orUK to antibody produced a 3.2- to 4.5-fold enhancement in clot lysis in human plasma over that of the respective unconjugated plasminogen activator. However, the UK-59D8 conjugate was only as potent as tPAalone. Antibody-conjugated tPA or UK consumed less fibrinogen, alpha 2-antiplasminand plasminogen than did the unconjugatedactivators, at equipotent thrombolytic concentrations. In a quantitative rabbit thrombolysis model, the activity of the purified conjugate was compared with that oftPA alone and that of a conjugate betweentPA and a digoxin-specific monoclonal antibody. After correction for spontaneous lysis, tPA-59D8 was shown to be 2.8 to,9.6times more potent than tPA alone. Unconjugated tPA and tPA-digoxin were equipotent.At equivalent thrombolytic concentrations, tPA-59D8 degraded less fibrinogen and consumed less alpha 2-antiplasmin than did tPA alone. These results suggest that tPA can be efficiently directed to the site of a thrombus by conjugation to an antifibrin monoclonal antibody, resulting in both more potent and more selective thrombolysis.A recombinant fusion protein comprising a fibrin-specific antibody site and theB chain of tPA. The rearranged 59D8 heavychain gene was cloned and combined in theexpression vector pSV2gpt withsequence coding for a portion of the Gamma 2b constant region and the catalytic beta chain of t-PA. This construct was transfected into heavy chain loss variant cells derived from the 59D8 hybridoma. Recombinant protein was purified by affinitychromatography and analyzed with Western blots. These revealed a 65-kD heavy chain-t-PA fusion protein that is secreted in association with the 59D8 light chain in the form of a 170-kD disulfide linked dimer. A chromogenic substrate assay showed the fusion protein to have 70 percent of the peptidolytic activity of native t-PA and to activate plasminogen as efficiently as t-PA. In a competitive binding assay, reconstituted antibody was shown to have a binding profile similar to that of native 59D8. Thus by recombinant techniques we have produced a novel hybrid protein capable of high affinity fibrin binding andplasminogen activation.Chemical conjugates between a fibrin-specific and a tPA-specific antibody. A heteroantibody duplex (duplex) with specificities for both tPA and fibrin was synthesized by conjugating iminothiolane-modified anti-tPA monoclonal antibody (TCL8) toantifibrin antibody 59D8. Addition of both duplex and tPA to a plasma clot assay gave more lysis (200 units produced 23.1 lysis; 400 units, 29.5 lysis) than did tPAalone (200 units, 1.8% lysis; 400 units,19% lysis). Despite increased potency associated with duplex addition, fibrinogen and alpha-2-antiplasmin levels at equal tPA concentrations did not differ. Thus, itis possible to concentrate tPA (added separately) to the site of a thrombus in plasma using a heteroantibody duplex with specificities for both tPA and fibrin.Biosynthetically produced heteroduplexantibodies that are both fibin and tPA-specific. The bispecific antibodies were prepared in two ways. First, polyethylene glycol-mediated fusions were performed with two different hybridoma cell lines: anti-fribrin b chain producer, 59D8 and anti-tPA producer, TCL8. TCL8 cells were selected for HPRT-minus variants and then fused with TK-deficient 59D8 cells. One cell line, F36.23, possessed both anti-human fibrin and anti-human t-PA immunoreactivities. A second method yielded another bispecific antibody, F32.1. This cell line was selected after fusing TCL8 (HPRT-minus) cells with spleen cells from a mouseimmunized with a fibrin-like peptide corresponding to the amino terminus of fibrinalpha-chains. Affinity-purified F32.1 andF36.23 retained anti-fibrin and anti-t-PAactivity and enhanced fibrinolytic potency of tPA by a factor of 10.

Development of an improved tetanus vaccine composed of the heavy chain of the tetanus toxin molecule

Toxicon ◽  
1997 ◽  
Vol 35 (4) ◽  
pp. 483
Author(s):  
M. Matsuda ◽  
M. Takahashi ◽  
D. Lei ◽  
N. Sugimoto
Keyword(s):  
Heavy Chain ◽  
Tetanus Toxin ◽  
Toxin Molecule ◽  
Tetanus Vaccine

scholarly journals The inv(16) Fusion Protein Associates with Corepressors via a Smooth Muscle Myosin Heavy-Chain Domain

2003 ◽  
Vol 23 (2) ◽  
pp. 607-619 ◽  
Author(s):  
Kristie L. Durst ◽  
Bart Lutterbach ◽  
Tanawan Kummalue ◽  
Alan D. Friedman ◽  
Scott W. Hiebert
Keyword(s):  
Amino Acids ◽  
Smooth Muscle ◽  
Fusion Protein ◽  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Transcriptional Repression ◽  
Coiled Coil ◽  
Smooth Muscle Myosin ◽  
Muscle Myosin ◽  
Repression Domain

ABSTRACT Inversion(16) is one of the most frequent chromosomal translocations found in acute myeloid leukemia (AML), occurring in over 8% of AML cases. This translocation results in a protein product that fuses the first 165 amino acids of core binding factor β to the coiled-coil region of a smooth muscle myosin heavy chain (CBFβ/SMMHC). CBFβ interacts with AML1 to form a heterodimer that binds DNA; this interaction increases the affinity of AML1 for DNA. The CBFβ/SMMHC fusion protein cooperates with AML1 to repress the transcription of AML1-regulated genes. We show that CBFβ/SMMHC contains a repression domain in the C-terminal 163 amino acids of the SMMHC region that is required for inv(16)-mediated transcriptional repression. This minimal repression domain is sufficient for the association of CBFβ/SMMHC with the mSin3A corepressor. In addition, the inv(16) fusion protein specifically associates with histone deacetylase 8 (HDAC8). inv(16)-mediated repression is sensitive to HDAC inhibitors. We propose a model whereby the inv(16) fusion protein associates with AML1 to convert AML1 into a constitutive transcriptional repressor.

Effects of Chaihu-Guizhi-Ganji on the Levels of Monoamines and Their Related Substances, and Acetylcholine in Discrete Brain Regions of Mice

1996 ◽  
Vol 24 (01) ◽  
pp. 53-64 ◽  
Author(s):  
Tadanobu ltoh ◽  
Seisuke Michijiri ◽  
Shigeo Murai ◽  
Hiroko Saito ◽  
Hiroshi Saito ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Mouse Brain ◽  
Corpus Striatum ◽  
Repeated Administration ◽  
Brain Regions ◽  
Single Administration ◽  
Nervous Systems ◽  
Related Substances

The effects of the extract powder (CggT) from Chaihu-Guizhi-Gajiang-Tang (Saiko-keishi-kankyo-to, in Japanese) on the monoamines and their related substances and the acetylcholine in mouse brain were examined. 1) A single administration of CggT significantly increased the levels of HVA and 5-HIAA in the cerebral cortex, hypothalamus, corpus striatum and hippocampus at 75 mg/kg, and those in the hypothalamus, corpus striatum and hippocampus at 750 mg/kg. 2) The repeated administration of CggT significantly increased the level of 5-HT in the hippocampus at 75 mg/kg, and the levels of 5-HT in the corpus striatum and of NE and 5-HT in the hippocampus at 750 mg/kg. 3) The level of ACh was significantly increased in the hypothalamus alone after single administration of CggT. These findings suggest that CggT stimulates function of the dopaminergic and serotonergic nervous systems in mice, but not most of the NEnergic and cholinergic nervous systems.

HC fragment (C-terminal portion of the heavy chain) of tetanus toxin activates protein kinase C isoforms and phosphoproteins involved in signal transduction

2001 ◽  
Vol 356 (1) ◽  
pp. 97-103 ◽  
Author(s):  
Carles GIL ◽  
Imane CHAIB-OUKADOUR ◽  
Juan BLASI ◽  
José AGUILERA
Keyword(s):  
Signal Transduction ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Tyrosine Phosphorylation ◽  
Heavy Chain ◽  
Tetanus Toxin ◽  
Signal Transduction Pathways ◽  
Terminal Portion ◽  
Kinase C ◽  
Pkc Isoforms

A recent report [Gil, Chaib-Oukadour, Pelliccioni and Aguilera (2000) FEBS Lett. 481, 177–182] describes activation of signal transduction pathways by tetanus toxin (TeTx), a Zn2+-dependent endopeptidase synthesized by the Clostridium tetani bacillus, which is responsible for tetanus disease. In the present work, specific activation of protein kinase C (PKC) isoforms and of intracellular signal-transduction pathways, which include nerve-growth-factor (NGF) receptor trkA, phospholipase C(PLC)γ-1 and extracellular regulated kinases (ERKs) 1 and 2, by the recombinant C-terminal portion of the TeTx heavy chain (HC-TeTx) is reported. The activation of PKC isoforms was assessed through their translocation from the soluble (cytosolic) compartment to the membranous compartment, showing that clear translocation of PKC-α, −β, −γ and −δ isoforms exists, whereas PKC-∊ showed a slight decrease in its soluble fraction immunoreactivity. The PKC-∊ isoform showed no consistent response. Using immunoprecipitation assays against phosphotyrosine residues, time- and dose-dependent increases in tyrosine phosphorylation were observed in the trkA receptor, PLCγ-1 and ERK-1/2. The effects shown by the HC-TeTx fragment on tyrosine phosphorylation were compared with the effects produced by NGF. The trkA and ERK-1/2 activation were corroborated using phospho-specific antibodies against trkA phosphorylated on Tyr490, and antibodies against Thr/Tyr phosphorylated ERK-1/2. Moreover, PLCγ-1 phosphorylation was supported by its HC-TeTx-induced translocation to the membranous compartment, an event related to PLCγ-1 activation. Since HC-TeTx is the domain responsible for membrane binding and lacks catalytic activity, the activations described here must be exclusively triggered by the interaction of TeTx with a membrane component.

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