scholarly journals Molecular cloning and characterization of a novel phospholipase C, PLC-η

2005 ◽  
Vol 389 (1) ◽  
pp. 181-186 ◽  
Author(s):  
Jong-Ik HWANG ◽  
Yong-Seok OH ◽  
Kum-Joo SHIN ◽  
Hyun KIM ◽  
Sung Ho RYU ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Phospholipase C ◽  
Neuronal Cell ◽  
Zona Incerta ◽  
Pleckstrin Homology Domain ◽  
Molecular Biological Analysis ◽  
Splicing Variants ◽  
The Brain

PLC (phospholipase C) plays an important role in intracellular signal transduction by hydrolysing phosphatidylinositol 4,5-bisphosphate, a membrane phospholipid. To date, 12 members of the mammalian PLC isoforms have been identified and classified into five isotypes β, γ, δ, ε and ζ, which are regulated by distinct mechanisms. In the present study, we describe the identification of a novel PLC isoform in the brains of human and mouse, named PLC-η, which contains the conserved pleckstrin homology domain, X and Y domains for catalytic activity and the C2 domain. The first identified gene encoded 1002 (human) or 1003 (mouse) amino acids with an estimated molecular mass of 115 kDa. The purified recombinant PLC-η exhibited Ca2+-dependent catalytic activity on phosphatidylinositol 4,5-bisphosphate. Furthermore, molecular biological analysis revealed that the PLC-η gene was transcribed to several splicing variants. Although some transcripts were detected in most of the tissues we examined, the transcript encoding 115 kDa was restricted to the brain and lung. In addition, the expression of the 115 kDa protein was defined in only nerve tissues such as the brain and spinal cord. In situ hybridization analysis with brain revealed that PLC-η was abundantly expressed in various regions including cerebral cortex, hippocampus, zona incerta and cerebellar Purkinje cell layer, which are neuronal cell-enriched regions. These results suggest that PLC-η may perform fundamental roles in the brain.

scholarly journals Characterization of a Vesivirus Associated with an Outbreak of Acute Hemorrhagic Gastroenteritis in Domestic Dogs

2018 ◽  
Vol 56 (5) ◽  
Author(s):  
Randall W. Renshaw ◽  
Jennifer Griffing ◽  
Jaime Weisman ◽  
Lisa M. Crofton ◽  
Melissa A. Laverack ◽  
...  
Keyword(s):  
Intestinal Tract ◽  
Virus Isolation ◽  
Cho Cell ◽  
Mild Disease ◽  
Reverse Transcription Pcr ◽  
The Brain ◽  
Housing Facility ◽  
Diagnostic Investigations

ABSTRACT Four of eleven affected dogs died despite aggressive treatment during a 2015 focal outbreak of hemorrhagic gastroenteritis following a stay in a pet housing facility. Routine diagnostic investigations failed to identify a specific cause. Virus isolation from fresh necropsy tissues yielded a calicivirus with sequence homology to a vesivirus within the group colloquially known as the vesivirus 2117 strains that were originally identified as contaminants in CHO cell bioreactors. In situ hybridization and reverse transcription-PCR assays of tissues from the four deceased dogs confirmed the presence of canine vesivirus (CaVV) nucleic acids that localized to endothelial cells of arterial and capillary blood vessels. CaVV nucleic acid corresponded to areas of necrosis and hemorrhage primarily in the intestinal tract, but also in the brain of one dog with nonsuppurative meningoencephalitis. This is the first report of an atypical disease association with a putative hypervirulent vesivirus strain in dogs, as all other known strains of CaVV appear to cause nonclinical infections or relatively mild disease. After identification of the CU-296 vesivirus strain from this outbreak, four additional CaVV strains were amplified from unrelated fecal specimens and archived stocks provided by other laboratories. Broader questions include the origins, reservoir(s), and potential for reemergence and spread of these related CaVVs.

scholarly journals Identification and Characterization of Four Autophagy-Related Genes That Are Expressed in Response to Hypoxia in the Brain of the Oriental River Prawn (Macrobrachium nipponense)

2019 ◽  
Vol 20 (8) ◽  
pp. 1856 ◽  
Author(s):  
Shengming Sun ◽  
Ying Wu ◽  
Hongtuo Fu ◽  
Xianping Ge ◽  
Hongzheng You ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Open Reading Frames ◽  
Dependent Manner ◽  
Macrobrachium Nipponense ◽  
Oriental River Prawn ◽  
Adverse Environmental Conditions ◽  
The Brain ◽  
Identification And Characterization

Autophagy is a cytoprotective mechanism triggered in response to adverse environmental conditions. Herein, we investigated the autophagy process in the oriental river prawn (Macrobrachium nipponense) following hypoxia. Full-length cDNAs encoding autophagy-related genes (ATGs) ATG3, ATG4B, ATG5, and ATG9A were cloned, and transcription following hypoxia was explored in different tissues and developmental stages. The ATG3, ATG4B, ATG5, and ATG9A cDNAs include open reading frames encoding proteins of 319, 264, 268, and 828 amino acids, respectively. The four M. nipponense proteins clustered separately from vertebrate homologs in phylogenetic analysis. All four mRNAs were expressed in various tissues, with highest levels in brain and hepatopancreas. Hypoxia up-regulated all four mRNAs in a time-dependent manner. Thus, these genes may contribute to autophagy-based responses against hypoxia in M. nipponense. Biochemical analysis revealed that hypoxia stimulated anaerobic metabolism in the brain tissue. Furthermore, in situ hybridization experiments revealed that ATG4B was mainly expressed in the secretory and astrocyte cells of the brain. Silencing of ATG4B down-regulated ATG8 and decreased cell viability in juvenile prawn brains following hypoxia. Thus, autophagy is an adaptive response protecting against hypoxia in M. nipponense and possibly other crustaceans. Recombinant MnATG4B could interact with recombinant MnATG8, but the GST protein could not bind to MnATG8. These findings provide us with a better understanding of the fundamental mechanisms of autophagy in prawns.

scholarly journals In situ characterization of the brain–microdevice interface using Device Capture Histology

2011 ◽  
Vol 201 (1) ◽  
pp. 67-77 ◽  
Author(s):  
Andrew J. Woolley ◽  
Himanshi A. Desai ◽  
Mitchell A. Steckbeck ◽  
Neil K. Patel ◽  
Kevin J. Otto
Keyword(s):  
In Situ Characterization ◽  
The Brain

Expression and characterization of a pleckstrin homology domain in phospholipase C, PLC-η1

2007 ◽  
Vol 56 (2) ◽  
pp. 247-252 ◽  
Author(s):  
Toru Imai ◽  
Kouki Kasai ◽  
Junichi Kurita ◽  
Kiyoko Fukami ◽  
Mitsuru Tashiro ◽  
...  
Keyword(s):  
Phospholipase C ◽  
Pleckstrin Homology Domain ◽  
Pleckstrin Homology

scholarly journals Chemical probes to potently and selectively inhibit endocannabinoid cellular reuptake

2017 ◽  
Vol 114 (25) ◽  
pp. E5006-E5015 ◽  
Author(s):  
Andrea Chicca ◽  
Simon Nicolussi ◽  
Ruben Bartholomäus ◽  
Martina Blunder ◽  
Alejandro Aparisi Rey ◽  
...  
Keyword(s):  
Membrane Trafficking ◽  
Cannabinoid Receptor ◽  
Facilitated Diffusion ◽  
Chemical Probes ◽  
Analgesic Effects ◽  
Complex Process ◽  
Arachidonoyl Glycerol ◽  
The Brain

The extracellular effects of the endocannabinoids anandamide and 2-arachidonoyl glycerol are terminated by enzymatic hydrolysis after crossing cellular membranes by facilitated diffusion. The lack of potent and selective inhibitors for endocannabinoid transport has prevented the molecular characterization of this process, thus hindering its biochemical investigation and pharmacological exploitation. Here, we report the design, chemical synthesis, and biological profiling of natural product-derivedN-substituted 2,4-dodecadienamides as a selective endocannabinoid uptake inhibitor. The highly potent (IC50= 10 nM) inhibitorN-(3,4-dimethoxyphenyl)ethyl amide (WOBE437) exerted pronounced cannabinoid receptor-dependent anxiolytic, antiinflammatory, and analgesic effects in mice by increasing endocannabinoid levels. A tailored WOBE437-derived diazirine-containing photoaffinity probe (RX-055) irreversibly blocked membrane transport of both endocannabinoids, providing mechanistic insights into this complex process. Moreover, RX-055 exerted site-specific anxiolytic effects on in situ photoactivation in the brain. This study describes suitable inhibitors to target endocannabinoid membrane trafficking and uncovers an alternative endocannabinoid pharmacology.

scholarly journals Phospholipase C from Pseudomonas aeruginosa and Bacillus cereus; characterization of catalytic activity

2014 ◽  
Vol 7 (11) ◽  
pp. 860-866 ◽  
Author(s):  
Nooran Sherif Elleboudy ◽  
Mohammad Mabrouk Aboulwafa ◽  
Nadia Abdel-Haleem Hassouna
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Catalytic Activity ◽  
Bacillus Cereus ◽  
Phospholipase C

In situ characterization of catalytic activity of graphene stabilized small-sized Pd nanoparticles for CO oxidation

2013 ◽  
Vol 283 ◽  
pp. 1076-1079 ◽  
Author(s):  
Bao-Hua Mao ◽  
Chang-Hai Liu ◽  
Xu Gao ◽  
Rui Chang ◽  
Zhi Liu ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Co Oxidation ◽  
Pd Nanoparticles ◽  
In Situ Characterization
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