scholarly journals The Drosophila slamdance Gene: A Mutation in an Aminopeptidase Can Cause Seizure, Paralysis and Neuronal Failure

Genetics ◽  
2002 ◽  
Vol 162 (3) ◽  
pp. 1283-1299 ◽  
Author(s):  
HaiGuang Zhang ◽  
Jeff Tan ◽  
Elaine Reynolds ◽  
Daniel Kuebler ◽  
Sally Faulhaber ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Seizure Threshold ◽  
Electrical Shock ◽  
Hyperactive Behavior ◽  
Mrna In Situ Hybridization ◽  
Lacz Staining ◽  
Drosophila Homolog ◽  
The Central Nervous System

Abstract We report here the characterization of slamdance (sda), a Drosophila melanogaster “bang-sensitive” (BS) paralytic mutant. This mutant exhibits hyperactive behavior and paralysis following a mechanical “bang” or electrical shock. Electrophysiological analyses have shown that this mutant is much more prone to seizure episodes than normal flies because it has a drastically lowered seizure threshold. Through genetic mapping, molecular cloning, and RNA interference, we have demonstrated that the sda phenotype can be attributed to a mutation in the Drosophila homolog of the human aminopeptidase N (APN) gene. Furthermore, using mRNA in situ hybridization and LacZ staining, we have found that the sda gene is expressed specifically in the central nervous system at particular developmental stages. Together, these results suggest that the bang sensitivity in sda mutants is caused by a defective APN gene that somehow increases seizure susceptibility. Finally, by using the sda mutation as a sensitized background, we have been able to identify a rich variety of sda enhancers and other independent BS mutations.

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 Characterization of Putative Effectors from the Cereal Cyst Nematode Heterodera avenae

2018 ◽  
Vol 108 (2) ◽  
pp. 264-274 ◽  
Author(s):  
Jiang-Kuan Cui ◽  
Huan Peng ◽  
Fen Qiao ◽  
Gao-Feng Wang ◽  
Wen-Kun Huang ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Target Genes ◽  
Developmental Stages ◽  
Heterodera Avenae ◽  
Parasitic Nematodes ◽  
Sequencing Analysis ◽  
Rnai Technology

Few molecular details of effectors of Heterodera avenae parasitism are known. We performed a high-throughput sequencing analysis of the H. avenae transcriptome at five developmental stages. A total of 82,549 unigenes were ultimately obtained, and 747 transcripts showed best hits to genes putatively encoding carbohydrate-active enzymes in plant-parasitic nematodes that play an important role in the invasion process. A total of 1,480 unigenes were homologous to known phytonematode effectors, and 63 putative novel effectors were identified in the H. avenae transcriptomes. Twenty-three unigenes were analyzed by qRT-PCR and confirmed to be highly expressed during at least one developmental stage. For in situ hybridization, 17 of the 22 tested putative effectors were specifically expressed and located in the subventral gland cells, and five putative novel effectors were specifically expressed in the dorsal gland. Furthermore, 115 transcripts were found to have putative lethal RNA interference (RNAi) phenotypes. Three target genes with lethal RNAi phenotypes and two of the four tested putative effectors were associated with a decrease in the number of cysts through in vitro RNAi technology. These transcriptomic data lay a foundation for further studies of interactions of H. avenae with cereal and H. avenae parasitic control.

Cerebral Ovine Herpesvirus 2 Infection of Cattle Is Associated With a Variable Neuropathological Phenotype

2020 ◽  
pp. 030098582097049
Author(s):  
Melanie M. Hierweger ◽  
Céline L. Boujon ◽  
Ronja V. Kauer ◽  
Mireille Meylan ◽  
Torsten Seuberlich ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Viral Encephalitis ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
The Central Nervous System ◽  
Consistent Feature ◽  
Polymerase Chain ◽  
The Brain

Cross-species infection with ovine herpesvirus 2 (OvHV-2) in cattle causes malignant catarrhal fever (MCF). MCF may involve the central nervous system (CNS) with necrotizing arteritis and/or vasculitis described to be unique to MCF and discriminatory compared to other viral CNS infections. However, a systematic histopathological characterization of the neural form of MCF in cattle is lacking. We examined medulla oblongata ( n = 9) or the entire brain ( n = 9) of 18 cattle in which OvHV-2 was identified by quantitative polymerase chain reaction (qPCR), in order to pinpoint potential variations in neuropathology. In 2/18 animals (11%) no lesions were identified, while 16/18 cattle (89%) had brain lesions of varying severity. Presence and quantities of OvHV-2 nucleic acid were determined by in situ hybridization and qPCR, respectively, and were related to the severity of lesions. Fifteen of 18 animals (83%) showed vasculitis, which was mainly of the lymphohistiocytic type, while pathognomonic necrotizing arteritis was only rarely present. Neuroparenchymal lesions included gliosis and/or neuronal changes in 7/16 brains with lesions (44%). The number of CD3+ lymphocytes was highest in animals with simultaneous vascular and neuroparenchymal lesions and high viral genome load. In one animal, OvHV-2 was exclusively observed in CD3+ lymphocytes but not in neurons or microglia. In conclusion, the neuropathological phenotype of bovine MCF in the brain was variable. In some cases, lesions mimicked neurotropic viral encephalitis, while pathognomonic necrotizing arteritis was not a consistent feature of neural MCF. Therefore, molecular detection of OvHV-2 is warranted in the presence of nonsuppurative encephalitis and in the absence of necrotizing arteritis.

Neurocartography: Geometry and Biology of the Central Nervous System

1997 ◽  
Vol 3 (S2) ◽  
pp. 295-296
Author(s):  
O. J. Tretiak ◽  
J. Nissanov
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Brain Function ◽  
Spatial Organization ◽  
Mrna In Situ Hybridization ◽  
Functional Images ◽  
Recent Developments ◽  
The Central Nervous System ◽  
And Function

The central nervous system of a vertebrate organism exhibits a very complex spatial organization structure and function. These relationships are the subject of intense study for over a century, and recent developments in imaging have attracted ever increasing effort devoted to the understanding of brain function. One can produce any number of quantitative images that provide maps of the anatomy and function of nerve tissues. For example, autoradiography can yield functional images (2-deoxy glucose), maps of neurotransmitters receptors (over 100 know types), and gene expression labeled with complementary mRNA (in-situ hybridization). Immunohistochemistry produces maps of a large variety of neuroactive components, such as transmitters.To illustrate a typical procedure, we describe the mapping of brain function with 2-deoxy glucose (2DG). A rat performing some task is injected with a solution of 2DG radiolabeled with 14C¨ Subsequently, the animal is sacrificed, the brain is cryosectioned (ca. 20 μm), and contact autoradio-grams of these sections are made on X-ray film.

scholarly journals Isolation and functional characterization of two dioxygenasese putatively involved in bixin biosynthesis in annatto (Bixa orellana L.)

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7064 ◽  
Author(s):  
Victor Manuel Carballo-Uicab ◽  
Yair Cárdenas-Conejo ◽  
Alba Adriana Vallejo-Cardona ◽  
Margarita Aguilar-Espinosa ◽  
Jacobo Rodríguez-Campos ◽  
...  
Keyword(s):  
Food Industry ◽  
Developmental Stages ◽  
Functional Characterization ◽  
Economic Value ◽  
Bixa Orellana ◽  
Reverse Transcription Pcr ◽  
Bona Fide

Carotenoid cleavage dioxygenases (CCDs) are enzymes that have been implicated in the biosynthesis of a wide diversity of secondary metabolites with important economic value, including bixin. Bixin is the second most used pigment in the world’s food industry worldwide, and its main source is the aril of achiote (Bixa orellana L.) seeds. A recent transcriptome analysis of B. orellana identified a new set of eight CCD members (BoCCD4s and BoCCD1s) potentially involved in bixin synthesis. We used several approaches in order to discriminate the best candidates with CCDs genes. A reverse transcription-PCR (RT-qPCR) expression analysis was carried out in five developmental stages of two accessions of B. orellana seeds with different bixin contents: (P13W, low bixin producer and N4P, high bixin producer). The results showed that three BoCCDs (BoCCD4-1, BoCCD4-3, and BoCCD1-1) had an expression pattern consistent with bixin accumulation during seed development. Additionally, an alignment of the CCD enzyme family and homology models of proteins were generated to verify whether the newly proposed CCD enzymes were bona fide CCDs. The study confirmed that these three enzymes were well-preserved and belonged to the CCD family. In a second selection round, the three CCD genes were analyzed by in situ RT-qPCR in seed tissue. Results indicated that BoCCD4-3 and BoCCD1-1 exhibited tissue-specific expressions in the seed aril. To test whether the two selected CCDs had enzymatic activity, they were expressed in Escherichia coli; activity was determined by identifying their products in the crude extract using UHPLC-ESI-QTOF-MS/MS. The cleavage product (bixin aldehyde) was also analyzed by Fourier transform infrared. The results indicated that both BoCCD4-3 and BoCCD1-1 cleave lycopene in vitro at 5,6-5′,6′.

SERS characterization of dopamine and in situ dopamine polymerization on silver nanoparticles

2021 ◽  
Author(s):  
Isidro Badillo-Ramírez ◽  
Jose Saniger ◽  
Juergen Popp ◽  
Dana Cialla-May
Keyword(s):  
Central Nervous System ◽  
Silver Nanoparticles ◽  
Nervous System ◽  
Psychological Disorder ◽  
The Central Nervous System ◽  
Analytical Approaches

Dopamine (DA) regulates several functions in the central nervous system and its depletion is responsible for psychological disorder like Parkinson´s disease. Several analytical approaches have been presented for DA detection...

scholarly journals Functional characterization of the biogenic amine transporter system on human macrophages

2021 ◽  
Author(s):  
Phillip M Mackie ◽  
Adithya Gopinath ◽  
Dominic M Montas ◽  
Alyssa Nielsen ◽  
Rachel Nolan ◽  
...  
Keyword(s):  
Biogenic Amine ◽  
Functional Characterization ◽  
Human Monocyte ◽  
Paracrine Signaling ◽  
Macrophage Response ◽  
Reverse Transport ◽  
The Central Nervous System ◽  
Health And Disease

AbstractMonocyte-derived macrophages are key players in tissue homeostasis and disease regulated by a variety of signaling molecules. Recent literature has highlighted the ability for biogenic amines to regulate macrophage functions, but the mechanisms governing biogenic amine signaling on and around immune cells remains nebulous. In the central nervous system, biogenic amine transporters are regarded as the master regulators of neurotransmitter signaling. While we and others have shown macrophages express these transporters, relatively little is known of their function on these cells. To address these knowledge gaps, we interrogated the function of norepinephrine (NET) and dopamine (DAT) transporters on human monocyte-derived macrophages. We found that both NET and DAT are present and can uptake substrate from the extracellular space at baseline. Not only was DAT expressed in cultured macrophages, but it was also detected in a subset of intestinal macrophages in situ. Surprisingly, we discovered a NET-independent, DAT-mediated immuno-modulatory mechanism in response to lipopolysaccharide (LPS). LPS induced reverse transport of dopamine through DAT, engaging autocrine/paracrine signaling loop that regulated the macrophage response. Removing this signaling loop enhanced the pro-inflammatory response to LPS. Finally, we found that this DAT-immune axis was disrupted in disease. Collectively, our data introduce a novel role for DAT in the regulation of innate immunity during health and disease.

scholarly journals Characterization of Novel RFamide Peptides in the Central Nervous System of the Brown Hagfish: Isolation, Localization, and Functional Analysis

Endocrinology ◽  
2011 ◽  
Vol 152 (11) ◽  
pp. 4252-4264 ◽  
Author(s):  
Tomohiro Osugi ◽  
Katsuhisa Uchida ◽  
Masumi Nozaki ◽  
Kazuyoshi Tsutsui
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Brain Regions ◽  
Peptide Group ◽  
Neuropeptide Ff ◽  
Peptide Precursor ◽  
Molecular Phylogenetic ◽  
The Central Nervous System

RFamide (RFa) peptides play various important roles in the central nervous system in both invertebrates and vertebrates. However, there is no evidence of the existence of any RFamide peptide in the brain of hagfish, one of the oldest lineages of vertebrates. In this study, we sought to identify novel RFamide peptides from the brains of hagfish (Paramyxine atami). We identified four novel RFamide peptides, which had the C-terminal Pro-Gln-Arg-Phe-NH2 structure. cDNA cloning revealed that the identified RFamide peptides are encoded in two types of cDNA. Molecular phylogenetic analysis of the two precursors indicated that the hagfish RFamide peptides belong to the PQRFamide peptide group that includes mammalian neuropeptide FF and AF. Based on immunohistochemistry and in situ hybridization, hagfish PQRFamide peptide precursor mRNA and its translated peptides were localized in the infundibular nucleus of the hypothalamus. Immunoreactive fibers were terminated on blood vessels in the infundibular nucleus. Dense immunoreactive fibers were also observed in other brain regions. We further showed that one of the hagfish PQRFamide peptides significantly stimulated the expression of gonadotropin-β mRNA in the cultured hagfish pituitary. These results indicate that the control mechanism of gonadotropin expression by a hypothalamic neuropeptide evolved in the agnathan brain. This is the first evidence describing the identification of RFamide peptides in the hagfish brain. This is also the first report showing the regulation of gonadotropin expression by a homolog of neuropeptide FF that belongs to the PQRFamide peptide group in any vertebrate.

Characterization of the symbiontRickettsiain the mirid bugNesidiocoris tenuis(Reuter) (Heteroptera: Miridae)

2014 ◽  
Vol 104 (6) ◽  
pp. 681-688 ◽  
Author(s):  
A. Caspi-Fluger ◽  
M. Inbar ◽  
S. Steinberg ◽  
Y. Friedmann ◽  
M. Freund ◽  
...  
Keyword(s):  
Biological Control ◽  
Developmental Stages ◽  
Bacterial Symbionts ◽  
Nesidiocoris Tenuis ◽  
Augmentative Release ◽  
Polymerase Chain ◽  
Bacterial Genes ◽  
Rickettsia Bellii

AbstractNesidiocoris tenuis(Reuter) (Heteroptera: Miridae) is an omnivorous insect used for biological control. Augmentative release and conservation ofN. tenuishave been used for pest control in tomato crops. Intracellular bacterial symbionts of arthropods are common in nature and have diverse effects on their hosts; in some cases they can dramatically affect biological control. Fingerprinting methods showed that the symbiotic complex associated withN. tenuisincludesWolbachiaandRickettsia. RickettsiaofN. tenuiswas further characterized by sequencing the16S rRNAandgltAbacterial genes, measuring its amount in different developmental stages of the insect by real-time polymerase chain reaction, and localizing the bacteria in the insect's body by fluorescencein situhybridization. TheRickettsiainN. tenuisexhibited 99 and 96% similarity of both sequenced genes toRickettsia belliiandRickettsiareported fromBemisia tabaci, respectively. The highest amount ofRickettsiawas measured in the 5th instar and adult, and the symbionts could be detected in the host gut and ovaries. Although the role played byRickettsiain the biology ofN. tenuisis currently unknown, their high amount in the adults and localization in the gut suggest that they may have a nutritional role in this insect.

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