scholarly journals Identification and characterisation of the cryptic Golgi Apparatus in Naegleria gruberi

2017 ◽  
Author(s):  
Emily K. Herman ◽  
Lyto Yiangou ◽  
Diego M. Cantoni ◽  
Christopher N. Miller ◽  
Francine Marciano-Cabral ◽  
...  
Keyword(s):  
Golgi Apparatus ◽  
Membrane Trafficking ◽  
Brefeldin A ◽  
Sampling Point ◽  
Eukaryotic Lineage ◽  
Separate Species ◽  
Immuno Electron Microscopy ◽  
First Time ◽  
Naegleria Gruberi ◽  
Tubular Form

AbstractAlthough the Golgi apparatus has a conserved morphology of flattened stacked cisternae in the vast majority of eukaryotes, the organelle has lost the stacked organization in several eukaryotic lineages raising the question of what range of morphologies is possible for the Golgi. In order to understand this range of organellar diversity, it is necessary to characterise the Golgi in many different lineages. Here we identify the Golgi apparatus in Naegleria, the first description of an unstacked Golgi organelle in a non-parasitic eukaryote, other than fungi. We provide a comprehensive list of Golgi-associated membrane trafficking genes encoded in two separate species of Naegleria and transcriptomic support to show that nearly all are expressed in mouse-passaged N. fowleri cells. We then study distribution of the Golgi marker NgCOPB by fluorescence, identifying membranous structures that can be disrupted by Brefeldin A treatment consistent with Golgi localisation. Confocal and immuno-electron microscopy revealed that NgCOPB is localized to membranous structures consistent with tubules. Our data not only identify the Golgi organelle for the first time in this major eukaryotic lineage, but also provide the rare example of a tubular form of the organelle representing an important sampling point for the comparative understanding of Golgi organellar diversity.

scholarly journals Maintenance of Golgi structure and function depends on the integrity of ER export

2001 ◽  
Vol 155 (4) ◽  
pp. 557-570 ◽  
Author(s):  
Theresa H. Ward ◽  
Roman S. Polishchuk ◽  
Steve Caplan ◽  
Koret Hirschberg ◽  
Jennifer Lippincott-Schwartz
Keyword(s):  
Steady State ◽  
Golgi Apparatus ◽  
Membrane Trafficking ◽  
Imaging Techniques ◽  
Brefeldin A ◽  
Er Exit Sites ◽  
Selective Membrane ◽  
Golgi Structure ◽  
And Function ◽  
Er Export

The Golgi apparatus comprises an enormous array of components that generate its unique architecture and function within cells. Here, we use quantitative fluorescence imaging techniques and ultrastructural analysis to address whether the Golgi apparatus is a steady-state or a stable organelle. We found that all classes of Golgi components are dynamically associated with this organelle, contrary to the prediction of the stable organelle model. Enzymes and recycling components are continuously exiting and reentering the Golgi apparatus by membrane trafficking pathways to and from the ER, whereas Golgi matrix proteins and coatomer undergo constant, rapid exchange between membrane and cytoplasm. When ER to Golgi transport is inhibited without disrupting COPII-dependent ER export machinery (by brefeldin A treatment or expression of Arf1[T31N]), the Golgi structure disassembles, leaving no residual Golgi membranes. Rather, all Golgi components redistribute into the ER, the cytoplasm, or to ER exit sites still active for recruitment of selective membrane-bound and peripherally associated cargos. A similar phenomenon is induced by the constitutively active Sar1[H79G] mutant, which has the additional effect of causing COPII-associated membranes to cluster to a juxtanuclear region. In cells expressing Sar1[T39N], a constitutively inactive form of Sar1 that completely disrupts ER exit sites, Golgi glycosylation enzymes, matrix, and itinerant proteins all redistribute to the ER. These results argue against the hypothesis that the Golgi apparatus contains stable components that can serve as a template for its biogenesis. Instead, they suggest that the Golgi complex is a dynamic, steady-state system, whose membranes can be nucleated and are maintained by the activities of the Sar1–COPII and Arf1–coatomer systems.

Morphological and molecular characteristics of Bastiania sinensis sp. n., Tripyla aquatica Brzeski & Winiszewska-Ślipińska, 1993 and T. setifera Bütschli, 1873 (Nematoda: Triplonchida) from Shanxi Province, North China

Nematology ◽  
2021 ◽  
pp. 1-23
Author(s):  
Mei Na Liu ◽  
Yu Mei Xu ◽  
Zeng Qi Zhao ◽  
Jian Ming Wang
Keyword(s):  
North China ◽  
Large Subunit ◽  
Small Subunit ◽  
Rrna Genes ◽  
Shanxi Province ◽  
Molecular Characteristics ◽  
Separate Species ◽  
Using Data ◽  
First Time ◽  
Respective Species

Summary This paper describes a new species of Bastiania, presents a new record and redescribes a known species of Tripyla. These nematodes are all in the order Triplonchida and were collected from Shanxi Province, North China. Bastiania sinensis sp. n. is characterised by having the female with a relatively slender body 1049-1295 μm long, dorsally arcuate after heat relaxation, with outer labial setae and cephalic setae in a single circle, an oval amphid, 7-8 laterodorsal cervical setae scattered in the pharyngeal region, orthometamenes and pseudocoelomocytes present, tail conoid with a mucron 1-2 μm long, two pairs of caudal setae present, a = 58.1-75.5, b = 4.0-4.6, c = 12.7-19.7, c′ = 4.1-7.8 and V = 61.1-67.7. Males were not found. Tripyla aquatica is recorded for the first time from China, and is redescribed. Tripyla setifera has been reported from China but without a detailed description – now provided. In addition, phylogenetic relationships among the species were analysed using data from the near full length small subunit (SSU) and D2-D3 segments of large subunit (LSU) of rRNA genes. Bastiania sinensis sp. n. is monophyletic with the Bastiania sequences available in GenBank, but is on an independent branch supporting its status as a separate species; T. aquatica and T. setifera are monophyletically clustered with known Tripyla species and grouped together with sequences from their respective species.

Important role for the V-type H+-ATPase and the Golgi apparatus in the recycling of PTH/PTHrP receptor

2004 ◽  
Vol 286 (5) ◽  
pp. E704-E710 ◽  
Author(s):  
Hesham A. W. Tawfeek ◽  
Abdul B. Abou-Samra
Keyword(s):  
Golgi Apparatus ◽  
Fluorescent Protein ◽  
Brefeldin A ◽  
Receptor Recycling ◽  
Bafilomycin A1 ◽  
Coated Pits ◽  
Concanamycin A ◽  
Hydrogen Atpase ◽  
Green Fluorescent ◽  
Pthrp Receptor

Our previous studies demonstrated that a green fluorescent protein-tagged parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor stably expressed in LLCPK-1 cells undergoes agonist-dependent internalization into clathrin-coated pits. The subcellular localization of the internalized PTH/PTHrP receptor is not known. In the present study, we explored the intracellular pathways of the internalized PTH/PTHrP receptor. Using immunofluorescence and confocal microscopy, we show that the internalized receptors localize at a juxtanuclear compartment identified as the Golgi apparatus. The receptors do not colocalize with lysosomes. Furthermore, whereas the internalized receptors exhibit rapid recycling, treatment with proton pump inhibitors (bafilomycin-A1 and concanamycin A) or brefeldin A, Golgi disrupting agents, reduces PTH/PTHrP receptor recycling. Together, these data indicate an important role for the vacuolar-type hydrogen-ATPase and the Golgi apparatus in postendocytic PTH/PTHrP receptor recovery.

scholarly journals An AP-3-dependent mechanism drives synaptic-like microvesicle biogenesis in pancreatic islet β-cells

2010 ◽  
Vol 299 (1) ◽  
pp. E23-E32 ◽  
Author(s):  
Arthur T. Suckow ◽  
Branch Craige ◽  
Victor Faundez ◽  
William J. Cain ◽  
Steven D. Chessler
Keyword(s):  
Synaptic Vesicle ◽  
Pancreatic Islet ◽  
Membrane Trafficking ◽  
Protein Complexes ◽  
Brefeldin A ◽  
Adaptor Protein ◽  
Β Cells ◽  
Β Cell ◽  
Subunit Expression ◽  
Bona Fide

Pancreatic islet β-cells contain synaptic-like microvesicles (SLMVs). The origin, trafficking, and role of these SLMVs are poorly understood. In neurons, synaptic vesicle (SV) biogenesis is mediated by two different cytosolic adaptor protein complexes, a ubiquitous AP-2 complex and the neuron-specific AP-3B complex. Mice lacking AP-3B subunits exhibit impaired GABAergic (inhibitory) neurotransmission and reduced neuronal vesicular GABA transporter (VGAT) content. Since β-cell maturation and exocytotic function seem to parallel that of the inhibitory synapse, we predicted that AP-3B-associated vesicles would be present in β-cells. Here, we test the hypothesis that AP-3B is expressed in islets and mediates β-cell SLMV biogenesis. A secondary aim was to test whether the sedimentation properties of INS-1 β-cell microvesicles are identical to those of bona fide SLMVs isolated from PC12 cells. Our results show that the two neuron-specific AP-3 subunits β3B and μ3B are expressed in β-cells, the first time these proteins have been found to be expressed outside the nervous system. We found that β-cell SLMVs share the same sedimentation properties as PC12 SLMVs and contain SV proteins that sort specifically to AP-3B-associated vesicles in the brain. Brefeldin A, a drug that interferes with AP-3-mediated SV biogenesis, inhibits the delivery of AP-3 cargoes to β-cell SLMVs. Consistent with a role for AP-3 in the biogenesis of GABAergic SLMV in β-cells, INS-1 cell VGAT content decreases upon inhibition of AP-3 δ-subunit expression. Our findings suggest that β-cells and neurons share molecules and mechanisms important for mediating the neuron-specific membrane trafficking pathways that underlie synaptic vesicle formation.

scholarly journals Brefeldin A inhibits transport of the glycophorin-binding protein from Plasmodium falciparum into the host erythrocyte

1994 ◽  
Vol 300 (3) ◽  
pp. 821-826 ◽  
Author(s):  
J Benting ◽  
D Mattei ◽  
K Lingelbach
Keyword(s):  
Plasmodium Falciparum ◽  
Golgi Apparatus ◽  
Host Cell ◽  
Protein Secretion ◽  
Secretory Pathway ◽  
Binding Protein ◽  
Brefeldin A ◽  
Cell Cytoplasm ◽  
Parasite Cell ◽  
Host Cell Cytoplasm

Plasmodium falciparum, a protozoan parasite of the human erythrocyte, causes the most severe form of malaria. During its intraerythrocytic development, the parasite synthesizes proteins which are exported into the host cell. The compartments involved in the secretory pathway of P. falciparum are still poorly characterized. A Golgi apparatus has not been identified, owing to the lack of specific protein markers and Golgi-specific post-translational modifications in the parasite. The fungal metabolite brefeldin A (BFA) is known to inhibit protein secretion in higher eukaryotes by disrupting the integrity of the Golgi apparatus. We have used the parasite-encoded glycophorin-binding protein (GBP), a soluble protein found in the host cell cytoplasm, as a marker to investigate the effects of BFA on protein secretion in the intracellular parasite. In the presence of BFA, GBP was not transported into the erythrocyte, but remained inside the parasite cell. The effect caused by BFA was reversible, and the protein could be chased into the host cell cytoplasm within 30 min. Transport of GBP from the BFA-sensitive site into the host cell did not require protein synthesis. Similar observations were made when infected erythrocytes were incubated at 15 degrees C. Incubation at 20 degrees C resulted in a reduction rather than a complete block of protein export. The relevance of our findings to the identification of compartments involved in protein secretion from the parasite cell is discussed.

scholarly journals Fauna of gamasina mites (Gamasoidea) of small mammals of a natural region «Samarskaya Luka»

2016 ◽  
Vol 5 (2) ◽  
pp. 57-63
Author(s):  
Irina Anatol’evna Sorokopud
Keyword(s):  
Small Mammals ◽  
Water Vole ◽  
Separate Species ◽  
Natural Region ◽  
Mole Vole ◽  
Ecological Complex ◽  
Tundra Vole ◽  
First Time ◽  
Samarskaya Luka

Data on fauna of gamasina mites parasitizing on small mammals are provided in article. Data are obtained during long-term monitoring of an ecological complex of small mammals of a natural and territorial complex Samarskaya Luka, which is realized since 1999. Also data on fauna of gamasina mites in 14 years of researches (2000-2003, 2005, 2007-2015 years), including literary data about the gamasina mites of this territory, are generalized and analysed. Specific accessory of 7439 individuals of mites, seven childbirth of three families relating to fifteen types is defined. Parasitizing the gamasina mites on 16 species of small mammals, including groups rodents, soricomorpha, carnivora (least weasel) is established. Domination of separate species of mites is revealed, so to eudominant Laelaps hilaris and Hirstionyssus isabellinus are belong, dominants are - Hirstionyssus ellobii , Haemolaelaps glasgowi , Hyperlaelaps arvalis , L. agilis . Myonyssus rossicus , L. algericus , Haemogamasus ambulans and Hirstionyssus macedonicus were extremely rare species met in single copies. For the first time for this territory types of H. ellobii , L. micromydis , H. ambulans , H. macedonicus are noted. Conclusions on degree of a confinedness of parasites to certain types of hosts are drawn. Monohostal types: L. agilis (the host - a yellow-necked mouse), L. muris (the host - a tundra vole and a water vole), L. micromydis (the host - a small mouse), L. algericus (the host - a house mouse), H. arvalis (the host - a tundra vole and a water vole), H. ellobii (the host - a northern mole vole). Polihostal: H. musculi , L. hilaris , H. isabellinus , H. glasgowi , E. stabularis , H. nidi .

scholarly journals Lava Lamp, a Novel Peripheral Golgi Protein, Is Required for Drosophila melanogaster Cellularization

2000 ◽  
Vol 151 (4) ◽  
pp. 905-918 ◽  
Author(s):  
John C. Sisson ◽  
Christine Field ◽  
Richard Ventura ◽  
Anne Royou ◽  
William Sullivan
Keyword(s):  
Membrane Trafficking ◽  
Biochemical Analysis ◽  
Potent Inhibitor ◽  
Animal Cell ◽  
Coiled Coil ◽  
Brefeldin A ◽  
Microtubule Associated Proteins ◽  
Golgi Bodies ◽  
Associated Proteins ◽  
Golgi Protein

Drosophila cellularization and animal cell cytokinesis rely on the coordinated functions of the microfilament and microtubule cytoskeletal systems. To identify new proteins involved in cellularization and cytokinesis, we have conducted a biochemical screen for microfilament/microtubule-associated proteins (MMAPs). 17 MMAPs were identified; seven have been previously implicated in cellularization and/or cytokinesis, including KLP3A, Anillin, Septins, and Dynamin. We now show that a novel MMAP, Lava Lamp (Lva), is also required for cellularization. Lva is a coiled-coil protein and, unlike other proteins previously implicated in cellularization or cytokinesis, it is Golgi associated. Our functional analysis shows that cellularization is dramatically inhibited upon injecting anti–Lva antibodies (IgG and Fab) into embryos. In addition, we show that brefeldin A, a potent inhibitor of membrane trafficking, also inhibits cellularization. Biochemical analysis demonstrates that Lva physically interacts with the MMAPs Spectrin and CLIP190. We suggest that Lva and Spectrin may form a Golgi-based scaffold that mediates the interaction of Golgi bodies with microtubules and facilitates Golgi-derived membrane secretion required for the formation of furrows during cellularization. Our results are consistent with the idea that animal cell cytokinesis depends on both actomyosin-based contraction and Golgi-derived membrane secretion.

Sign in / Sign up

Export Citation Format

Share Document