scholarly journals AP2 hemicomplexes contribute independently to synaptic vesicle endocytosis

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
Mingyu Gu ◽  
Qiang Liu ◽  
Shigeki Watanabe ◽  
Lin Sun ◽  
Gunther Hollopeter ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Synaptic Vesicle ◽  
Synaptic Vesicles ◽  
Nematode Caenorhabditis Elegans ◽  
Α Subunit ◽  
Clathrin Adaptor ◽  
Simultaneous Loss

The clathrin adaptor complex AP2 is thought to be an obligate heterotetramer. We identify null mutations in the α subunit of AP2 in the nematode Caenorhabditis elegans. α-adaptin mutants are viable and the remaining μ2/β hemicomplex retains some function. Conversely, in μ2 mutants, the alpha/sigma2 hemicomplex is localized and is partially functional. α-μ2 double mutants disrupt both halves of the complex and are lethal. The lethality can be rescued by expression of AP2 components in the skin, which allowed us to evaluate the requirement for AP2 subunits at synapses. Mutations in either α or μ2 subunits alone reduce the number of synaptic vesicles by about 30%; however, simultaneous loss of both α and μ2 subunits leads to a 70% reduction in synaptic vesicles and the presence of large vacuoles. These data suggest that AP2 may function as two partially independent hemicomplexes.

scholarly journals μ2 adaptin facilitates but is not essential for synaptic vesicle recycling in Caenorhabditis elegans

2008 ◽  
Vol 183 (5) ◽  
pp. 881-892 ◽  
Author(s):  
Mingyu Gu ◽  
Kim Schuske ◽  
Shigeki Watanabe ◽  
Qiang Liu ◽  
Paul Baum ◽  
...  
Keyword(s):  
Nervous System ◽  
Caenorhabditis Elegans ◽  
Synaptic Vesicle ◽  
Synaptic Vesicles ◽  
Evoked Responses ◽  
Nematode Caenorhabditis Elegans ◽  
Synaptic Vesicle Recycling ◽  
Specific Loss ◽  
Vesicle Recycling ◽  
Cargo Proteins

Synaptic vesicles must be recycled to sustain neurotransmission, in large part via clathrin-mediated endocytosis. Clathrin is recruited to endocytic sites on the plasma membrane by the AP2 adaptor complex. The medium subunit (μ2) of AP2 binds to cargo proteins and phosphatidylinositol-4,5-bisphosphate on the cell surface. Here, we characterize the apm-2 gene (also called dpy-23), which encodes the only μ2 subunit in the nematode Caenorhabditis elegans. APM-2 is highly expressed in the nervous system and is localized to synapses; yet specific loss of APM-2 in neurons does not affect locomotion. In apm-2 mutants, clathrin is mislocalized at synapses, and synaptic vesicle numbers and evoked responses are reduced to 60 and 65%, respectively. Collectively, these data suggest AP2 μ2 facilitates but is not essential for synaptic vesicle recycling.

scholarly journals UNC-11, a Caenorhabditis elegans AP180 Homologue, Regulates the Size and Protein Composition of Synaptic Vesicles

1999 ◽  
Vol 10 (7) ◽  
pp. 2343-2360 ◽  
Author(s):  
Michael L. Nonet ◽  
Andrea M. Holgado ◽  
Faraha Brewer ◽  
Craig J. Serpe ◽  
Betty A. Norbeck ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Synaptic Vesicle ◽  
Neurotransmitter Release ◽  
Synaptic Vesicles ◽  
Adaptor Protein ◽  
Protein Composition ◽  
Mammalian Brain ◽  
Snare Protein ◽  
Vesicle Biogenesis ◽  
Clathrin Adaptor

The unc-11 gene of Caenorhabditis elegans encodes multiple isoforms of a protein homologous to the mammalian brain-specific clathrin-adaptor protein AP180. The UNC-11 protein is expressed at high levels in the nervous system and at lower levels in other tissues. In neurons, UNC-11 is enriched at presynaptic terminals but is also present in cell bodies. unc-11mutants are defective in two aspects of synaptic vesicle biogenesis. First, the SNARE protein synaptobrevin is mislocalized, no longer being exclusively localized to synaptic vesicles. The reduction of synaptobrevin at synaptic vesicles is the probable cause of the reduced neurotransmitter release observed in these mutants. Second,unc-11 mutants accumulate large vesicles at synapses. We propose that the UNC-11 protein mediates two functions during synaptic vesicle biogenesis: it recruits synaptobrevin to synaptic vesicle membranes and it regulates the size of the budded vesicle during clathrin coat assembly.

On the nature of undead cells in the nematode Caenorhabditis elegans

1991 ◽  
Vol 331 (1261) ◽  
pp. 263-271 ◽  
Keyword(s):  
Cell Death ◽  
Caenorhabditis Elegans ◽  
Epithelial Cell ◽  
Programmed Cell Death ◽  
Synaptic Vesicles ◽  
Loss Of Function ◽  
Cell Type ◽  
Nematode Caenorhabditis Elegans ◽  
Characteristic Features ◽  
Morphological Differences

During the course of normal embryonic and post-embryonic development, 131 cells in a Caenorhabditis elegans hermaphrodite undergo programmed cell death. Loss of function mutations in either of the genes ced-3 or ced-4 abolish cell deaths, enabling these ‘undead’ cells to survive and be incorporated into the adult with no obvious deleterious consequences. Ultrastructural reconstructions have shown that undead cells exhibit many differentiated characteristics. Most of the reconstructed cells appeared to be neurons with all the characteristic features associated with such cells, such as processes, synaptic vesicles and presynaptic specializations. However, clear morphological differences were seen among the undead neurons, suggesting a diversity of cell type. One of the reconstructed cells was a rectal epithelial cell, which had displaced its lineal sister that normally functions in this role. Removal of the ability to undergo programmed cell death by mutation therefore reveals a diversity of cryptic differentiated states that are acquired by cells that normally are destined to die.

Polyunsaturated fatty acids and neurotransmission in Caenorhabditis elegans

2006 ◽  
Vol 34 (1) ◽  
pp. 77-80 ◽  
Author(s):  
E. Marza ◽  
G.M. Lesa
Keyword(s):  
Fatty Acids ◽  
Mental Retardation ◽  
Cognitive Impairment ◽  
Fatty Acid ◽  
Caenorhabditis Elegans ◽  
Synaptic Vesicles ◽  
Acid Metabolism ◽  
Neuronal Dysfunction ◽  
Nematode Caenorhabditis Elegans ◽  
Low Levels

Changes in PUFA (polyunsaturated fatty acid) metabolism can cause mental retardation and cognitive impairment. However, it is still unclear why altered levels of PUFAs result in neuronal dysfunction. Recent studies on the nematode Caenorhabditis elegans suggest that PUFA depletion may cause cognitive impairment by compromising communication among neurons. Pharmacological and electrophysiological experiments showed that animals devoid of most PUFAs release abnormally low levels of neurotransmitters. In addition, ultrastructural analysis revealed that synapses in these mutants are severely depleted of synaptic vesicles. The conclusion of these studies is that PUFAs are required to maintain a normal pool of synaptic vesicles at pre-synaptic sites, thus ensuring efficient neurotransmission.

scholarly journals Mutations in Synaptojanin Disrupt Synaptic Vesicle Recycling

2000 ◽  
Vol 150 (3) ◽  
pp. 589-600 ◽  
Author(s):  
Todd W. Harris ◽  
Erika Hartwieg ◽  
H. Robert Horvitz ◽  
Erik M. Jorgensen
Keyword(s):  
Plasma Membrane ◽  
Caenorhabditis Elegans ◽  
Synaptic Vesicle ◽  
Synaptic Vesicles ◽  
Vesicle Trafficking ◽  
Synaptic Vesicle Recycling ◽  
Vesicle Recycling ◽  
Synaptojanin 1

Synaptojanin is a polyphosphoinositide phosphatase that is found at synapses and binds to proteins implicated in endocytosis. For these reasons, it has been proposed that synaptojanin is involved in the recycling of synaptic vesicles. Here, we demonstrate that the unc-26 gene encodes the Caenorhabditis elegans ortholog of synaptojanin. unc-26 mutants exhibit defects in vesicle trafficking in several tissues, but most defects are found at synaptic termini. Specifically, we observed defects in the budding of synaptic vesicles from the plasma membrane, in the uncoating of vesicles after fission, in the recovery of vesicles from endosomes, and in the tethering of vesicles to the cytoskeleton. Thus, these results confirm studies of the mouse synaptojanin 1 mutants, which exhibit defects in the uncoating of synaptic vesicles (Cremona, O., G. Di Paolo, M.R. Wenk, A. Luthi, W.T. Kim, K. Takei, L. Daniell, Y. Nemoto, S.B. Shears, R.A. Flavell, D.A. McCormick, and P. De Camilli. 1999. Cell. 99:179–188), and further demonstrate that synaptojanin facilitates multiple steps of synaptic vesicle recycling.

Interaction of a Free-Living Soil Nematode, Caenorhabditis elegans, with Surrogates of Foodborne Pathogenic Bacteria

2003 ◽  
Vol 66 (9) ◽  
pp. 1543-1549 ◽  
Author(s):  
GARY L. ANDERSON ◽  
KRISHAUN N. CALDWELL ◽  
LARRY R. BEUCHAT ◽  
PHILLIP L. WILLIAMS
Keyword(s):  
Caenorhabditis Elegans ◽  
Young Adult ◽  
Salmonella Typhimurium ◽  
Avirulent Strain ◽  
Soil Nematode ◽  
Gram Negative Bacteria ◽  
Nematode Caenorhabditis Elegans ◽  
Free Living ◽  
Gram Positive ◽  
Gram Negative

Free-living nematodes may harbor, protect, and disperse bacteria, including those ingested and passed in viable form in feces. These nematodes are potential vectors for human pathogens and may play a role in foodborne diseases associated with fruits and vegetables eaten raw. In this study, we evaluated the associations between a free-living soil nematode, Caenorhabditis elegans, and Escherichia coli, an avirulent strain of Salmonella Typhimurium, Listeria welshimeri, and Bacillus cereus. On an agar medium, young adult worms quickly moved toward colonies of all four bacteria; over 90% of 3-day-old adult worms entered colonies within 16 min after inoculation. After 48 h, worms moved in and out of colonies of L. welshimeri and B. cereus but remained associated with E. coli and Salmonella Typhimurium colonies for at least 96 h. Young adult worms fed on cells of the four bacteria suspended in K medium. Worms survived and reproduced with the use of nutrients derived from all test bacteria, as determined for eggs laid by second-generation worms after culturing for 96 h. Development was slightly slower for worms fed gram-positive bacteria than for worms fed gram-negative bacteria. Worms that fed for 24 h on bacterial lawns formed on tryptic soy agar dispersed bacteria over a 3-h period when they were transferred to a bacteria-free agar surface. The results of this study suggest that C. elegans and perhaps other free-living nematodes are potential vectors for both gram-positive and gram-negative bacteria, including foodborne pathogens in soil.

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