-
[
Worm Breeder's Gazette,
1993]
ceh 6 expression and generation of a knock-out Thomas Burglin, Ron Plasterk*, Gary Ruvkun Dept. of Molecular Biology, Wellman 8, Massachusetts General Hospital, Boston, MA, 02114, USA, and *Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, The Netherlands.
-
[
The New York Times,
1997]
His tall figure bent over a computer screen in his laboratory at the Massachusetts General Hospital, Dr. Gary Ruvkun rummages through a distant genetic data base for matches to a gene he believes is involved in diabetes. ?You learn how to read these as they are ratcheting by,? he says, while lines of data streak up his screen. ?I think MTV is good training.?
-
[
C.elegans Neuronal Development Meeting,
2008]
The ability to record neuronal activity and behavior simultaneously is one of the few remaining obstacles to the goal of a comprehensive understanding of the C. elegans nervous system. Such recordings have not so far been achieved at high resolution (63x) in unrestrained, freely crawling nematodes, because the target neuron moves through the field of view in a fraction of a second. Here we present a new tracking system, developed in co-operation with Applied Scientific Instruments (Eugene, OR), that solves this problem by recentering the neuronal image in real time. The system operates by diverting a fraction of the light that usually goes to the Ca imaging camera to a quadrant photomultiplier tube (PMT) which functions as a high-speed four-pixel camera. The differences in light intensity among the four quadrants are used to monitor the displacement of the target with respect to the optical axis of the microscope. The displacement signal is then converted into commands to a motorized stage which compensate for the displacement in real time. Because the system is realized in analog circuitry, and requires no image processing, recentering of the target is extremely fast, occurring on the timescale of milliseconds. A second camera records the worm''s behavior via a wide-field objective. The two cameras are synchronized to provide a continuous record of neuronal activity and behavior. The system is augmented by image registration software that corrects for image rotation associated with head swings and other correlates of locomotion to keep cells of interest fixed for the duration of the movie. As proof of principle, we have successfully recorded Ca transients in interneurons with cell bodies in the head, and motor neurons with cell bodies in the ventral cord. The new system is likely to accelerate studies of the neuronal control of behavior in C. elegans. NIH MH51383.
-
[
Worm Breeder's Gazette,
1991]
Ed Hedgecock came to Tokyo in January for the 2nd seminar on Molecular and Developmental Neurobiology and his cell migration work impressed scientists of other animal field. Sydney Brenner won the Kyoto Prize and came to Japan on 23 of October for receiving the Prize on his fourth visit in this year. He talked with his excellent joke about telescope in Astronomy and microscope in Biology. Sydney is the man who gave the main lecture in the First Meeting of Japan Molecular Biology Society. Iva Greenwald came to Japan for attending the Naito Foundation International Workshop on Morphogenesis Program in early November together with her fiance Gary Struhl. It is true that the relation between Drosophila and Caenorhabditis is very good in U.S. and Japan. Some nematode scientists in Japan got grant from Drosophila project. John Sulston gave the lecture entitled 'The Genome of Caenorhabditis' on 28th of November at the main invited lecture of the Thirteenth Meeting of Japan Molecular Biology Society in Kyoto International Congress Hall. The lecture impressed many Japanese scientists especially his 'The Logical Next Step: Genome Sequencing'. On the 27th night, 34 worm people assembled to the worm party for welcoming to him. Sixteen papers from five labs were presented at the meeting.
-
[
RNA Biol,
2019]
Worm biologists from the United States, Canada, and the United Kingdom gathered at the Colorado State University Todos Santos Center in Baja California Sur, Mexico, April 3-5, 2019 for the Todos Santos Small RNA Symposium. Meeting participants, many of whom were still recovering from the bomb cyclone that struck a large swath of North America just days earlier, were greeted by the warmth and sunshine that is nearly ubiquitous in the sleepy seaside town of Todos Santos. With only 24 speakers, the meeting had the sort of laid-back vibe you might expect amongst the palm trees and ocean breeze of the Pacific coast of Mexico. The meeting started with tracing the laboratory lineages of participants. Not surprisingly, the most common parental lineages represented at the meeting were Dr. Craig Mello, Dr. Gary Ruvkun, and Dr. Victor Ambros, whom, together with Dr. Andy Fire and Dr. David Baulcombe, pioneered the small RNA field. In sad irony, on the closing day of the meeting, participants were met with the news of Dr. Sydney Brenner's passing. By establishing the worm, <i>Caenorhabditis elegans</i>, as a model system Dr. Brenner paved the way for much of the research discussed here.
-
Berriman, Matt, Howe, Kevin, Kersey, Paul, Stein, Lincoln, Harris, Todd, Sternberg, Paul, Schedl, Tim
[
International Worm Meeting,
2015]
WormBase has existed for 15 years and has evolved in many ways. The new website is fully operational and has made the process of adding new data types, displays, and tools easier. Behind the scenes we are piloting an overhaul of the underlying database infrastructure to allow us to handle the ever increasing data, have the website perform faster, and allow more frequent updates of information. This is a critical time for the project, as we face considerable pressure from two directions. The first is that our funders really want us to do more with less. We are responding to this by leading the way in making curation (the process of extracting information from papers and data sets into computable form) more efficient using a new version of Textpresso (to be released later this calendar year); by discussing with other model organism information resources ways to work together to be more efficient and inter-connected; and by seeking additional sources of funding. The second, delightful, pressure is an increase in data and results generated by the C. elegans and nematode communities. While we are handling this increase by changes in our software for curation, the database infrastructure, and the website, we do need your help. Many of you have helped us over the last few years to identify data in your papers or by sending us data directly. We now need you to help with a few types of information by submitting the data via specially designed, user-friendly forms that ensure good quality and the use of standard terminology. In particular, we have a large backlog of uncurated information associating alleles with phenotypes. We pledge to make this process as painless as possible, and to improve WormBase's description of phenotypes with your feedback, starting at this meeting at the WormBase booth, workshops and posters. With your help, continual improvement of our efficiency, and additional sources of funding, we are optimistic that we can do much more with even somewhat less effort.Consortium: Paul Davis, Michael Paulini, Gary Williams, Bruce Bolt, Thomas Down, Jane Lomax, Todd Harris, Sibyl Gao, Scott Cain, Xiaodong Wang, Karen Yook, Juancarlos Chan, Wen Chen, Chris Grove, Mary Ann Tuli, Kimberly Van Auken, D. Wang, Ranjana Kishore, Raymond Lee, John DeModena, James Done, Yuling Li, H.-M. Mueller, Cecilia Nakamura, Daniela Raciti, Gary Schindelman.
-
[
Front Cell Dev Biol,
2022]
Axon-dendrite formation is a crucial milestone in the life history of neurons. During this process, historically referred as "the establishment of polarity," newborn neurons undergo biochemical, morphological and functional transformations to generate the axonal and dendritic domains, which are the basis of neuronal wiring and connectivity. Since the implementation of primary cultures of rat hippocampal neurons by Gary Banker and Max Cowan in 1977, the community of neurobiologists has made significant achievements in decoding signals that trigger axo-dendritic specification. External and internal cues able to switch on/off signaling pathways controlling gene expression, protein stability, the assembly of the polarity complex (i.e., PAR3-PAR6-aPKC), cytoskeleton remodeling and vesicle trafficking contribute to shape the morphology of neurons. Currently, the culture of hippocampal neurons coexists with alternative model systems to study neuronal polarization in several species, from single-cell to whole-organisms. For instance, <i>in vivo</i> approaches using <i>C. elegans</i> and <i>D. melanogaster,</i> as well as <i>in situ</i> imaging in rodents, have refined our knowledge by incorporating new variables in the polarity equation, such as the influence of the tissue, glia-neuron interactions and three-dimensional development. Nowadays, we have the unique opportunity of studying neurons differentiated from human induced pluripotent stem cells (hiPSCs), and test hypotheses previously originated in small animals and propose new ones perhaps specific for humans. Thus, this article will attempt to review critical mechanisms controlling polarization compiled over decades, highlighting points to be considered in new experimental systems, such as hiPSC neurons and human brain organoids.
-
[
International C. elegans Meeting,
1995]
We are studying the
nhr-1 (nuclear hormone receptor) gene, one of a number of steroid receptor homologues identified in C. elegans. Sequencing of the cDNA has shown interesting features for this putative "orphan" receptor, including an unusual sequence in the P box of the DNA binding domain, which might indicate a very different DNA binding specificity and function, and a very long 3' untranslated region. The
nhr-1 gene has been mapped to the right arm of the X chromosome, very close to
sup-10. We are now characterizing the genomic organization of
nhr-1 in C. elegans. We have also initiated work with C. briggsae in order to identify conserved, functionally important regions. The presence of an apparent homologue in C. briggsae was of special interest to us, given the unusual properties of
nhr-1 (different P box, the long 3'UT region), and the apparent viability of homozygous deficiencies in this region around
sup-10 (C. Cummins and P. Anderson). The C. elegans
nhr-1 gene appears to contain 12 exons spread over 9.5 kb, including several relatively large introns, 1.3 to 2.3 kb in size. Work to date with genomic clones from C. briggsae shows conserved exons which share approximately 81-84% homology at the nucleotide level with the
nhr-1 gene in C. elegans. This conservation between species, in the absence of a lethal phenotype for the homozygous deficiency, suggests that the
nhr-1 gene may nevertheless have an evolutionarily relevant function. We wish to thank Ann Sluder, Gary Ruvkun, Claudia Cummins and Dave Baillie et al. for their assistance, and Alan Coulson et al. for help with mapping clones. Supported by a Research Reorientation Associateship (NSERC Canada) to SS, and an NSERC Operating grant to BMH.
-
[
International C. elegans Meeting,
2001]
In vertebrate cells the K-Cl cotransporter has the capacity to reduce cellular concentration of K and Cl. This plays several documented roles: regulation of cytoplasmic chloride concentration, reduction of cell size in development, defense against cell swelling under hypoosmotic and isoosmotic conditions. The last, which includes the defense against swelling during moderate warming due to hyperactivity of the Na-K pump, has received the least attention but may be the most physiologically relevant. The gene sequences for two mammalian isoforms of this cotransporter were identified in 1996 and at that time two similar sequences were recognized in the C. elegans database (KO2A2.3, "KO2", on chromosome 2, and R13A1.2, "R13", on chromosome 4). Both C. elegans isoforms are expressed,as demonstrated by their presence in EST's and cDNA libraries. K02 has been shown to have K-Cl cotransport activity when transfected into mammalian cells. We have carried out dsRNAi blockages against both of these genes, singly and in combination, and found that about a third of the injected parents produced progeny that exhibit reduced survival at elevated temperature (33degC and 37degC) and increased sensitivity (seen as immobilization) to isoosmotic challenge induced by exposure to 100 or 200 mM ammonium chloride in M9 medium. A deletion mutant of K02 obtained from Gary Moulder at U. Oklahoma has not shown a phenotype by these tests and is now being used as a target for RNAi knockout of the R13 gene. GFP's expressed from upstream sequences of the two genes indicate that both isoforms are consistently expressed in pharyngeal muscle. The extent to which either is expressed elsewhere is still being determined. These studies may lead to establishing the role of K-Cl cotransporters in the whole organism and lay the foundation for a genetic analysis of their control
-
[
Worm Breeder's Gazette,
1987]
The reason worms don't fall off an upside-down plate is surface tension. The force pressing the worm to the agar surface is roughly the surface tension of water times the perimeter of a sagittal section: 10+E5 -10+E6 times the weight of the worm for a young L1. Because of this, I thought worms might be able to survive high-speed centrifugation. Since Gary Ruvkun bet me ten dollars they wouldn't, I did the experiment. The worms were very young L1s prepared by hatching eggs in the absence of food. I first spun some at 100,000G (32,500 rpm in an 80Ti rotor). The spin was as short as I could make it: 5 min acceleration, a few seconds at speed, and 5 min deceleration. The worms were unaffected. About half of them thrashed in liquid, all looked normal in Nomarski, and all grew up to healthy, active adults and produced large broods in the normal time. Next I spun some more worms at 460,000G (70,000 rpm, as fast as our centrifuge would go). These worms were all clearly abnormal in Nomarski. There were usually empty spaces between the buccal cavity and the head cuticle, and the tail often had similar holes. In some the viscera pulled away from the cuticle in other places, too. Many individual cells were killed. Most of these worms arrested as L1s, though some continued to move. But some of them grew up to become healthy adults and produce progeny. Spinning at high speed stresses worms in three ways: there are forces flattening them against the bottom of the tube (like the surface tension forces on a plate), a very high static pressure that will affect chemical reactions (e.g. dissolution of gases, as in the bends), and forces causing components of the worm to sediment with respect to each other. Judging from the Nomarski result, it might be the last of these that bothers the worms most, though they are very resistant to all three.