De Stasio E et al. (1994) Worm Breeder's Gazette "Mutagenesis of C. elegans Using N-ethyl-N-nitrosourea."

De Stasio E, Stanislaus D, Lephoto C

[Worm Breeder's Gazette, 1994]

Mutagenesis of C. elegans using N-ethyl-N-nitrosourea Elizabeth De Stasio, Dinesh Stanislaus and Catherine Lephoto. Department of Biology, Lawrence University, Appleton, Wl 54911

Creg Darby (2002) Worm Breeder's Gazette "Plague? Bah! Mutants resistant to the bubonic plague biofilm"

Creg Darby

[Worm Breeder's Gazette, 2002]

Bubonic plague is spread by the bites of fleas infected with Yersinia pestis bacteria. When worms are exposed to Y. pestis, sticky biofilms appear on their heads. The biofilms cover mouths and impair feeding, which in the case of larvae delays or arrests growth. Bacterial genes required for flea infection are also required to form worm biofilms, suggesting that the plague-flea interaction can be modeled with C. elegans (1).

Albertson DG et al. (1986) Worm Breeder's Gazette "progress in mapping genes."

Sulston JE, Fishpool RM, Albertson DG, Coulson AR

[Worm Breeder's Gazette, 1986]

Cloned genes are readily and routinely mapped to linkage groups of C. tu hybridization using the corresponding cosmids as probe DNAs. Our progress is summarized below. A few genetic markers (lower case letters above the line) have been included for reference. Uppercase letters above the line represent cloned DNAs corresponding to genes that have been sent to Cambridge. Genes designated by uppercase letters within error bars below the line are those mapped by in situ hybridization.

Papp A (1992) Worm Breeder's Gazette "Results of Worm Plate Survey"

Papp A

[Worm Breeder's Gazette, 1992]

After tabulating the results of the Worm Plate Survey. we have come up with some interesting results. Most notably. the high variability in prices that labs are paying for their plates, even for the exact same plates from the same supplier, and the fact that most plates are marked up considerably over the actual cost. The replies can be separated into 4 categories: Labs that get plates from Fisher ($29-$58). but wish they had non-vented plates Labs that get non-vented plates via Applied Scientific (~$38) Labs that get plates from Falcon (vented) or Nunc (non-vented) and pay much more Most labs' plates were "slipable" or "semi-stackable", but all labs wanted plates that stack well for easy manual pouring, seeding, carrying, and using. Everyone wanted plates with shallow lids such that the bottoms can be lifted out of the tops for inverted use. Some labs expressed an interest in plates slightly smaller than "60 mm". That number is in quotes because all of the companies' plates have bottoms smaller than 60 mm (e.g. Fisher -54 x 14 mm). We have negotiated with the plastic companies that really make the plates for Fisher, Applied Scientific, etc. (that actually just resell them to you). I have come to the conclusion that we can provide you with better worm plates, the same worm plates cheaper, or in most cases better worm plates cheaper. This is true for every lab. The bottom line is that we can get you top quality non-vented "60 mm" plates (like Applied Scientific's, except fully stackable) for about $29 per 500 case INCLUDING shipping depending on your usage and how many cases you can receive at one time. Several labs have found the non-vented plates last longer without drying out or getting contaminated, compared with normal vented plates, so you should save that way, too. We offer full service shipping (e.g. standing orders and same-day telephone orders, free. Similarly low prices are available on 100 mm and 150 mm plates that exceed industry standards for flatness (reducing media usage) and clarity. The 100 mm are about $27 per 500 case plus shipping; The 150 mm dishes (good for DNA & RNA preps and library platings, with more than 2.25x the surface area of 100 mm dishes) are made thicker and deeper than industry standards and are about $21.50 per 100 case plus shipping. The shipping charge is very low for labs, or groups of labs in one city, that can take delivery of many cases in a single shipment. You can even suggest that your stockroom order plates from us. Call us for an exact price quote depending on your usage and how many cases you can receive at one time. In any case, we'll work things out to save you money. In the future, we can offer inexpensive 35 mm dishes if the community at large can order about 2000 cases per year, so let me know about your needs for other sizes. The response was very mixed about pre-poured plates. We may set that up later, but for now we can help the most by saving you lots on empty petri dishes (and later, maybe media .supplies). We are happy to send out free samples so you can examine the dishes. If we haven't contacted you yet, just give us a call. Respondents: 38 (including 5 anonymous) "Winners": Horvitz = 550, Meyer = 400, Thomas = 400, Greenwald = 300 200-299 cases 8 labs 100-199 cases 7 labs 4-99 cases 19 labs Highest price per case: US = 118.75, Canada = $117 (non-vented) Lowest price per case: US = $29, Canada = $25 (vented) Farthest away response: Malta! No responses from MRC or anyone else in Europe or Asia. It is possible that we can save money and/or provide better plates for these labs, including, shipping, too. Let us know.

Ouazana R (1979) Worm Breeder's Gazette "some enzymatic comparisons between Bergerac and Bristol strains of C elegans."

Ouazana R

[Worm Breeder's Gazette, 1979]

To compare enzymatic activities of Bergerac and Bristol strains of C. elegans, we used the microsystem ' Api Zym ' focused by D. Monget ( 1978) and produced by the french firm API SYSTEM. Some of these tests, not yet commercialized, were a gift of API SYSTEM. The technique consists in microdishes which are impregnated with substrate and buffer ( PH is given in the table ). We introduced 0.1 ml of biological suspension ( a known number of adult worms sonicated in buffer and resuspended in distilled water after homogenization ) in the microdishes which were incubated at 37 C for 4 hours. Bacteria were grossly eliminated by washing; Afterwards their concentration was too low to give response with Api Zym. The reactions were revealed colorimetrically by 4 /oo ' Fast blue BB '. The intensity of the reaction is proportional to enzymatic activity and is noted from - to +++++ . A reaction noted +/- refers to a low but detectable response. To give reliable results, all the tests were read by the same experimenter and enzymatic activities in Bergerac and Bristol strains were performed at the same time. To date, the tests were carried out on 64 enzymatic activities. Most of the enzymes tested were hydrolases ( phosphatases,esterases, aminopeptidases, glycosidases, arylsulfatases ) and some were dehydrogenases. Some of the enzymatic activities tested seem to take a prominent part in postembryonic development. It's the case of Leucine arylamidase, which plays an important part in molting ( Rogers 1965, 1977 ), and could be also the case of glycyl-L-proline arylamidase which is implicated in the metabolism of collagen. The results are summarized in the following table. The two strains show similar enzymatic spectra, but differences can be noted, which are localized essentially on : - Trypsin ( n 8 ) - gamma L-glutamate transpeptidase ( n 38 ) - Glycyl-glycine arylamidase ( n 42 ) - L-threonine arylamidase ( n 54 ) - N-CBZ-glycyl-glycyl-L-arginine arylamidase ( n 56 ) Other differences, in a less degree, but yet detectable, are observed on: - Alkaline Phosphatase ( n 1 ) - L-histidine arylamidase ( n 33 ) - L-leucyl-glycine arylamidase ( n 45 ) - L-ornithine arylamidase ( n 51 ) - L-serine arylamidase ( n 53 ) All the differences observed in this table have been verified in two repetitions, which have similar results. But it could be argued that these differences could be due to nutritional adaptation, because these two strains, xenically cultured on the A1 medium ( for thirty years in case of Bergerac, and one year in the case of Bristol ) are associated with a different bacterial complex. [See Figure 1]

Plasterk RHA (1988) Worm Breeder's Gazette "A Tc1 homologue in Drosophila."

Plasterk RHA

[Worm Breeder's Gazette, 1988]

Recently a homology was found between the TcA gene in Tc1 and an ORF in the Drosophila low repetitive sequence HB (Dr. S. Henikoff pers. comm. and manuscript in prep.). If one assumes that the Tc1 TcA gene encodes the transposase of the element, then it would follow that the target sequence of this enzyme (the ends of Tc1) could be homologous to those of the HB sequences as well. Therefore I compared the ends of the two elements. Since HB has not been shown to jump, the exact ends of the element are not defined. I looked at 8 ends of 4 elements (1), and took the point up to where they are identical or 7 out of 8 identical to be the assumed end of HB. In the figure the assumed end of HB and the end of Tc I (2) are compared. [See Figure 1] As can be seen they have 16 out of 24 basepairs in common. This is sufficient to conclude that Tc1 and HB elements are related transposable elements (or in the case of HB perhaps relics of transposable elements). To my knowledge this is the first case of similarity between DNA transposons of not directly related species. I'd like to thank Dr. S. Henikoff for communicating his observation prior to publication.

White JG et al. (1976) Worm Breeder's Gazette "studies on the structures of the ventral cords of developmental mutants."

Southgate E, Anness M, Sulston JE, White JG

[Worm Breeder's Gazette, 1976]

We have been studying the ventral cords of two mutants which fail at different stages in the period of post-embryonic development which occurs just prior to the L1 moult. In one mutant, E1348 isolated by J. E. Sulston, the precursors, which would normally give rise to 5 neurones and one hypodermal cell, do not divide while in the second, E1414, isolated by H. R. Horvitz, only the last rounds of division are variably blocked. In both these mutants the structure and organization of the juvenile set of motor neurones is the same as that in the wild-type adults. The 'precursor' cells in H86 generally turn out to be motor neurones, although one case has been seen where one had become a hypodermal cell which had fused with the hypodermal syncytium. The neurone-like cells have many more branches than normal motor neurones and show some of the characteristics of the individual neurones that would normally be produced from these cells, such as the characteristic dendritic spines of the class D motor neurones. In the case of E1414, some of the neuroblasts that would normally give rise to a type A and a B or an AS and a VD motor neurone fail in this terminal division. These cells take on the characteristics of their posterior daughters i.e. a class B and a class VD, both in terms of their morphology and synaptic input. Those neuroblasts that divide normally produce normal looking progeny cells.

Lee JH et al. (1992) Worm Breeder's Gazette "unc-101, a Negative Regulator of the Vulval Induction Pathway, Encodes a Clathrin Adaptor Protein"

Jongeward GD, Lee JH, Sternberg PW

[Worm Breeder's Gazette, 1992]

unc-101 is a gene required in many different tissues for survival, defecation, development of some neurons, and normal induction of the vulva. While unc-101 mutants do not have any vulval defect, they have an ability to suppress the vulvaless phenotype of some weak alleles of let-23 .They also suppress strong alleles of lin-2 ,lin-7 ,and lin-10 . We cloned the unc-101 gene, and determined the genomic structure and the cDNA sequence. The gene has 7 exons and 6 introns. The cDNA sequence predicts a protein of 422 amino acids. A database search identified a strong homology to the rat AP47 ,the medium chain of Golgi-associated clathrin adaptor protein complex AP- 1. The amino acid sequence identity between unc-101 and AP47 is 79%. Intensive biochemical studies have been done on the mammalian adaptor protein complexes. The coated pits of the plasma membrane and Golgi consist of clathrin and adaptors interacting with receptors on the membrane. There are two different adaptor protein complexes, AP-1 and AP-2. AP-1 is localized in trans- Golgi network and is involved in protein sorting and regulated exocytosis. AP-2 is localized on the plasma membrane and is involved in endocytosis of receptors. The medium chains of these adaptor complexes, called AP47 and AP50 respectively, show limited homology to each other (40% identity). A C. elegans homolog of the medium chain of AP-2 was identified by the sequencing consortium (WBG vol.12, No.2, p26 ). Given very high sequence homology to each other, the mammalian and C. elegans homologs may well have a conserved function. There are many possible roles for unc-101 at the subcellular level including: 1) unc-101 might be involved in the retention of let-23 molecules after their biosynthesis. Then unc-101 mutant cells could have more receptors on their surface. 2) unc-101 might be involved in sorting of lysosomal enzymes required for degradation of signal transduction machinery. Then the unc-101 mutant cells could have more signal transduction. In either case, unc-101 should be expressed in the vulval precursor cells. 3) unc-101 might be responsible for the regulated secretion of the signal molecules. Then the unc-101 mutants could have more signal for vulval induction. In this case, unc-101 should be expressed in the anchor cell. 4) unc-101 might be involved in the regulated secretion of negative regulators of vulval differentiation (e.g. proteases required for degredation of extracellular signal). In this case, unc-101 could be expressed in the hyp7 cells.

Hall DH et al. (1975) Worm Breeder's Gazette "Synaptic Anatomy of the C. elegans"

Hall DH, Russell RL

[Worm Breeder's Gazette, 1975]

Two young adult animals have been serially sectioned from the tip of the tail forward through the anterior end of the preanal ganglion. The 9 cells of each lumbar ganglion and the 12 cells of the preanal ganglion can be reproducibly identified on the basis of a set of morphological features, including cell body position, direction and extent of fiber projection, fiber size, and cytoplasmic appearance. A total of at least 6 bilaterally homologous pairs can be identified among the lumbar cells. Virtually all the synaptic contacts occur in the preanal ganglion; very few are found in the lumbar ganglia. However, the most prominent participants in these synapses are lumbar cells and a few cells whose bodies lie anterior to the sectioned region; the preanal ganglion cells themselves are relatively minor participants. A complete connectivity matrix has been constructed for both animals, involving about 90 synapses in each case. Certain cells make repeated contacts with one another (up to 13 contacts) in both animals. Other instances of non-reproducible synapses are found, usually involving one contact in one animal and none in the other. Homologously paired cells make similar sets of synaptic contacts. Most (~85/90) of the contacts are diadic, with one presynaptic cell and two postsynaptic ones. Several instances of 'multiple routes of information flow' are found, in which cell A sends to cell C in two ways, both by direct synaptic contact and through an intervening cell, B. No self-synapses are observed, but sensory cells frequently synapse onto their bilateral homologs. One case of reciprocal synapse formation is found. Most of the contacts can be included in a simple wiring diagram by which information flows from sensory cells through multiple routes to converge on a pair of interneurons that apparently constitute one of the major outputs of the system.

Sattelle DB et al. (1996) Worm Breeder's Gazette "acr-2, acr-3: a pair of linked genes on chromosome X encoding non-alpha nicotinic acetylcholine receptor subunits (ACR-2, ACR-3) capable of functional co-expression in Xenopus oocytes with UNC-38 to yield small amplitude cationic currents"

Sattelle DB, Fleming JT, Barnard EA, Squire MD, Baylis HA, Torne C, Matsuda K

[Worm Breeder's Gazette, 1996]

Several putative nicotinic acetylcholine receptor (nAChR) subunit clones were isolated by screening a lambda library of Caenorhabditis elegans genomic DNA with a probe derived from the Drosophila melanogaster ard gene (a non-alpha nicotinic acetylcholine receptor subunit clone). Studies on two of these loci, acr-2, acr-3 and their gene products ACR-2 and ACR-3 are described. acr-2 is located between sup-7 and unc-6 on the X chromosome. The cDNA encoding ACR-2 was sequenced and shown to possess many of the conserved features of vertebrate and invertebrate non-alpha nicotinic acetylcholine receptor subunits. To investigate the functional expression of this subunit, the corresponding cRNA was produced and micro-injected into the cytoplasm of Xenopus oocytes. When expressed alone, ACR-2 was not able to generate levamisole-gated channels. When co-expressed with a C. elegans a subunit (UNC-38), which is itself unable to form functional homo-oligomers, ACR-2 contributed to the formation of a functional channel that was blocked by mecamylamine1. Recently, similar findings have been obtained for the product of acr-3, another non-alpha nAChR subunit encoding gene located 281bp downstream from acr-2 on the X chromosome and in the same orientation. In this case, ACR-3 encoding DNA was injected into Xenopus oocyte nuclei. As in the case of ACR-2, small cationic currents were detected in the presence of 100mM levamisole and these were blocked by mecamylamine (1mM) and d-tubocurarine (10mM). Studies on the regulation of expression of these linked non-alpha subunits are underway. The functional expression studies of nematode nicotinic acetylcholine receptors reported here support the interpretation that the differentiation between alpha and non-alpha subunits dates back to the earliest stages of metazoan evolution.