Kreutzer MA et al. (1994) Worm Breeder's Gazette "Expression Studies of the Putative Caenorhabditis elegans cyclin A and B Genes"

Richards JP, Bennett KL, Kreutzer MA

[Worm Breeder's Gazette, 1994]

Expression studies of the putative Caenorhabditis elegans cyclin A and B genes Monique A. Kreutzer, James P. Richards and Karen L. Bennett, University of Missouri-Columbia, Columbia, Missouri 65212. We have cloned putative A, B1 and B2 cyclins from C. elegans. Cyclin B1 and B2 are single copy genes while cyclin A belongs to a multigene family. Cyclin A and B2 each correspond to single transcripts while cyclin B1 recognizes three transcripts. The C. elegans temperature sensitive germline defective mutants fem-1, fem-3 gf) and glp4 were used to analyze cyclin germline expression. When hybridizing C. elegans cyclin A or B2 to the mutant RNAs that produce no oocytes, the cyclin A and B2 messages are significantly reduced. When normal numbers of oocytes are produced, the cyclin A and B2 messages are at wild type levels. Therefore, the cyclin A and B2 transcripts appear to be primarily maternal with some somatic component. Consistent with a somatic component, cyclin A and B2 cDNAs cross-hybridize to Ascans intestinal poly A+ RNA. Hybridization of the cyclin B1 cDNA to these germline defective RNAs suggest that the cyclin B1 transcripts are differentially expressed in the germline. The two larger cyclin B1 transcripts appear to be primarily maternal and the smallest transcript appears to be sperm- specific. The observation that two polyadenylation signals are present in the cyclin B1 3'UTR, as well as results using cyclin B1 probes from specific regions of the B1 3'UTR, suggests that two of the three cyclin B1 transcripts are produced by polyadenylation a~ different sites. There is precedence in mice and flies for producing tissue-specific transcripts by different use of the cyclin B 3'UTRs. It has been shown in mouse that cyclin B1 hybridizes to four differentially expressed transcripts, one of which is testis- specific. Two of these mouse transcripts differ in the choice of polyadenylation sites and therefore in the lengths of their 3'UTRs (Chapman and Wolgemuth (1992) MoL Repro.Dev. 33, 259-269). In Drosophila, the cyclin B cDNA detects two female-specific transcripts that are made by splicing of a region of the cyclin B 3'UTR (Dalby and Glover (1992) Development 115, 989-997). In summary, our initial findings show that the C. elegans cyclin As and Bs are highly expressed in the germline and also suggest that the three cyclin B1 transcripts are differentially expressed in oocytes and sperm.

Danielle Thierry-Mieg et al. (2003) Worm Breeder's Gazette "www.wormgenes.org , a new gene centered database"

Vahan Simonyan, Michel Potdevin, Mark Sienkiewicz, Yuji KOHARA, Jean Thierry-Mieg, Danielle Thierry-Mieg, Tadasu SHIN-I

[Worm Breeder's Gazette, 2003]

Wormgenes is a new resource for C.elegans offering a detailed summary about each gene and a powerful query system.

Yook K et al. (1995) Worm Breeder's Gazette "A Screen For Lethal Synaptic Function Mutations."

Yook K, Jorgensen EM

[Worm Breeder's Gazette, 1995]

A screen for lethal synaptic function mutations Karen Yook and Erik Jorgensen Universitv of Utah, Dept. of Biology. UT 84112 Our goal is to identify proteins that function at all synapses. These include proteins that are involved in svnthesis, packaging, transport and the release of neurotransmitters into the synaptic cleft, as well as proteins involved in the recycling of synaptic vesicles. Initially we screened for mutants that behaved as if they were defective for both GABA and acetylcholine function: 11 genes were defined by this approach. One drawback with such a screen is that it will fail to identify lethal mutations. We have developed a screen which can potentially identify lethal mutations that interfere with neurotransmission. Aldicarb is a pesticide that inhibits the breakdown of acetylcholine in the synaptic cleft. The resulting accumulation of acetylcholine in wild-type worms results in paralysis. whereas worms lacking or having reduced acetylcholine transmission are resistant to the drug and continue to move. It was noted that many of the genes uncovered in the behavioral screen conferred resistance to aldicarb as double heterozygotes with n2813. an allele of unc-13 (for example unc-13/+: snt-l/+). Apparently neurosecretion is very sensitive to the dosage of proteins required for this process. This sensitivity allowed us to screen for haploinsufficient mutations in a background sensitized by the unc-13 mutation. We mutagenized wild-type males and crossed them to a strain containing unc-13 (n2813) (see Figure below). In the Fl we picked aldicarb-resistant individuals. Lethal mutations in genes that are essential to synaptic function are maintained as aldicarb resistant heterozygotes. We are interested in two classes of mutations: The first class are lethal mutations, identified as strains that are resistant to aldicarb when unc-13 is heterozygous, and also throw lethal progeny. The second class are homozygous viable aldicarb-resistant mutations that can be separated from the unc-13 mutation. Of 2336 genomes scored. we uncovered 62 possible lethal mutations and 61 homozygous viable aldicarb-resistant mutations. In the homozygous viable class we found both uncoordinated and wild-type mutants. All of these strains have satisfied preliminary criteria for synaptic function mutants and we are confident that our screen will be successfill in uncovering genes of interest. One mutation pulled out of the modified screen has been shown by noncomplementation to be an allele of unc-26, mutations of which are known to be resistant to Aldicarb (J. Rand, personal comm.).

Edgar RS (1975) Worm Breeder's Gazette "the shipping and handling of nematodes."

Edgar RS

[Worm Breeder's Gazette, 1975]

Many of us have totally abandoned wooden sticks for transferring worms. Instead a short (1 - 1 1/2') piece of platinum wire (32 gauge) is sealed in a pasteur pipette holder. Platinum is soft and a sharp hook or spatular end can be shaped with a razor blade and forceps. Various 'tools' of varied shape can be manufactured. Such implements are readily sterilized in a micro-burner. I have had some success shipping worms in letters. Three or four small pieces of filter paper are placed on a piece of aluminum foil. The filter papers are saturated with a buffer washate from a starved plate. The foil is folded several times to create a seal. The recipient places the filter papers on a plate or washes them in buffer.

Gruidl ME et al. (1995) Worm Breeder's Gazette "The Adult Sterile Mutant, let-545, Is Likely To Be a Mutation in glh-1, a Putative P-Granule Component."

Gruidl ME, McKay S, Bennett KL, Rose AM

[Worm Breeder's Gazette, 1995]

The adult sterile mutant, let-545, is likely to be a mutation in glh-1, a putative P-granule component.

Lewis JA (1988) Worm Breeder's Gazette "referencing, crossreferencing, and creating reference lists with Microsoft Word."

Lewis JA

[Worm Breeder's Gazette, 1988]

I've written a little primer on how to use Word (version 3.0 or higher) to automatically generate references (reference number or author-year citation) and cross-references (to page number) within a document using a combination of Word's merge function and table of contents generation facility. Also included in the article are directions for using reference information stored in a database to automatically generate a reference list according to a given style using Word's merge function. The utility of all this is likely to be superseded by future versions of Word and commercially available add- on programs. However, if you are interested, drop me a line and I'll send you a copy of my article.

Way JC et al. (1986) Worm Breeder's Gazette "a simple method for decontaminating worm stocks."

Way JC, Chalfie M

[Worm Breeder's Gazette, 1986]

In the past, we have used the standard method for decontaminating stocks, which involves washing worms off a plate, spinning them in a centrifuge, etc The following procedure is much quicker and requires fewer worms. Also, it can be used to directly decontaminate the progeny of a mating and thus accelerate strain constructions. Using a platinum wire with a glob of bacteria on the bottom, pick up several gravid adults and/or eggs from the contaminated plate and transfer them to a seeded plate. Then put a drop of the usual sodium hypochlorite solution (2-4% NaOCl, .4M NaOH) onto the eggs. About half of the eggs will hatch. Occasionally, a very tough bacterial contaminant will survive to form a few colonies, so it is wise to transfer the survivors on the following day.

Weaver DC et al. (1995) Worm Breeder's Gazette "A Nob Allele at the unc-62 Locus."

Edgar LG, Wood WB, Weaver DC

[Worm Breeder's Gazette, 1995]

A Nob Allele at the unc-62 Locus

Driscoll MA et al. (1992) Worm Breeder's Gazette "A Hot-Spot for Mutations in a Cys-rich Domain of mec-4 Includes a Site Which,When Mutated, Can Suppress a Dominant Death-Inducing Mutation in Cis"

Driscoll MA, Hong K

[Worm Breeder's Gazette, 1992]

A Hot-spot for Mutations in a Cys-rich Domain of mec-4 Includes a Site Which, When Mutated, Can Suppregg a Dominant Death inducing Mutation in Cis Kyonsoo Hong and Monica Driscoll. Department of Molecular Biology and Biochemistry, Rutgers University, Center for Advanced Biotechnology and Medicine, 679 Hoes Lane, Piscataway, N.J. 08855 (908) 463-5193

Hecht RM et al. (1980) Worm Breeder's Gazette "ontogeny of poly(A)+ RNA in C elegans."

Gossett LA, Hecht RM, Jeffery WR

[Worm Breeder's Gazette, 1980]

We have examined the titer of poly(A) RNA in squashes prepared from oocytes and variously staged embryos of the nematode Caenorhabditis istol (N2) by In situ hybridization with a [3H]-poly(U) probe. Control experiments demonstrated that this probe interacts specifically with poly(A) sequences and is capable of detecting poly(A) RNA. Using this method and saturating concentrations of the probe, it was shown that isolated oocytes and embryos up to the 125-cell state exhibited substantial levels of poly( A) RNA in their cytoplasms but almost no activity within their nuclei. Nuclear poly(A) RNA first appeared at the 90-cell stage and afterwards increased at a linear rate through the 550-cell stage. The titer of total embryonic poly(A) RNA also increased at a linear rate up to larval hatching. The results suggest that the major transcriptional effort for poly(A) RNA begins at the 90 to 125-cell stage of C. elegans embryogenesis and that the poly(A) RNA present exclusively in the cytoplasm prior to this time is of maternal origin.