Bertrandy S et al. (1998) Worm Breeder's Gazette "Looking in C.elegans for the function of SMN, the Spinal Muscular Atrophy disease gene."

Thierry-Mieg D, Lefebvre S, Munnich A, Bertrandy S, Powell-Coffman JA

[Worm Breeder's Gazette, 1998]

We are grateful to the genome project for sequencing cosmid C41G7 (chromosome I, +3.7 cM), which contains an ortholog of the SMN (Survival of Motor Neuron) gene. Mutations of the telomeric copy of the duplicate SMN gene leads in human to the second most frequent fatal autosomal recessive disease of childhood, spinal muscular atrophy or SMA (Lefebvre et al., Cell 80, 155-165, 1995). The illness is characterized by degeneration of motoneurons of the spinal cord. A tight correlation between clinical severity and SMN protein level was demonstrated in SMA patients and a strong protein expression was observed in the spinal motor neurons from control individuals (Lefebvre et al., Nature Genetics 16, 265-269, 1997). The function of the SMN gene product is unknown. The SMN protein appears to form a complex with spliceosomal snRNP proteins (Liu et al., Cell 90, 1013-1021, 1997) and to participate to spliceosomal snRNP biogenesis (Fischer et al., Cell 90, 1023-1029, 1997). We wish to characterize the nematode gene, its functions and regulations, in the hope that this knowledge may help device therapeutic approaches for the 1/6000 unfortunate homozygotes. No cDNA clone was available when we started the project a year ago. We isolated, cloned and sequenced the full length cDNA. It measures 0.7 kb, does not appear to be transpliced and exactly matches the genefinder prediction of the coding region (4 exons, 197 aa). We then generated transgenic strains expressing a translational fusion of GFP to the SMN genomic sequence, including the SMN promotor (0.53 kb of upstream sequence, touching the nearest upstream gene, which encodes a ncd-like kinesin). The SMN gene is fused to GFP-NLS (vector pPD95.70) 8 aa before the stop codon. Out of 44 Rol, only 2 transmitted. These transgenics fluoresce at very low levels in most tissues, with two dozen neurons (not motoneurons) a bit brighter than the other cells. In the course of these experiments, we observed that injection of the GFP fusion construct and the pRF4 Roller plasmid in the ratio of 50: 150 leads, 12 to 16 hours after the injection, to irreversible body muscle paralysis of the injected worms. The injectees look healthy, eat and lay between 60 and 140 eggs per hermaphrodite but move only the head. The eggs laid will either die as embryos (85%), or hatch as very short Lls, with a big head and an often knobbed tail (15%). We are intrigued by the paralysis which directly affects the injected animals and wonder if other cases of such "diffusible" effects were ever observed ? RNA interference experiments were also conducted. The injectees generated a few small round eggs, some dead embryos and a proportion of larvae that died as L2 or L3 and were either Unc (not paralysed, but uncoordinated like the acetylcholine receptor mutants) and/or strongly dumpy (like dpy-17). But the most striking failure was in the differentiation of the germ line. Defects ranged from empty gonads (no or few germ line cells, but normal somatic gonad), to Glp-l like (only a few sperms), to small non growing ovocytes (vitellogenin accumulates in the intestine, a few tiny round eggs may be laid, none survives). A large number of animals also had misguided gonad(s), like Unc-5. Based on these phenotypic informations, we are selecting new alleles in standard mutagenesis screens, using mnDfl11 as uncovering deficiency and transgenics as wild type duplication. We are also looking at previously identified mutants in the area. In parallel, we started biochemical experiments. Rabbit antisera were raised against three peptides, and we are currently using them in immunoblots, and will try them soon in immunofluorescence.

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.

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.

Stein JA et al. (1984) Worm Breeder's Gazette "a computer program for organizing a clone collection."

Emmons SW, Stein JA

[Worm Breeder's Gazette, 1984]

We have written a computer program for storage and retrieval of information about the recombinant DNA collection held by a laboratory. For each clone, information such as clone name, cloning vector, cloner, date cloned, notebook page, storage location, etc., is entered. The cloned insert is identified by a serially-assigned fragment number and by a restriction map. Wherever a given fragment occurs in the collection it is identified by its fragment number, and all restriction mapping data for that fragment is held together in a single storage location and kept updated. Data regarding interrelationships among fragments in the collection, such as which are subclones of others, are also held. The stored information can be searched and listed in a variety of ways. The program is modular in design and can be readily modified and expanded. It is written in Basic for a Dec RT-11 operating system ( PDP-11 computer or equivalent) and should be easily adaptable to other systems. We encourage other laboratories to use it. The program anticipates multiple laboratory use by prefixing each fragment number with a laboratory number. By this means each cloned segment of the genome is given a unique name. Eventually there will undoubtedly be a need for a centralized listing of information about cloned fragments of the C. elegans genome. We suggest that a centralized listing could be a subset of the information stored in laboratory based listings such as this one. For a free interchange of information among laboratories and between laboratories and a centralized data base to be possible, a uniform system of nomenclature and computerized format will have to be adopted. We have written this program in order to gain some experience with the use of a laboratory based data bank that hopefully will be helpful in designing a widely used system. Please contact us if you are interested in using this program. We would be glad to help you adapt it to your computer system.