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[
Nature,
1976]
The study of mechanisms affecting the rate of ageing can be facilitated by naturally occurring phenomena, innate to some organisms, that enable the species to retard its ageing rate and extend its life span. Such a phenomenon exists in certain species of nematode. Larval forms of the free-living soil nematode, Caenorhabditis elegans, possess the ability to enter a semi-dormant, quiescent state referred to as the dauer larval stage (German, "enduring" larva). Newly hatched larvae of C. elegans undergo four larval stages (L1-L4) punctuated by moulting. If larvae are starved, they will enter the dauer state during the second larval moult. At this time, the old cuticle is replaced by a special, relatively impermeable cuticle unique to dauer larvae.
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[
Dev Biol,
1979]
The temperature-sensitive sex transformer
tra-2(
b202)II of the nematode Caenorhabditis elegans causes the tranformation of genotypically hermaphrodite worms into phenotypic males and sterile intersexes at restrictive temperature. In this note, we show that the entire gonad structure is transformed and that oocyte development is autonomous of the form of the gonad and of the presence of a cellular sheath. Four oocyte-specific proteins are present in male intersexes that produce oocytes but are lacking in transformed males and hermaphrodite intersexes
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[
Mech Ageing Dev,
1977]
The free-living nematode Caenorhabditis elegans is an excellent experimental system for the study of aging. The present study identifies some of the major biological and environmental factors influencing life span as a prelude to more detailed genetic and biochemical analyses. Life span can be altered during any part of the life cycle by a change in either temperature or food concentration. Parental age and parental life span both have relatively small effects on progeny life span. The nematode accumulates fluorescent pigment resembling lipofuscin, and becomes less sensitive to ultra-violet radiation as it
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[
Mech Ageing Dev,
1983]
The free-living nematode Caenorhabditis elegans is used as a genetically manipulable experimental system for the study of aging. Utilizing a temperature-sensitive sterile strain with a normal life span, a method is described for the isolation of mutant strains with significantly increased life spans. Eight mutant strains were isolated each having increased life spans. Two mutant strains were spontaneous dauer formers, accounting for their increased longevity. Another was chemotaxis-defective, causing reduced food intake which could account for its increased life span. Five mutants suffered from varying degrees of paralysis affecting their rate of pharyngeal pumping and food ingestion. The high correlation of the decreased rate of food ingestion of these mutants with their increased longevity is interpreted as indicating that the increased longevity is most likely due to reduced caloric intake. These results appear to indicate that specific life span genes are extremely rare or, alternatively, life span is controlled in a polygenic fashion.
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[
Mech Ageing Dev,
1983]
Analysis of DNA from the nematode Caenorhabditis elegans demonstrated a number of significant age-correlated changes. The number of single- strand breaks as assayed by an in vitro assay procedure using Escherichia coli DNA polymerase I increased significantly with age. There was also an exponential increase in the amount of 5- methylcytosine in C. elegans DNA as the worm matured and aged. Furthermore, DNA isolated from older worms exhibited reduced transcriptional capacity when assayed in a HeLa cell in vitro transcription system. Finally, a biological assay to determine age- correlated changes in the DNA of aging sperm demonstrated a significant reduction in the capacity of the sperm to support zygotic development as the age of the male increased. These findings demonstrated significant age-correlated alterations and modifications occurring in the DNA template of the nematode, and their implications to the aging process are discussed.
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[
Dev Biol,
1976]
The small free-living nematode Caenorhabditis elegans is usually found as a hermaphrodite, but occasionally true males appear in the population. This study provides an account of gonadogenesis in the normal male and in a mutant that is a temperature-sensitive sex transformer. Male and hermaphrodite gonads develop from morphologically identical primordia. The small primordial gonad lies on the ventral side of the worm in the coelomic cavity. The gonadial primordium contains four nuclei at parturition. As this primordium develops in a hermaphrodite, it produces a double-armed, mirror symmetrical gonad that produces first sperm and then eggs. In the male, however, this primordium develops into an asymmetrical structure composed of a ventrally located testis, a loop region, a seminal vesicle, and a vas deferens. The male gonad presents a linear sequence of nuclei in successive stages of spermatogenesis with a mitotic region in the testis, followed by clearly distinguishable stages of meiosis throughout the loop region to the seminal vesicle. A temperature-sensitive sex transformer mutant, tsB202, has been isolated. tsB202 carries an autosomal recessive mutation in linkage group II that at restrictive temperature transforms an XX hermaphrodite into a phenotypic male, complete with a normal male gonad and vestigial external genitalia. These transformed males are classified as pseudomales because they do not exhibit mating behavior. Temperature shift experiments have determined the specific temporal sequences of gonadogenesis, oogenesis, and spermatogenesis. Proper manipulation of the temperature regimen causes the production of intersexes. In one intersex, a male gonad complete with sperm, seminal vesical, and vas deferens also contains oocytes. In another intersex produced by the complementary temperature shift, a hermaphrodite-shaped gonad develops that produces only sperm and no oocytes.
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[
Dev Biol,
1982]
The major protein found in nematode sperm exhibits a distinct pattern of developmental regulation. In the nematode Caenorhabditis elegans, the synthesis of the major sperm protein (15K) begins with the onset of spermatogenesis in both the male and hermaphrodite. Both spermatogenesis and 15K synthesis continue for the life of the male while in the protandrous hermaphrodite the major sperm protein is synthesized only during the fourth larval stage. Inhibitor studies using actinomycin D and a-actinin as well as Northern blot analysis have shown that the primary regulatory mechanism of this gene is at the transcriptional level. Recombinant molecules have been selected bearing the 15K genomic sequence by a positive hybridization translation assay. Using one of these cloned fragments as a probe for in situ hybridization, 15K transcripts have been localized to a specific region of the male gonad. These studies indicate that the gene for the major sperm protein is regulated by a cell-specific transcriptional control mechanism coincidental with the onset of sexual differentiation in the nematode.
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[
Dev Biol,
1981]
In order to facilitate the biochemical analysis of spermatogenesis in the nematode Caenorhabditis elegans methods have been developed for obtaining large quantities of males and for the isolation of sperm. Males are isolated by a passive filtration method from strains producing high proportions of males and sperm are isolated by physical pressure followed by filtration and differential centrifugation. Biochemical analyses show that sperm contain a major protein component that represents 17% of the total sperm protein. This protein has a molecular weight of 15,600, an isoelectric pH of 8.6, and exists as a dimer. It is shown by immunocytochemical techniques to be a specific product of spermatogenesis. It is localized in the proximal arm of the male gonad and in the sperm of both the male and hermaphrodite but it is not detected in other tissues of the nematode. It is not a nuclear binding protein. Pulse-labeling studies show that this major sperm protein is first synthesized in the proximal arm of the male gonad beginning at 39-42 hr after hatching at 20C. Poly(A) mRNA coding for this protein is first detected in a translatable form just before synthesis of this sperm protein suggesting transcriptional control.
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[
Mol Cell Biol,
1984]
The major sperm protein (MSP) of the nematode Caenorhabditis elegans is a low-molecular-weight (15,000) basic protein implicated in the pseudopodial movement of mature spermatozoa. Its synthesis occurs in a specific region of the gonad and is regulated at the level of transcription (M. Klass and D. Hirsh, Dev. Biol. 84:299-312, 1981; S. Ward and M. Klass, Dev. Biol. 92:203-208, 1982; Klass et al., Dev. Biol. 93:152-164, 1982). A developmentally regulated gene family has been identified that codes for this MSP. Whole genomic blots, as well as analysis of genomic clone banks, indicate that there are between 15 and 25 copies of the MSP gene in the nematode genome. Southern blot analysis also indicates that there is no rearrangement or amplification within the MSP gene family during development. No evidence was found of methylation at various restriction sites surrounding the MSP gene family, and similarly, no correlation between methylation and expression was observed. Three distinct members of this MSP gene family have been cloned, and their nucleotide sequences have been determined. Differential screening of a cDNA clone bank made from polyadenylated mRNA from adult males yielded 45 male-specific clones, 32 of which were clones of MSP genes. One of these cDNA clones was found to contain the entire nucleotide sequence for the MSP, including part of the 5' leader and all of the 3' trailing sequence. Genomic clones bearing copies of the MSP genes have been isolated. At least one of the members of this gene family is a pseudogene.(ABSTRACT TRUNCATED AT 250
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[
J Mol Biol,
1988]
The major sperm proteins (MSPs) are encoded in the Caenorhabditis genome by a multigene family with more than 50 genes dispersed in small clusters at three chromosomal loci. In spite of their dispersed locations, all of the MSP genes appear to be expressed at the same time exclusively in the testis, indicating co-ordinate temporal and spatial regulation of these dispersed genes. Many of the MSP genes must be transcribed, because RNA hybridization with gene-specific probes showed that individual genes each contribute less than 3% to the total poly(A)+ RNA, and 13 out of 14 sequenced cDNAs came from different genes. Primer extension assays from MSP mRNA showed that most of the MSP mRNAs must be initiated at position -35 from the translation start codon. Extensive similarity was found in the first 100 nucleotides of genomic sequence flanking the start codons of ten MSP genes from different chromosomal locations. All MSP genes contained a consensus ribosome binding site, a consensus TATA homology 27 nucleotides distal to the site of mRNA initiation, and ten highly conserved nucleotides adjacent to the site of initiation. All the MSP genes contained the sequence AGATCT located approximately 65 nucleotides upstream from the transcriptional start, but little or no similarity was found more distal to this. Some of these conserved sequences may be cis-acting control elements that ensure the cell and temporal specificity of transcription of these co-ordinately regulated genes.