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  • 1
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 2
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 39-58 
    ISSN: 0192-253X
    Keywords: Drosophila melanogaster ; trisomy 3L ; dosage compensation ; heat shock ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Production of trisomic-3L Drosophila melanogaster has allowed further investigation of compensated levels of gene expression in autosomal trisomies. We find that four enzyme loci on this arm produce diploid levels of gene product in trisomic-3L larvae. For one of these genes, we show that all three alleles are expressed at similar levels. Two genes on 3L display dose-dependent levels of gene product, and their location, relative to the four compensating loci, indicates that these two classes of genes are not regionally separated. In trisomic-2R larvae, the level of enzyme produced from on 2R-linked gene was dose dependent. In contrast, measurements of five loci on the X chromosome in metafemales (X trisomies) suggest that most genes are compensated in these individuals. Heat-shock gene expression in trisomic-3L salivary glands was qualitatively similar to diploids. The quantities of the small hsps (from the 67B cluster on 3L) suggest that these four genes respond independently to the trisomic condition; two produce compensated levels of protein, whereas the other two produce dose-dependent levels of protein. The amount of hsp 83 produced in trisomies was similar to diploids (compensated). However, quantification of hsp 83 RNA showed that a dose-dependent level of transcript was produced. This implies that hsp 83 compensation is controlled post-transcriptionally.
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  • 3
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 93-100 
    ISSN: 0192-253X
    Keywords: heat shock ; phenocopy ; forked ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Heat shock uncovers the recessive forked phenotype when heterozygotes between f36a and wild-type are heated during sensitive periods in pupal development. We call the phenocopy of a mutant in such a heterozygote a heterocopy. The heterocopy in f36a/+ is virtually identical to the mutant phenotype; however, bristles on different parts of the body are affected during different sensitive periods. We discuss the hypothesis that the heat shock acts by affecting expression of the wild-type gene product corresponding to the mutant gene. The sensitive period for heterocopy induction in a specific tissue is proposed to correspond to the normal time of gene expression for the forked gene product in a particular tissue.
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  • 4
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 151-151 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 5
    ISSN: 0192-253X
    Keywords: Dictyostelium discoideum ; revertants of stmF mutants ; cGMP metabolism ; cGMP-specific phosphodiesterase ; suppressor mutations ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: stmF mutants of Dictyostelium discoideum produce long, banded aggregation streams on growth plates and exhibit altered cGMP metabolism. To learn more about the role of cGMP in chemotaxis and the nature of the defect in these mutants, 15 nonstreaming (Stm+) revertants of two stmF mutants were isolated and characterized. Fourteen of the revertants continued to show the elevated cAMP-induced cGMP response and very low cGMP-specific phosphodiesterase (cGPD) activity characteristic of their stmF parents. Parasexual genetic analysis revealed that many of these Stm+ revertants carried phenotypic suppressors unlinked to stmF. One Stm+ revertant, strain HC344, exhibited a low, prolonged cGMP response and relatively high cGPD activity throughout development. To determine whether the elevated cGPD activity in this revertant resulted from increased enzyme production or enhanced enzyme activity, cGPDs were partially purified from the wild-type strain, the stmF parent and revertant HC344, and properties of the enzymes were compared. cGPDs from the stmF mutant and the revertant showed similar differences from the wild-type enzyme in kinetic properties, thermal stability, and sensitivity to certain inhibitors. These results suggest that stmF is the structural gene of the cGPD. In addition, the unusual cGMP response in revertant HC344 appeared to be due to increased production of an altered cGPD.
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  • 6
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 213-238 
    ISSN: 0192-253X
    Keywords: ciliate pattern formation ; expression of mutations ; Tetrahymena thermophila mutations ; reversals of symmetry ; spatial organization of cell surface ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The initial changes of cell-surface organization that occurred as the recessive janAl (janus) mutation of Tetrahymena thermophila first became expressed were elucidated in a special mating scheme in which old macronuclei homozygous for janA+ were synchronously replaced by new macronuclei homozygous for janAl. During this period of onset of expression, the number, regularity, and asymmetry of the ciliary rows remained unchanged. New normal (primary) oral apparatuses (OAs) continued to be formed posterior to old OAs, as in normal cells. At about four fissions after conjugation, abnormal (secondary) OAs with a partial reversal of asymmetry began to appear nearly opposite to the primary OAs, close to but not at the eventual circumferential position of janAl secondary OAs. The array of contractile vacuole pores (CVPs), normally located adjacent to two ciliary rows centered near 22% of the cell circumference to the righ of the primary oral meridian, underwent a two-step transformation: first, the number of adjacent ciliary rows bearing CVPs increased to 3, 4, and sometimes 5, then “skipped” rows appeared within this broadened CVP-arc to split the single set of CVPs into two separated subsets. The CVP transformations occurred gradually and progressively. They began prior to the expression of secondary OAs but accelerated as secondary OAs appeared. As the CVP are became broader, its midpoint shifted somewhat to the right, away from the primary oral meridian, but ended up close to halfway between the primary and secondary oral meridians. The data provide a better fit to an intercalation model than to an alternative double gradient model, suggesting that the janAl mutation alters the large-scale organization of positional values by preventing the expression of a subset of these values and thus provoking reverse-intercalation of the remainder.
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  • 7
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 293-293 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 8
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 297-297 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 9
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 7 (1986) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 10
    ISSN: 0192-253X
    Keywords: Drosophila ; temperature effects ; heat-shock ; cell-lethal mutation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Pulses of various durations at temperatures between 29 and 38°C were applied to developing larvae of Drosophila melanogaster carrying the temperature-sensitive cell-lethal mutation 1 (1)ts726. The results show that it is not possible to reduce the time required for the induction of abnormalities in the mutant by treating larvae with heat pulses at temperatures higher than 29°C. Instead, treatment with high temperature leads to fewer abnormalities than 29°C treatments. Furthermore with high temperature treatments, the mutation has less effect on viability than is seen at 29°C. It is suggested that 1 (1)ts726 leads to abnormalities and death by a temperature-induced imbalance between different physiological or development events, rather than by interfering with the ability of the cell or the organism to withstand high temperature in general.
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  • 11
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 59-74 
    ISSN: 0192-253X
    Keywords: Dictyostelium discoideum ; cell cohesion ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three stage-specific cohesive systems operate in D. discoideum: VEG, elaborated by vegetative cells: AR, by aggregation competent cells; and PAR, by post aggregation stage cells. Previous study of a mutant strain JC-5 had shown the stability of its PAR system (but not the AR) to be temperature sensitive. However, the phenotypic expression of this mutation termed Coh A is complicated by the presence in that strain of a preexisting mutant gene Rde A, which accelerates developmental events generally and alters the pattern of morphogenesis. Genetic evidence presented here indicates that the two mutations have been separated by parasexual recombination yielding a Coh A, Rde A+ segregant class of which strain JC-36 is a prototype.At the permissive temperature, JC-36 follows a morphogenetic sequence like that of the wild type in respect to timing, morphogenetic pattern, and spore appearance. At the restrictive temperature, it forms normal aggregates at the usual time but exhibits two morphogenetic aberrancies during post aggregative development. First, fruit construction is arrested at a stage approximating the 16 hr “Bottle” stage of the wild type, though more squat and blunt tipped, and then the aggregate regresses. Cytodifferentiation into spores and stalk cells is also blocked. Second, a shift of slugs migrating normally at the permissive temperature to the restrictive causes the latter to disintegrate progressively as they leave clumps of cells behind them within the flattened sheath.JC-36 cells developing at the restrictive temperature also exhibited a decrease in EDTA resistant cohesivity attributable on two grounds to the sensitivity of the PAR system. In addition, the disappearance of the AR system completed in the wild type by the Mexicanhat (18-19 hr) stage is indefinitely arrested at an intermediate level in JC-36.
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  • 12
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 113-132 
    ISSN: 0192-253X
    Keywords: eliminated DNA ; facultatively persistent sequences ; macronuclear development ; Tetrahymena thermophila ; phenotypic assortment ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: During conjugation in the ciliated protozoan, Tetrahymena thermophila, a somatic MAC-ronucleus develops from the germinal MICronucleus. Ten to 20 percent of the MIC genome is eliminated during this process. Three repetitive families have been identified which have different levels of repetition in the MIC and are eliminated to different degrees in the MAC. Some members of two of these families persist in the MAC. In this study, we have looked at these persistent sequences in the MAC of cell lines from a variety of sources including several inbed strains, two sets of caryonides, caryonidal subclones, and vegetatively aged cell clones. The results suggest that the sequences that remain in the MAC have a genetic predisposition to persist. However, epigenetic variations occur as the MAC develops so that only some of the persistent sequences are actually observed in a particular MAC. Polymorphisms may be generated if alternative processing of a single MIC segment occurs. These polymorphisms can later be resolved by phenotypic assortment during vegetative growth. These facultatively persistent sequences appear to differ from sequences previously described in this organism.
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  • 13
    ISSN: 0192-253X
    Keywords: ecdysteroid ; prothoracic gland ; temperature sensitive ; Drosophila ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The dominant temperature-sensitive mutation L(3)3DTS (DTS-3) in Drosophila melanogaster causes lethality of heterozygotes during the third larval instar at the restrictive temperature (29°C). Temperature-shift experiments revealed two distinct temperature-sensitive periods, with lethal phases during the third larval instar (which may persist for 4 weeks) and during the late pupal stage. At 29°C mutant imaginal discs are unable to evert in situ, but did evert normally if cultured in the presence of exogenous ecdysterone or when implanted into wild-type larval hosts. The only morphologically abnormal tissue present in the lethal larvae is the ring gland, the prothoracic gland being greatly hypertrophied in third instar DTS-3 larvae. Injection of a single wild-type ring gland rescued these mutant larvae, indicating that the mutant gland is functionally, as well as morphologically, abnormal. Finally, the mutant larvae were shown to have less than 10% of the wild-type ecdysteroid levels. These results are all consistent with a proposed lesion in ecdysteroid hormone production in DTS-3 larvae. A comparison with the phenotypes of other “ecdysone-less” mutants is presented.
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  • 14
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 35-43 
    ISSN: 0192-253X
    Keywords: development ; isozymes ; murine trisomy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We examined developmental changes in the relative activities of three different isozyme systems: aldolase, enolase and phosphoglycerate mutase, in tissues of fetal mice with trisomy 16 and of fetal euploid littermates. We wanted to determine whether morphological abnormalities such as reduced weight and size, which are generally observed in murine trisomy, are reflected at the molecular level. Following electrophoretic separation and subsequent measurement of relative activities of enolase isozymes in brain and phospho-glycerate mutase isozymes in heart, we found no significant differences between trisomy 16 fetuses and their euploid littermates. Synthesis of liver-specific aldolase was, however, delayed in trisomy 16 fetuses.
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  • 15
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 83-89 
    ISSN: 0192-253X
    Keywords: chick blastula ; hypoblast-epiblast interaction ; transcriptional control ; α-amanitin ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Interaction between the epiblast and the primary hypoblast in chick blastula results in induction of the primitive streak (PS) in the epiblast. Alpha-amanitin, a specific inhibitor of poly A-containing RNA synthesis, inhibits formation of the definitive PS. This inhibition is associated with qualitative changes in the pattern of protein synthesis in the hypoblast but not in the epiblast. The protein pattern of the component areas of the epiblast shows increase in some polypeptides after treatment with α-amanitin. By contrast, α-amanitin resulted in a decrease in synthesis of several polypeptides, which are either undetectable or weakly present in the hypoblast. The α-amanitin-sensitive translational products of the embryonic genome that are observed in the hypoblast may have specific functions in the control of PS induction and stabilization.
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  • 16
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 121-122 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 17
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 99-119 
    ISSN: 0192-253X
    Keywords: Drosophila ; tissue polarity ; frizzled ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The epidermis of Drosophila has a tissue polarity that is manifested by a parallel array of polarized structures (primarily hairs and bristles). The production of normal tissue polarity requires the function of the frizzled (fz) locus. We have isolated a large number of alleles at this locus and have phenotypically characterized more than 25 of them. We have found extensive allelic variation that a previous study failed to detect. Most of the alleles fall into a hypomorphic to amorphic series. Two alleles, however, have unusual properties. These alleles, which in general are moderately strong alleles, fail to produce a rough eye phenotype that is characteristic of all the other moderate or strong fz alleles. Thus, these two alleles are tissue specific in effect. Furthermore, these two alleles also have a neomorphic or antimorphic effect on hair polarity in one region of the wing.
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  • 18
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 165-177 
    ISSN: 0192-253X
    Keywords: embryonic antigen ; tumor mutants ; oncodevelopmental molecule ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The 63-kDa antigen recognized by the monoclonal antibody F7D6 is present in all Drosophila embryonic cells and disappears from most tissues as each one reaches its final, differentiated state. Larval tissues lose the antigen around the time of hatching, imaginal tissues lose it during metamorphosis, and germ cells lose it during gametogenesis (Bedian et al: Devel Biol 115:105-118, 1986). The nervous system and spontaneously contracting musculature of the gut and gonads are exceptions and remain antigen positive at all stages. The F7D6 antigen appears to be associated with dividing, undifferentiated cells and electrogenic cells. This prompted us to test tumors for antigen presence. We tested four different recessive mutants that give rise to four different types of tumorous transformation: the embryonic tumor Notch, several larval melanotic tumors, the imaginal disc tumor 1(2)gl, and three alleles of the ovarian tumor otu. In all cases, tumorous tissues in homozygotes contained the F7D6 antigen. The electrophoretic mobility of the antigen appeared to be unaltered in tumorous tissues compared to normal cells, but the antigen is expressed at higher levels. The antigen is found on the cytoplasmic surface of plasma membranes and appears to be a marker of undifferentiated normal and tumorous cells. Similarities and differences between the F7D6 antigen and Drosophila c-src protein are discussed.
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  • 19
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. i 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 20
    ISSN: 0192-253X
    Keywords: microinjection ; familial amyloidotic polyneuropathy ; fertilized egg ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: To analyze the regulation of transthyretin gene expression we have produced transgenic mice by microinjecting cloned human transthyretin genes into fertilized eggs of C57BL/6 mice. The 7.6-kilobase (kb) human transthyretin gene containing about 500 base pairs (bp) in the upstream region was used for microinjection. Seven out of nine transgenic mice had detectable amounts of human transthyretin in serum when analyzed by enzyme-linked immunosorbent assay.Transthyretin mRNA was detected in liver and yolk sac but not in other tissues including brain. The amount of mRNA was variable among transgenic mice and was about one-tenth of mouse endogenous transthyretin mRNA. Human and mouse transthyretin mRNAs were detected in liver of fetus and yolk sac at 13 days of gestation and unlike yolk sac the level of mRNA in liver increased gradually during development and reached the maximum at around 17 days of gestation. Human transthyretin was associated with mouse transthyretin to form tetramers as judged from the dilution curve of enzyme-linked immu-nosorbent assay and the spur formation in Ouchterlony assay.
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  • 21
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 281-293 
    ISSN: 0192-253X
    Keywords: mouse ; human ; cow ; maps ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Technological advances in the 1970s encouraged the mapping of homologous gene loci in different mammalian species, including mouse and man. One hundred eighty-five homologous loci have now been mapped in these two species. Conservation of linkage is sufficient to identify substantial segments of the two genomes that have been left intact since their divergence from a common ancestor. The recognition of these conserved segments allows experimental manipulation of mouse chromosomes or chromosomal regions to produce models of human chromosomal anomalies of medical importance.Comparative gene mapping has been extended beyond mouse and man and the genomes of some species, including domestic cattle, appear to be more highly conserved relative to humans than the mouse. Such species may be particularly useful in providing models of human chromosomal anomalies that cannot be duplicated in laboratory mice.
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  • 22
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 9 (1988), S. 23-35 
    ISSN: 0192-253X
    Keywords: maternal-specific transcripts ; genomic clones ; developmental RNA expression ; in situ chromosomal localization ; embryogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The unique cellular and genetic events which occur during the first few hours of Drosophila embryogenesis suggest that there are genes whose function is entirely or largely limited to this stage; this is supported by both genetic and molecular evidence. To identify some of these genes and characterize the relative contribution of specifically maternal and specifically zygotic transcription to early embryogenesis, we used competition and differential screening of a Drosophila genomic DNA library to obtain blastoderm- and maternal-differential sequences [Roark et al.: Dev Biol 109:476-488, 1985]. We describe here the Eco RI restriction fragments, chromosomal location, and size and developmental pattern of expression of the RNAs transcribed from 19 maternal-differential sequences. Five sequences encode maternal-specific transcripts (50-150-fold more abundant in maternal RNA than at any other stage). The maternal-specific and maternal-differential sequences are located at single sites on all major chromosome arms. Comparison of these sites with the sites of presently mapped maternal-effect genes shows several possible correlations, including one region containing three maternal-effect lethal mutations and two maternalspecific sequences.
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  • 23
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 9 (1988), S. 69-69 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 24
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 9 (1988) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 25
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    Developmental Genetics 8 (1987), S. 91-98 
    ISSN: 0192-253X
    Keywords: wing size ; miniature ; cell size ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: To elucidate the mechanisms whereby genes and environment influence wing size, we investigated the effects of various rearing temperatures and larval crowding conditions on the wings of the mutant miniature and wild-type fruit flies. In adults we monitored wing size, cell number, wing thickness, cell density; in larval imaginal discs we looked for cell death. Cell density was inversely proportional to wing size. Of particular interest was the finding that smaller wings tend to be thicker. Electron microscope studies showed that the miniature wing layers are grossly abnormal. We hypothesize that these abnormalities are due to abnormal cell flattening of the wing epithelial cells, and we conclude that gene and environmental effects on cell flattening may be an important component in determining cell density and hence organ size.
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  • 26
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    Developmental Genetics 8 (1987), S. 123-123 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 27
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 125-133 
    ISSN: 0192-253X
    Keywords: retrovirus ; embryonal carcinoma ; embryonic gene ; DNA methylation ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Northern blot analysis and in vitro nuclear transcription assays were performed in order to clarify conflicting reports on the expression of intracisternal A particle (IAP) genes in embryonal carcinoma (EC) cell lines. Results demonstrate that post-transcriptional mechanisms control the final steady-state levels of IAP RNA in EC cells. IAP genes were further found to be undermethylated in IAP-expressing EC cell lines.
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  • 28
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    Developmental Genetics 8 (1987), S. 187-187 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 29
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 189-194 
    ISSN: 0192-253X
    Keywords: H-Y antigen ; skin grafts ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The influence of the major histocompatibility complex (MHC) on the survival of H-Y-incompatible skin grafts in rats has been determined by challenging normal and previously sensitized females of various isogenic and congenic strains with male trunk or ear skin isografts. The MHC's influence on the potency of H-Y has also been evaluated by determining the survival of male parental strain ear skin grafts on sensitized (with F1 hybrid male cells) F1 hybrid females of two different MHC congenic strains. The results indicate that, as in mice, the MHC has a dual affect on H-Y; it is involved in determining the ability of females to respond to the antigen as well as influencing its potency.
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  • 30
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 233-247 
    ISSN: 0192-253X
    Keywords: DNA dispersion ; human β-globin ; reverse transcription ; evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A human bacteriophage clone containing adult β-globin genes with four Alu sequences was microinjected to produce transgenic mice. Southern blot analysis on the spleen of a transgenic mouse revealed an unusual hybridization pattern that suggested extensive dispersion of human DNA throughout the mouse genome. This pattern was reproducible using several restriction enzymes, including a noncutting enzyme. The hybridization pattern was not observed in other tissues, and sequences were not detected in progeny using the bacteriophage probe. However, hybridization of spleen DNA of offspring against a human Alu probe revealed genetic transmission of human Alu sequences. The results suggest dispersion of microinjected Alu sequences throughout the genome.
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    Developmental Genetics 8 (1987), S. 321-337 
    ISSN: 0192-253X
    Keywords: T-DNA ; T-cyt gene ; plant promoter structure ; plant development ; plant gene regulation ; plant defense-related mRNAs ; Agrobacterium tumefaciens ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 32
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    Developmental Genetics 8 (1987), S. 375-387 
    ISSN: 0192-253X
    Keywords: urease ; isozymes ; clones ; null mutants ; soybean ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The soybean (Glycine max [L.] Merr.) contains two urease isozymes whose expression is regulated in a tissue-specific and temporal manner. The ubiquitous urease is expressed in all tissues examined (leaf, embryo, seed coat, cell culture); the embryo-specific urease is synthesized exclusively in the developing embryo. The embryo-specific urease accumulates during seed development while the ubiquitous urease is found in highest levels during early development of both leaves and seeds. We have isolated mutants which fall in three phenotypic classes lacking one or both urease isozyme activities. Genetic analysis has thus far identified three unlinked loci which control the expression of urease(s). Genomic and cDNA clones of urease structural genes have also been recovered and we are working to assign these to genetic loci by sequence and RFLP analyses. That the ubiquitous urease isozyme is expressed in cell culture makes it possible to include cell culture in physiological and developmental studies. Additionally, we have developed direct selections for urease-negative mutants, and their revertants, in cell culture. These selections will facilitate the study of the expression of cloned urease genes in genetically transformed tissue.
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  • 33
    ISSN: 0192-253X
    Keywords: tubulin genes ; microtubules ; Arabidopsis thaliana ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Microtubules are important components of the cytoskeleton of plant cells and play key roles in plant growth and morphogenesis. Recent molecular studies have begun to elucidate the structure and expression of plant genes coding for the major components of microtubules, α- and β-tubulin. Tubulin amino acid sequences deduced from the DNA sequences of eight higher plant tubulin genes are 79-87% homologous with constitutively expressed mammalian tubulins. The genome of the model plant system Arabidopsis thaliana contains four dispersed α-tubulin sequences and at least seven β-tubulin sequences, only two of which appear to be linked. Of the five A. thaliana genes whose expression has been analyzed, the transcripts of one α-tubulin and one β-tubulin gene are constitutively expressed in roots, leaves, and flowers. A second α-tubulin gene is expressed predominately in flowers; the transcripts of the second and third β-tubulin genes are found predominately in leaves or in roots, respectively.
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    Developmental Genetics 9 (1988), S. 1-12 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 35
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    Developmental Genetics 9 (1988), S. 37-48 
    ISSN: 0192-253X
    Keywords: eggshell ; female sterile mutant ; endochorion ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Four female-sterile mutants, fs(l)K451, fs(1)K1214, fs(1)K575TS, and fs(1)384, were studied in terms of chorion structure and chorion protein composition. The first three of these mutants cause morphological defects, ie, substantial underproduction and disruption of the endochorion, correlated with underproduction of the six major chorion proteins, s15-s38; the phenotypes are consistent with the observation that these mutants interfere with amplification of the major chorion genes that encode the s15-s38 proteins [Orr et al.: Proc Natl Acad Sci USA 81:3773-3777, 1984; Komitopoulou et al.: Dev Genet 7:75-80, 1986]. The fourth mutant, fs(1)384, and its alleles do not interfere with production of the major chorion proteins and the morphologically detectable bulk of the endochorion but lead to failure of endochorionic organization. Apparently this complementation group is responsible for a minor chorion product, which is evidently important morphogenetically and which is processed posttranslationally in a complex manner [Bauer and Waring: Dev Biol 121:349-358, 1987].
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    Developmental Genetics 9 (1988), S. 91-120 
    ISSN: 0192-253X
    Keywords: actin filament bundles ; ethyl methane sulfonate ; female sterile mutants ; in terallelic complementation ; oocyte determination ; ovarian tumor genes ; phenocritical thresholds ; polyfusomes ; polytene nurse cell chromosomes ; polytrophic meroistic ovaries ; pseudonurse cells ; Q-T-P-O values ; rhodaminyl phalloidin ; temperature-sensitive mutants ; transformed oocytes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ovarian tumor gene behaves as if it encodes a product (OGP), which is required durirng several early steps in the transformation of oogonia into functional oocytes. Seventeen ethyl methane sulfonate-induced mutations have been studied, and their mutant phenotypes can be explained as graded responses by individual germ cells to different levels of OGP synthesized by the mutant germ cells themselves. The lowest and highest levels of OGP appear to be produced by otu10 and otu14, respectively. The 15 mutants with intermediate OGP levels are temperature sensitive; subnormal temperatures improve ovarian development, while above-normal temperatures suppress it. A subgroup ofthese mutants are unable to form a system of actin microfilament bundles in the cortical cytoplasm of their nurse cells during stage 10B, and these defective nurse cells are unable to transport their cytoplasm to the oocyte, as normally happens between stages 10B and 12. In addition to its role in the actin-mediated transport of nurse cell cytoplasm, OGP also appears to alter the morphology of giant polytene chromosomes, which form as the nurse cells undergo endocycles of DNA replication. Genetic evidence suggests that otu also encodes a second product (SP) that is utilized late in oogenesis. SP is required for the synthesis in the ooplasm of glycogen-rich, beta yolk spheres. Products of the otu gene also play a vital but unknown role in embryogenesis.
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    Developmental Genetics 9 (1988), S. 181-191 
    ISSN: 0192-253X
    Keywords: cell interactions ; hematopoiesis ; mutagenesis ; SV40 ; fetal liver ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Several lines of transgenic mice were produced by pronuclear injection of a full-length cDNA encoding a mutant dihydrofolate reductase (DHFR, E.C. 1.5.1.3). The mutation causes altered enzyme kinetics for folate reduction as well as low affinity for methotrexate (MTX). One line of mice carrying the plasmid displays a moderate-to-severe anemia that is evident in fetuses and newborn mice and that moderates with age. RNA studies revealed high levels of transcription of the mutant gene in the fetal and adult liver, and low or absent expression in adult bone marrow. Transcription of the mutant gene was not found in the fetal liver of other pedigrees examined. The data thus suggest that expression of this mutant gene in the main hematopoietic organ of the fetus adversely affects erythropoiesis by altering the cellular environment for erythroid differentiation, and that translocation of the site of hematopoiesis to bone marrow, where the foreign gene is not expressed, leads to normalization of red cell production.
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    Developmental Genetics 9 (1988), S. 213-213 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 39
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    Developmental Genetics 9 (1988), S. 683-698 
    ISSN: 0192-253X
    Keywords: variegation ; bristles ; pigmentation ; patterns ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In D. hydei two new mutants, In(1)f3 and IN(5)Z, show obvious mosaic gene expression. Their phenotypic expression is susceptible to the breeding temperature and to the addition of a supernumerary Y chromosome to the chromosome set. In this respect the mutants resemble standard cases of position-effect variegation based on the action of heterochromatin. However, since neither centromeric nor sex chromosomal heterochromatin apparently are involved, the mutations point to a new type of variegation provoked by euchromatic sections. The mosaic patterns of these mutants, in particular those of In(1)f3, will be described.
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    Developmental Genetics 6 (1985), S. 1-25 
    ISSN: 0192-253X
    Keywords: developmental mutants ; axolotl ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Mexican axolotl (Ambystoma mexicanum) has enjoyed wide use in experimental embryology for over 100 yr. Its usefulness has been extended into the area of developmental genetics largely due to the contributions of R. Briggs and R. R. Humphrey at Indiana University. To date over 30 mutants have been described, almost all of which affect development. Some of these have been discovered in inbred strains while others have been uncovered in recent Mexican imports. These mutants can be subdivided into several major classes. Maternal effect mutations lead to deficiencies in informational, structural, or metabolic components of the egg essential to early development prior to the time at which the embryo's own genome becomes active. In contrast, the developmental lethals affect later stages in embryogenesis when both morphogenetic and biochemical events are determined exclusively by the genotype of the embryo. Most lead to death at about feeding stage. Some, the cell lethals, are believed to suffer from fundamental metabolic defects affecting all parts of the embryo. Others affect the development of specific organs or tissues. The developmental nonlethals also affect specific systems, but ones that are not essential to survival. Some affect the development and survival of pigment cells and these, along with isozyme variants, are useful as markers in developmental experiments.A number of the mutants have been studied in detail, but others scarcely at all. The purpose of this review is to bring them to the attention of all developmental biologists in the hope that their potential will be even more widely recognized.
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    Developmental Genetics 6 (1985) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 42
    ISSN: 0192-253X
    Keywords: Tripsacum dactyloides ; Zea mays ; tripsacoid maize ; abnormal development ; ribosomal DNA ; restriction site change ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Some of the derivatives of a cross of maize (Zea mays L.) × Tripsacum dactyloides (L) L (2n = 72) have abnormal development leading to strange and striking morphologies. The Tripsacum chromosomes in these “tripsacoid” maize plants (with Tripsacum-like characteristics) were eliminated and the maize chromosomes were recovered through repeated backcrossing to maize. As an initial attempt to analyze the DNA alterations in tripsacoid maize, we have detected a few restriction site changes in the ribosomal DNA repeat of these plants (Hpa II, Bal I, Sst I, Mbo II, and Sph I) and a new Sph I site was mapped to the spacer region between the 26S and 17S genes. Several possible mechanisms for the generation of a new restriction site are discussed, and we propose that the transient presence of Tripsacum genome during the backcrossing in some way induced a rapid amplification and fixation of new restriction sites in a relatively short period of time.
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    Developmental Genetics 6 (1985), S. 133-150 
    ISSN: 0192-253X
    Keywords: vitellogenesis ; Drosophila melanogaster ; egg shell ; oogenesis ; vitellogenin ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ovarian follicle cells of wild type Drosophila melanogaster simultaneously secrete yolk polypeptides (YP1, YP2 and YP3) and vitelline membrane proteins. In order to understand the relationship between these two secretory activities, we have investigated the ultrastructure of a female sterile mutation that alters YP1 secretion and vitelline membrane deposition. Homozygous fs(1)1163 females lay eggs that collapse and contain reduced quantities of YP1. Secretory granules in follicle cells contain an electron-translucent component that is assembled into the developing vitelline membrane in both mutant and wild-type ovaries, and an electron-dense component that disperses after secretion in wild-type ovaries. Mutant ovaries differ from wild-type by (1) having larger secretory granules (2) forming clumps of the dense secretory component within the developing vitelline membrane (3) accumulating more tubules in the cortical ooplasm of vitellogenic oocytes, and (4) possessing altered yolk spheres. Mutant ovaries implanted into wild-type hosts showed no improvement in the secretory granules and slight improvement in the vitelline membrane clumps but amelioration of the oocyte phenotypes. Since genetic evidence suggests that the fs(1)1163 mutation resides in or near the Yp1 gene and biochemical data show that the mutation alters YP1 structure, we conclude that the ultrastructural phenotypes are due to a structurally abnormal YP1 in the mutant. The alteration in vitelline membrane structure caused by the dense clumps could account for collapsed eggs and, hence, the female sterility of the mutant.
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    Developmental Genetics 6 (1985) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 45
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    Developmental Genetics 6 (1985), S. 247-255 
    ISSN: 0192-253X
    Keywords: Drosophila ; triploid intersexes ; sex differentiation ; dosage compensation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Triploid intersexes homozygous for a mutant (msl-2) known to impede the hyperactivation of the X chromosome in diploid males differentiate into adults, sexually indistinguishable from their heterozygous sibs. A shift toward female sexual differentiation mediated by manipulating the rearing temperature is accompanied by an apparent increase in the level of an X-linked gene product. This unexpected result is rationalized in terms of differential lethality of individuals at the two extremities of the distribution of X-activity levels in intersexes raised at a particular temperature. No evidence of a mosaicism comparable to the sexual mosaicism exhibited could be found with respect to an X-linked gene product in triploid intersexes.
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  • 46
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    Developmental Genetics 6 (1985), S. 281-291 
    ISSN: 0192-253X
    Keywords: DNA insertion ; reversion ; variegation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A spontaneous white mutation recovered in Drosophila mauritiana is unstable and reverts to normal eye color at a frequency greater than 4 per 1,000 ×-chromosomes. Germ line reversion occurs at a high rate in D. mauritiana males and in interspecific hybrid females, while the rate is depressed in D. mauritiana females. These events are not restricted to the germ line, as cases of variegated patterns of eye pigmentation, indicating somatic reversion, are recovered at a frequency comparable to that of the male germ line reversion rate. Germ line reversion events are genetically stable, while the somatic variegation patterns are not heritable. The patterns of eye pigment variegation produced suggests that reversion events are occurring throughout development. Whole genome DNA digests blotted and probed with the cloned D. melanogaster white gene indicate that this unstable white mutation in D. mauritiana is associated with an insertion of DNA that is lost upon reversion to wild type, indicating that this DNA insert is in fact a transposable element.
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    Developmental Genetics 7 (1986), S. 21-34 
    ISSN: 0192-253X
    Keywords: Drosophila ; pattern formation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The distribution of the soluble form of NADP+ -dependent isocitrate dehydrogenase (ICDH) was examined in Drosophila melanogaster imaginal discs. Development of the enzyme patterns and the specific transformations of the patterns by homoeotic mutants were studied. ICDH pattern formation was followed in eye-antennal discs and wing discs from the late 2nd instar stage through 3rd instar and 8 hours into prepupal development. The patterns formed gradually in both disc types. The most interesting pattern developed in the eye portion of the eye-antennal disc complex. ICDH distribution as well as staining intensity correlated well with differentiation of the ommatidia. The spatial distribution of ICDH within the discs was under genetic control. The patterns reflected the state of determination of the disc. When the presumptive tissue type was transformed via mutant homoeotic genes to different determinative states, the ICDH pattern likewise transformed to the pattern characteristic of the newly acquired structure.
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    Developmental Genetics 7 (1986), S. 35-49 
    ISSN: 0192-253X
    Keywords: zea mays ; embryogenesis ; defective kernel mutants ; genetic regulation ; maize ; leaf primordia ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The defective kernel (dek) mutants of maize are altered in both their embryo and endosperm development. Earlier studies have indicated that some of the dek mutants are unable to form shoot apical meristems or leaf primoirda. We have examined three embryo lethal dek mutants of this type, ptd*-1130, cp*-1418, and bno*-747B, to obtain a developmental profile for each. Allelism tests show that these three mutants are not allelic. Embryos were examined in early, mid-, and late kernel development as well as at kernel maturity by dissection and sectioning procedures and also at kernel maturity by scanning electron microscopy. All three mutants lag behind normal embryos in their rate of development. Embryos of ptd*-1130 reached the transition stage by early kernel development and progressed no further but underwent cell enlargement and necrosis during late kernel development. Embryos of cp*-1418 reached an early coleoptilar stage by midkernel development. They subsequently increased in size but did not form any leaf primordia. At kernel maturity, they no longer had a shoot apical meristem but often had a well formed root meristem. They appeared to remain healthy and did not become necrotic. Embryos of bno*747B reached the early coleoptilar stage by early kernel development but progressed no further. By kernel maturity, they had grown into masses of irregularly shaped embryonic tissue that no longer resembled any normal embryo stage but were not necrotic. None of these three mutants responded to attempts to support continued embryo development when cultured, but all three mutants formed callus on N6 and MS media supplemented with 2,4-D. These results indicate that these mutants are all uniformly blocked at specific stages early in embryonic development, have different subsequent developmental fates, and represent three different genes performing unique functions that are essential for embryogenesis.
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    Developmental Genetics 7 (1986), S. 81-97 
    ISSN: 0192-253X
    Keywords: Hawaiian ; Drosophila grimshawi ; yolk proteins ; ovarian follicle development ; fat body ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In analyzing the in vitro pattern of protein synthesis by the fat body and ovaries of the Hawaiian species Drosophila grimshawi, we have found that the ovaries synthesize much more protein than the female fat body and that the majority of the synthesized proteins are retained by the ovarian tissues. In contrast, the fat body secretes most of the proteins into the culture medium. Vitellogenins are the major class of proteins synthesized and released into the medium by both tissues. The synthesis of the three vitellogenin proteins (V1, V2, V3,) is noncoordinate in the two tissues. Ovaries synthesize much more of the V2 protein, less V1 and very little V3, whereas fat body synthesies more Vs1 protein with lesser qu antities of the other two. The follicle cells were identified as the site of ovarian vitellogenin synthesis in D. grimshawi, confirming the findings in D. melanogaster. In D. grimshawi, the three vitellogenins are synthesized by the follicle cells in a noncoordinate and developmentally regulated manner. V2 and V1 are the predominant proteins at the onset of vitellogenesis (S8-9); their production peaks together with that of V3 a few hours later (S10) and then decreases to quantitiesequalto that of V3 during early choriogenesis (S11). During active choriogenesis (S12), V2 and V1 cease to be synthesized, but V3 synthesis continues. The vitellogenins synthesized by the follicles in vitro are released into the medium and not incorporated into the oocyte.
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    Developmental Genetics 7 (1986) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 51
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    Developmental Genetics 7 (1986), S. 177-185 
    ISSN: 0192-253X
    Keywords: sex reversal ; X-chromosome inactivation ; testis formation ; sex determination ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mice heterozygous for the T(X;16)16H translocation and carrying Sxr on their normal (inactive) X chromosome (ie, T16H/X Sxr individuals) may develop as males, females, or hermaphrodites. The proportion of males varied from 22% to 65% depending on the source of the normal X chromosome. A model is proposed, according to which relatively small variations in the spreading of inactivation from the X chromosome into the attached Sxr fragment produce large changes in the proportion of males. Testis weight in T16H/X Sxr males was found to be significantly smaller than in X/X Sxr males, irrespective of the source of the normal X chromosome.
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    Developmental Genetics 7 (1986), S. 205-221 
    ISSN: 0192-253X
    Keywords: gene dosage effects ; Drosophila sex determination ; maternal-zygotic gene interactions ; tandem duplications ; maternal effects ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The daughterless (da) gene provides an essential maternally supplied component for Drosophila sex determination and dosage compensation. In this connection, it is required as a positive regulator of a female-specific master regulatory gene, Sex-lethal (Sxl). In addition, zygotic da gene function is required for male and female viability. Thus, the phenotype da is complex; it includes both maternal and zygotic aspects, as well as both sex-specific and nonsex-specific aspects. Assessment of wild-type da function has relied on the characterization of only a single leaky mutant da allele. In order to better understand the nature of this allele and the relationships between the various aspects of its complex phenotype, tandem duplications of both the mutant and wild-type da alleles were isolated and used in a dose study of this gene's function. Three conclusions were reached: (1) by the most stringent genetic criteria, the mutant da allele is a simple hypomorph, an allele with reduced but non-zero levels of wild-type functions; (2) since increased dose of da+ had no effect on viability or progeny sex ratio, this gene seems not to be a dose-sensitive element of the X/A ratio sex determination signal; and (3) expression of the maternal da+ allele does make a contribution to the nonsex-specific developmental processes that require zygotic da+ function; however, that contribution is clearly minor. In contrast, the zygotic genotype with respect to da appears to have no effect on the expression of Sxl+ in the zygote, the sex-specific process that requires maternal da+ function.
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    Developmental Genetics 8 (1987), S. 27-34 
    ISSN: 0192-253X
    Keywords: W locus ; mouse ; chromosome 5 lethal ; implantation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A recessive lethal mutant linked to Wsh causes the death of homozygous embryos between 4.5 and 5.5 days postcoitum (pc). Histological examination of implantation sites from intercross and backcross matings indicates that homozygotes are not all evident at 4.5 days pc, when embryos have begun to form trophectoderm giant cells and primitive endoderm, but are degenerating by 5.5 days pc, with only a few primary giant cells remaining after this time. The mutants thus form blastocysts that initiate the implantation process but the inner cell mass and polar trophectoderm fail to develop further. In vitro examination and culture of blastocysts indicated that the mutant homozygotes hatch from the zona pellucida and outgrow, although they do so somewhat more slowly than normal embryos. After 3 days of culture, the inner cell masses of mutant outgrowths may be smaller than normal. Since the gene has no known heterozygous effect and the primary gene function remains unknown, the mutant has been given the provisional symbol l(5)-1 for the first lethal on chromosome 5.
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    Developmental Genetics 8 (1987), S. 45-58 
    ISSN: 0192-253X
    Keywords: white-mottled ; Malpighian tubules ; gene action ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Riboflavin deposition in organs of Drosophila hydei was studied by means of a growth test using a riboflavin-deficient strain of the fungus Aspergillus nidulans. In wild-type animals, riboflavin is deposited in Malpighian tubules (MT) and testes but not in adult eyes. Certain white (w) mutants do not contain riboflavin, whereas intermediately colored w mutants contain minor amounts of the substance. Riboflavin-containing MT cells contain special globules that can be fixed and stained with the redox dye phenazine-methosulphate. The number and size of these granules is related to growth effect and point to a role of the w locus in the intracellular deposition of riboflavin in special organs. In white-mottled (wm) position-effect variegation mutants, a significant correlation was found between the extent of variegation (percentage of yellow cells) and riboflavin content (growth effect) of the MT. However, the individual variation of cell phenotype was extremely large and exaggerated types were observed indicating “overdominance” of the rearranged w+ gene. This contradicts an unsubstantiated dogma of position-effect variegation that assumes that the affected gene simply switches between the on and off state, as is discussed.
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    Developmental Genetics 8 (1987), S. 73-82 
    ISSN: 0192-253X
    Keywords: isoelectric focusing ; corticosterone ; gene assignment ; alanine transferase ; tyrosine aminotransferase ; liver cytosol ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The amount of cytosolic glucocorticoid receptor in liver of Ts18, Tsl6, and Tsl9 vs euploid mouse fetuses was studied after incubation of [3H]dexamethasone with cytosol followed by isoelectric focusing on polyacrylamide gels. In addition, corticosterone concentrations and enzyme activities of alanine aminotransferase and tyrosine aminotransferase were measured in the cytosol of the livers. The amount of glucocorticoid receptor in the cytosol fractions of the livers was always higher in the Tsl8 than in the euploid fetuses of the same litter. It was also significantly (P 〈 0.0005) higher if pooled data from different litters were analyzed. The ratio of the glucocorticoid receptor in Ts l8 vs euploid mice varied between 1.3 and 4.7, with a mean of 2.1. In contrast, the glucocorticoid receptor levels in Tsl6 and Tsl9 fetuses were not different from the corresponding euploid controls. Comparing the corticosterone levels of the three trisomies tested with the corresponding euploid fetuses, no significant differences were found, indicating that the markedly elevated cytosolic glucocorticoid receptor concentrations in Tsl8 were not due to different corticosterone levels. This finding is consistent with the assignment of the glucocorticoid receptor gene to chromosome 18 in the mouse. There was no correlation betwen glucocorticoid receptor levels and the activity of the two glucocorticoid inducible enzymes tested in the liver of mouse fetuses.
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  • 56
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    Developmental Genetics 8 (1987) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 57
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    Developmental Genetics 8 (1987), S. 135-150 
    ISSN: 0192-253X
    Keywords: mouse ; NADP-isocitrate dehydrogenase ; electrophoresis ; gene regulation ; allele-specific expression ; heart ; kidney ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The murine “housekeeping” enzyme, cytosolic NADP-isocitrate dehydrogenase (E.C. 1.1.1.42) (genetic locus:Idh-1), exhibited a complex pattern of allele-specific expression. Protein electrophoresis on cellulose-acetate gels and determination of relative enzymatic activity by means of densitometry revealed that in heart tissue (but not liver tissue) of certain hybrid crosses the AA-homodimer was underrepresented relative to total enzymatic activity, and the degree of underrepresentation changed during development. In mixtures of homozygous tissue extracts of heart tissue (but not liver tissue) the AA-homodimer was underrepresented relative to the BB-homodimer. Relative activity of allelic isozymes varied as a function of tissue (heart versus liver), age, and the parental source of the Idh-1a allele, but did not vary as a function of sex.Allele-specific expression was also exhibited in kidney tissue of the same animals. In adult male kidney tissue extracts from heterozygotes, the AA-hornodimer was underrepresented relative to total enzymatic activity; in adult female kidney tissue extracts from heterozygotes, a more codominant phenotype was observed. Tissue extracts from immature hybrid animals exhibited a phenotype midway between the adult male and adult female phenotypes. Tissue extracts from castrated males exhibited a phenotype equivalent to that seen in females. Relative activity of allelic isozymes in kidney varied as a function of age and sex, but did not vary as a function of the parental source of the Idh-1a allele.While cytosolic NADP-IDH is a “housekeeping” enzyme, expressed in multiple tissues of the mouse, differences in the relative intensities of allelic isozyme bands provide evidence for tissue- and stage-specific regulatory variation.
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    Developmental Genetics 8 (1987), S. 179-185 
    ISSN: 0192-253X
    Keywords: differentiation ; melanogenesis ; tyrosinase ; albino ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Albino mutation in animals blocks pigmentation owing to a deficiency in tyrosinase, although it does not affect the differentiation of colorless melanocytes from the neural crest. In the albino Japanese quail (al, sex-linked), it was demonstrated that morphologically normal melanocytes differentiated from neural crest cells in culture and that these cells contained unmelanized melanosomes as expected for the mutant cells. The mutant melanocytes, however, were shown to exhibit tyrosinase activity in the Golgi-endoplasmic reticulum-lysosome region and in the Golgi vesicles. Our results seem to indicate that the mutation at the al locus affects the transport of tyrosinase from the Golgi area to melanosomes.
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  • 59
    ISSN: 0192-253X
    Keywords: lactate dehydrogenase ; spermatogenesis ; multigene enzyme family ; somatic cell hybrids ; gene mapping ; evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: From the data presented in this report, the human LDHC gene locus is assigned to chromosome 11. Three genes determine lactate dehydrogenase (LDH) in man. LDHA and LDHB are expressed in most somatic tissues, while expression of LDHC is confined to the germinal epithelium of the testes. A human LDHC cDNA clone was used as a probe to analyze genomic DNA from rodent/human somatic cell hybrids. The pattern of bands with LDHC hybridization is easily distinguished from the pattern detected by LDHA hybridization, and the LDHC probe is specific for testis mRNA.The structural gene LDHA has been previously assigned to human chromosome 11, while LDHB maps to chromosome 12. Studies of pigeon LDH have shown tight linkage between LDHB and LDHC leading to the expectation that these genes would be syntenic in man. However, the data presented in this paper show conclusively that LDHC is syntenic with LDHA on human chromosome 11.The terminology for LDH genes LDHA, LDHB, and LDHC is equivalent to Ldhl, Ldh2, and Ldh3, respectively.
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    Developmental Genetics 8 (1987), S. 295-304 
    ISSN: 0192-253X
    Keywords: sequence ; cDNA ; fetal pig ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A cDNA clone of porcine alpha1 acid glycoprotein (α1AGP) has been isolated and sequenced. Sequence homologies between porcine, human, and rat indicate that porcine α1AGP is similar in structure to the rat and human proteins. RNA blots from days 40, 60, 80, and 110 fetal, newborn, and adult livers showed that α1AGP mRNA is relatively abundant throughout fetal development, particularly at the later stages and in the newborn; there is a rapid decline in abundance following birth. From birth to 3 days of age, there is a three- to four-fold decline in abundance, and α1AGP mRNA is approximately 100 times less abundant in the adult liver than in that of perinatal pigs. Southern blots showed that α1AGP is probably a single-copy gene. The isolation of a cloned cDNA for porcine α1AGP provides a tool to investigate the molecular mechanisms involved in the developmental regulation of the gene and to correlate changes in gene expression during development with fetal growth and well being.
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  • 61
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    Keywords: mental retardation ; Down syndrome ; cholinergic neurons ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this study, we examined the neurochemical profiles of selected brain regions (cerebral hemispheres, diencephalon/brainstem) in fetal (day 14 to 18 gestation) trisomy 19 (Ts19) mice. The neurochemical characteristics we observed in Ts19 mice were quite different from those we observed previously in Ts16 mice. Choline acetyltransferase (ChAT) activity was reduced significantly in the cerebral hemispheres, but not in the brainstem/diencephalon, of the fetal Ts19 mouse brain, suggesting a selective vulnerability of telencephalic cholinergic neurons. Additionally, the activity of glutamic acid decarboxylase (GAD) was reduced significantly in both hemispheres and diencephalon/brainstem of late gestation Ts19 fetuses, suggesting a selective vulnerability of GABAergic neurons as well. While the levels of catecholaminergic and dopaminergic markers were reduced significantly at late gestational ages, the relative rate of turnover of dopamine (DA), measured by the ratio of DOPAC/DA, was elevated significantly in Ts19 mice. Neither reduction in the thickness of various cellular zones of the cerebral cortex nor reduced cell density of the cerebral cortex accounts for the alterations in neurochemical parameters observed in Ts19 mice. These results suggest that the effects of the triplication of specific genes on the respective chromosomes, rather than a generalized disruption of developmental homeostasis resulting from extra chromosomal material, may produce selective alterations in neurochemical and neuroanatomical markers observed in these two mouse trisomies.
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    Developmental Genetics 8 (1987), S. 305-320 
    ISSN: 0192-253X
    Keywords: maize ; chlorophyll-deficient mutants ; high-chlorophyll-fluorescent mutants ; albino mutants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Although a wide range of mutations in the nuclear genome also affect chloroplast biogenesis, their pleiotropic nature often limits their use in studying nuclear genes that regulate or facilitate chloroplast development. However, many mutations that cause a high-chlorophyll-fluorescent (hcf) phenotype exhibit limited pleiotrophy, causing the loss of functionally related sets of chloroplast polypeptides. Several hcf mutations are described that result in the loss of one specific protein complex from the thylakoid membrane. Chlorplast and cytosolic mRNAs coding for component polypeptides of the missing complex are unaffected in the mutants, suggesting that each mutation disrupts some process in the synthesis and assembly of the missing complex. Another hcf mutation causes both the loss of three protein complexes and grossly abnormal thylakoid membrane structures. The primary effect of this mutation might be in the assembly of thylakoid membranes or in the stable accumulation of the three protein complexes. Two other hcf mutations are more pleiotropic. Hcf*-38 causes a quantitative reduction of many chloroplast proteins and a reduction of some chloroplast RNAs, including several splicing intermediates. Hcf*-7 causes a major reduction of all chloroplast-encoded proteins examined. The range of pleiotropic effects of hcf mutations indicates that the mutations identify nuclear genes whose products are involved in a number of different steps in chloroplast devclopment. Because some of the mutations described have been generated by transposon insertions, they can be cloned using the transposon to identify the mutant allele.
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    Developmental Genetics 8 (1987), S. 389-403 
    ISSN: 0192-253X
    Keywords: nuclear mutations ; chloroplast assembly ; maize ; light ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The major chlorophyll a/b light harvesting complex (LHCII) of mesophyll chloroplasts is normally assembled late during chloroplast morphogenesis. LHCII occurs at greatly reduced levels in bundle sheath chloroplasts of maize. In order to understand the normal regulatory mechanisms we are examining nuclear maize mutants that alter either (1) the assembly timing or (2) the steady state level of LHCII in mature mesophyll thylakoids. We have found a delayed greening mutant, v24 (on chromosome arm 2L), that unmasks a second unlinked locus, Mof*, that can mediate LHCII assembly timing. The polypeptides of LHCII are encoded by the nuclear multigene cab family. We find that two alleles at Mof* regulate the steady state level of cab mRNA in parallel to their effect on LHCII assembly timing: The genotype Mof*-1 Mof*-1 v24 v24 corresponds to reduced cab mRNA and late LHCII assembly timing, while Mof*-2 Mof*-2 v24 v24 corresponds to reduced cab mRNA and late LHCII assembly timing. A second group of mutations (Oy-700, pg11 and pg12 reduces LHCII levels in mesophyll thylakoids. This is the first report that pg11 and pg12) reduce the LHCII of mesophyll thylakoids. The basis of pg11 and pg12 is unknown. Mutations at the Oy locus block the chlorophyll biosynthetic enzyme, protopor-phyrin IX Mg-chelatase. Heterozygotes of the codominant mutation Oy-700 with the normal allele (Oy) have reduced LHCII. We have defined genetic backgrounds that suppress and those that do not suppress the Oy-700 Oy phenotype under certain conditions: (1) reduced light intensities (200 μE cm-2 sec-1) and/or (2) plant maturity.
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    Developmental Genetics 8 (1987), S. 475-493 
    ISSN: 0192-253X
    Keywords: methylation ; Adh1 ; Zea ; Arabidopsis ; transformed DNA ; CpG-rich islands ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Higher plant DNA is extensively methylated, but the two methylated sequences (CpG and CpNpG) show different characteristics. Using sequence analysis techniques, we demonstrate that while CpG methylation follows the existing models for cytosine methylation in animals, CpNpG methylation does not. Although there is evidence to support the suggestion that the low CpG frequency has arisen from deaminational conversion of 5-methylcytosine to thymidine, there appears to be no comparable conversion of 5-methylcytosine in the CpNpG configuration. It therefore appears that between the evolution of CpG and CpNpG cytosine methylation systems, a mechanism evolved for the correction of C→T conversion, probably using the methylated strand to direct the repair in the correct direction.
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    Developmental Genetics 9 (1988), S. 13-22 
    ISSN: 0192-253X
    Keywords: ontogeny ; lethality ; gene expression ; mRNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cadmium is a toxic metal that induces the expression of metallothionein genes in many tissues and that binds avidly to metallothionein, a soluble transition metal binding protein. The present study examined the temporal pattern and magnitude of accumulation of metallothionein mRNA in liver of C57BL/6J mice of various ages treated with cadmium. In adult female mice, accumulation was dependent on the dosage level of cadmium and related to the concentration of this metal in liver. The accumulation of metallothionein mRNA in liver depended on age at exposure to cadmium. Intraperitoneal administration of 2 mg of cadmium per kg provoked small increases (two- to threefold) in levels of metallothionein mRNA in livers of 7- and 14-day-old mice. In contrast, cadmium treatment of 28- and 56-day-old mice resulted in 12- to 19-fold increases in levels of metallothionein mRNA in liver with maximum increases occurring 3 to 4 hr after treatment. Because similar patterns for the accumulation of cadmium of liver were found in 7-, 28-, and 56-day-old mice, observed age-dependent differences in induction of metallothionein mRNA in liver were probably not due to differences in the accumulation of cadmium in this organ. Taken together, these data suggest that tissue-specific factors controlling the expression of metallothionein genes may account for developmental variation in the inducibility of these genes by cadmium. Ontogenic variation in accumulation of metallothionein mRNA after cadmium treatment may be a factor in developmental variation in the acute lethality of cadmium in C57BL/6J mice.
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  • 66
    ISSN: 0192-253X
    Keywords: Daucus carota ; auxin ; gene expression ; mutant ; filtration-enrichment procedure ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cell cultures of the carrot Daucus carota are a useful experimental system for studying the genetic regulation of plant embryogenesis. A modified filtration-enrichment procedure was used to isolate 21 temperature-sensitive variants in somatic embryogenesis; the variants display normal embryo development at the permissive temperature (24°C) and altered development at the restrictive temperature (33°C). Temperature-shift experiments were performed on these variants to determine the timing of gene action for the putative temperature-sensitive alleles. According to their phenotypes at the restrictive temperature, these variants can be divided into six classes: No Growth, Callus Proliferation, Globularstage Block, Oblong-stage Block, Lateral Growth, and Root Formation. Although many variants exhibit lengthy temperature-sensitive periods, the temperature sensitivity of some variants is restricted to one or two embryonic stages. These results plus those in the literature are incorporated into a preliminary model concerning the genetic regulation of carrot embryogenesis.
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    Developmental Genetics 9 (1988) 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 9 (1988), S. 155-165 
    ISSN: 0192-253X
    Keywords: embryonic lethal ; agouti locus ; trophectoderm ; inner cell mass ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The tissue specificity of the lethal yellow mutant was investigated by separation of blastocyst tissues. Embryos from experimental (Ay/ae × Ay/ae) and control (ae/ae × Ay/ae) crosses of the AG/CamPa inbred strain were recovered at 3.5 days post coitum, cultured for 24 hours, and then mechanically dissected into the component tissues of the blastocyst, the inner cell mass (ICM), and trophectoderm. These fragments were then cultured separately, with or without a feeder layer of inactivated fibroblasts, for an additional 3-5 days. Comparisons between experimental and control crosses indicated that the lethal Ay/Ay embryos were among the blastocysts successfully dissected but that both the ICM and trophectoderm from lethal embryos failed to develop further in vitro, eithal with or without feeders.With retrospective identification of the lethal embryos, it was found that at 4.5 days, after 1 day of culture, they had formed morphologically normal blastocysts but were frequently more fragile upon dissection and had smaller ICMs. Although none had hatched from the zona pellucida, some had ruptured it and were halfway out. With culture, lethal ICMs showed no development, and lethal trophectoderm usually attached but showed very limited outgrowth. Thus, no rescue of lethal tissue was shown with dissection and in vitro culture, and results are consistent with the gene affecting both tissues of the late blastocyst.
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    Developmental Genetics 9 (1988) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 70
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    Developmental Genetics 9 (1988), S. 715-732 
    ISSN: 0192-253X
    Keywords: Regulator of postbithorax ; homeosis ; pattern formation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genetic analysis has shown that the gap segmentation gene hunchback (hb) is a member of the genetic hierarchy involved in pattern formation in Drosophila. To identify the hb gene, we have mapped the position of hb mutant breakpoints within a chromosomal walk of the 85A region by genomic Southern blots and determined the transcription pattern of DNA from the walk. We detect a single gene within the domain defined by breakpoint mapping. We conclude that we have identified the hunchback gene because three mutations that inactivate hb physically interrupt or delete this gene. Northern analysis shows that the hb gene gives rise to at least five overlapping transcripts ranging in length from 2.6 to 3.5 kilobases. S1 nuclease and primer extension experiments demonstrate that the gene employs two promoters and three polyadenylation sites. The two hb promoters have different temporal specificities. Transcripts arising from the upstream promoter are detected from 0-12 hours of embryogenesis as well as in adult female and male RNA preparations. Transcripts arising from the downstream promoter accumulate only from 0-6 hours of embryogenesis. During the syncytial blastoderm stage, transcripts from the hb gene accumulate over a broad anterior and a narrow posterior domain. This pattern sharpens during the late blastoderm/early gastrula stage to produce an embryo with two stripes of hybridization anterior and one stripe posterior. Later, hb transcripts are detected within the ventral hypoderm in extended germ band stage embryos.
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    Developmental Genetics 9 (1988), S. 733-741 
    ISSN: 0192-253X
    Keywords: Phycomyces blakesleeanus ; developmental mutants ; phorogenesis ; sexual reproduction ; light ; carotene ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The mycelium of the fungus Phycomyces. essentially a giant multinucleate cell, produces two kinds of asexual reproductive structures, called macrophores and microphores, and a succession of structures for sexual reproduction. Following the treatment of spores with N-methyl-N′ -nitro-N-nitrosoguanidine, conditional imb mutants have been isolated that form no macrophores at 26°C, but do at 14°C. At the restrictive temperature, few imb mutants (2 of 13) develop microphores, and none is able to complete the sexual cycle. This suggests that genes responsible for macrophorogenesis are involved in microphorogenesis and in sexual development as well. Light reduces macrophorogenesis and totally abolishes microphorogenesis in the wild type under the conditions of our experiments. These photomorphogenetic effects require the normal function of genes madA and madB, which are responsible for phototropism. Light inhibits microphorogenesis in the two imb mutants that form microphores at the restrictive temperature. Genetic alterations of carotenogenesis lead to an excess of microphores and a scarcity of macrophores in the dark, but they have little influence on vegetative reproduction in the light.
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    Developmental Genetics 9 (1988), S. 215-226 
    ISSN: 0192-253X
    Keywords: cAMP ; cGMP ; chemotaxis ; mutant ; desensitization ; receptor ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The functional interaction of surface cAMP receptors with effector enzymes via G-proteins was investigated in Dictyostelium discoideum. Several experimental conditions were used to investigate signal transduction, such as reduced temperatures, use of down-regulated cells and of mutants. The results are presented as a model describing the complex interaction between multiple forms of the surface cAMP receptor and different G-proteins that are responsible for the generation of the second messengers, cAMP, cGMP, InsP3 and Ca2+.
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    Developmental Genetics 9 (1988), S. 227-235 
    ISSN: 0192-253X
    Keywords: receptors ; transmembrane signalling ; Dictyostelium ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using antibodies specific for the 3′, 5′-cyclic AMP (cAMP) cell surface receptor of Dictyostelium discoideum, we have screened γgtll expression libraries and isolated a series of cDNAs derived from cAMP receptor mRNA during early development. The identity of the cDNA clones was verified by multiple criteria: (1) β-galactosidase fusion proteins synthesized by isolated cDNA clones stain intensely with cAMP receptor directed antiserum, (2) these fusion proteins affinity purify antibodies specific for the cAMP receptor, (3) the cDNA probes hybridize to a 2 kb mRNA whose change in relative level of abundance during development parallels that of receptor mRNA as assayed by in vitro translation, (4) the 2 kb mRNA size equals that of receptor mRNA as determined by in vitro translation of size fractionated poly (A)+ RNA, and (5) RNA transcribed in vitro from cDNAs containing the entire protein-coding region produces a polypeptide by in vitro translation with an apparent molecular weight in close agreement with that of nascent cAMP receptor protein produced by in vitro translation of cellular RNA.The DNA sequence predicts an open reading frame of 392 amino acids. The deduced amino acid sequence contains seven domains enriched in hydrophobic residues. A model is proposed in which the cAMP cell-surface receptor traverses the lipid bilayer seven times in a pattern similar to that of other receptors, such as rhodopsin, which interact with G-proteins. The structural similarities suggest a gene family of related surface receptors from such evolutionarily diverse species as Dictyostelium, yeast, and mammals.
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    Developmental Genetics 9 (1988), S. 247-258 
    ISSN: 0192-253X
    Keywords: protein evolution ; lower eukaryote ; differentiation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cAMP-dependent protein kinase (cAK) from Dictyostelium discoideum is an enzyme composed of one catalytic and one regulatory subunit. Upon binding of cAMP, the holoenzyme dissociates to liberate free active catalytic subunits. The cAK is developmentally regulated, ranging from very little activity in vegetative cells to maximal expression in postaggregative cells. Although there is no immunological cross-reaction between the subunits of cAKs from Dictyostelium and from other organisms, they share several biochemical properties. A complete cDNA for the regulatory subunit has been cloned and sequenced. Only one copy of the gene for the regulatory subunit is present per haploid genome. On the basis of the comparison of the structure of the cAK from Dictyostelium with its counterparts in yeast and higher eukaryotes, we propose a model for the evolution of cyclic-nucleotide-binding proteins.
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  • 75
    ISSN: 0192-253X
    Keywords: gene regulation ; immunoblotting ; rapid-developing variants ; molecular cloning ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Several lines of evidence indicate that cAMP modulates developmental gene activity via cell-surface receptors. We describe here a novel cAMP receptor, CABP1, whose properties are consistent with the idea that this protein is involved in gene regulation. Firstly, immunological techniques using anti-CABP1 antibodies as probes showed that this cAMP receptor can be detected on the surface of developing cells. Secondly, there is a steady migration of CABP1 to the nucleus during development. Thirdly, some genetic variants exhibiting an altered pattern of development are found to possess modified CABP1. We also showed that CABP1 co-purifies with at least seven other polypeptides which share common epitopes with CABP1. Interestingly, four of the CABP1-related polypeptides can be detected on the cell surface as well as in the nucleus.
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  • 76
    ISSN: 0192-253X
    Keywords: transformation ; gene structure ; cAMP ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cyclic nucleotide phosphodiesterase (phosphodiesterase) of Dictyostelium discoideum plays an essential role in development by hydrolyzing the cAMP used as a chemoattractant by aggregating cells. We have studied the biochemistry of the phosphodiesterase and a functionally related protein, the phosphodiesterase inhibitor protein, and have cloned the cognate genes. A 1.8-kb and a 2.2-kb mRNA are transcribed from the singlephosphodiesterase gene. The 2.2-kb mRNA comprises the majority of the phosphodiesterase mRNA found in differentiating cells and is transcribed only during development from a promoter at least 2.5 kb upstream of the translational start site. The 1.8-kb phosphodiesterase mRNA is detected at all stages of growth and development, is present at lower levels than the developmentally induced mRNA, and is transcribed from a site proximal to the protein-coding region. The phosphodiesterase gene contains a minimum of three exons, and a 2.3-kb intron, the longest yet reported for this organism. We have shown that the pds A. gene and fourfgd genes affect, the accumulation of the phosphodiesterase mRNAs, and we believe that these loci represent a significant portion of the genes regulating expression of the phosphodiesterase. The phosphodiesterase gene was introduced into cells by transformation and used as a tool to explore the effects of cAMP on the terminal stages of development. In cells expressing high levels of phosphodiesterase activity, final morphogenesis cannot be completed, and differentiated spore and stalk cells do not form. We interpret these results to support the hypothesis that cAMP plays an essential role in organizing cell movements in late development as well as in controlling the aggregation of cells in the initial phase of the developmental program.
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    Developmental Genetics 9 (1988), S. 259-265 
    ISSN: 0192-253X
    Keywords: G-proteins ; gene expression ; developmental regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have identified a two-member gene family in the Dictyostelium genome and have isolated corresponding cDNA or genomic DNA recombinant clones. Analyses of these DNA sequences predicted encoded proteins of ∼200 amino acids with ∼90% sequence identity to each other. These Dictyostelium proteins also share amino acids identity within the GTP-binding domains in the family of G-regulatory proteins involved in cellular regulation and transmembrane signalling. Additional structural similarities are seen with members of the ras supergene family, such as ras, ral, and rho. They are similar in size (usually ∼200 amino acids), possess four conserved domains involved in GTP interaction and are believed to be anchored in the membrane by fatty acid modification of a cysteine residue near the carboxy terminus. More extensive identity is observed with YPT1 and SEC4, two other members of this family of genes that are essential in yeast. The amino-terminal half of both Dictyostelium proteins is 70% identical in amino acid sequence to the YPT1 and SEC4 yeast proteins with less identity continuing through the remainder of the proteins. In addition these proteins terminate in two cysteine residues that are thought to be required for membrane anchorage.The two genes within this Dictyostelium family are organized differently in the genome and are differentially regulated during development. One gene is colinear in sequence with its mRNA in the protein coding region, whereas the other gene encodes a spliced mRNA. The intron-containing gene is associated with a developmentally regulated (AAC)-repeat sequence. Finally, we have shown that the expression of one of the genes is induced during development with kinetics similar to that of other (AAC)n-associated genes; conversely, the expression of the second gene is repressed at a similar developmental stage.
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  • 78
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    Developmental Genetics 9 (1988), S. 303-313 
    ISSN: 0192-253X
    Keywords: developmentally regulated cDNAs ; Dictyostelium discoideum gene sequences ; developmentally regulated proteins ; Dictyostelium spore proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Similar to other stages in Dictyostelium development, spore germination is a particularly suitable model for studying the regulation of gene expression, because developmentally regulated changes in both protein and mRNA synthesis occur during the transition from dormant spore to amoeba. Spores are constitutively dormant and must be activated to germinate. Under the proper environmental conditions, spores germinate in a highly synchronous manner to give rise to individual amoebae that can then enter the vegetative growth phase. Protein synthesis is developmentally regulated during this process. Because protein synthesis is transcriptionally controlled during spore germination, the respective genes must be developmentally transcribed, and these can be isolated and analyzed. Three cDNA clones specific for mRNA developmentally regulated during spore germination have been characterized and used as probes to study mRNA accumulation and decay during spore germination. Because we are interested in defining the sequences of developmentally regulated genes that may relate to their regulation of transcription, we have sequenced the cDNAs and have isolated and sequenced their respective genomic clones. The sequences of the three gene families, their genomic organization, and their special structural features are described.
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  • 79
    ISSN: 0192-253X
    Keywords: ions ; developmental regulation ; receptor regulation ; developmental signals ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have analyzed the effects of the cAMP relay inhibitor, caffeine, and the receptor antagonist, adenosine, on the regulation of the cell-surface cAMP receptor in suspensionstarved Dictyostelium discoideum cells by measuring ammonium sulfate-stabilized binding of [3-H]cAMP to intact cells. When cells were starved in fast (230 r.p.m.) shaken suspension in 10 mM Na+/5 mM K+ phosphate buffer, pH 6.5, plus 1 mM CaCl2 and 2.5 mM MgCl2, and assayed for specific cAMP binding, receptor accumulation peaked at approximately 6 hours, reaching a maximum of 1.5 pmol cAMP bound/107 cells (saturation binding). Neither caffeine nor adenosine inhibited the accumulation of cAMP receptors. Similar results were obtained in caffeine-treated, slow shaken (90 r.p.m.) suspension cultures. These results suggest that starvation alone is sufficient stimulus to induce the cAMP receptor. We have also tested the effects of different buffer ionic compositions on the accumulation of cAMP receptors. Elevation of the monovalent ion concentration to 30-40 mM was found to significantly inhibit the induction of cAMP receptors.
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  • 80
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    Developmental Genetics 9 (1988), S. 279-292 
    ISSN: 0192-253X
    Keywords: Dictyostelium ; development ; cAMP ; cell-cell contact ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: cAMP and cell-cell contact are involved in the coordination of differentiation and morphogenesis in Dictyostelium discoideum. The experiments described in this paper establish a relationship between cAMP and cell-cell contact. Contact between Enterobacter aerogenes and aggregation-competent Dictyostelium amoebae and contact between Dictyostelium amoebae themselves results in the transient secretion of cAMP and an alteration in the amount of cAMP secreted in response to subsequent stimulation by cAMP, i.e., an alteration in magnitude of a cAMP relay response. The subsequent cAMP relay response can be enhanced or diminished depending upon the number of contacts formed and the concentration of cAMP present at the time of contact. Latex beads are capable of evoking cAMP secretion. However, the bead/amoebal contact is unable to alter the magnitude of a subsequent response to cAMP. This suggests that a nonspecific interaction via cell-cell contactelicits transient cAMP secretion in aggregation-competent Dictyostelium amoebae.The two responses to cell-cell contact are distinct from each other and distinct from the cAMP relay response. (1) The dose-response curves for the responses to Enterobacter contact are clearly different. (2) Contact with latex beads can elicit cAMP secretion but not alter the magnitude of a subsequent cAMP relay response. (3) The temperature dependences of the contact-induced responses and the cAMP relay response show that only the contact-induced cAMP secretion is inhibited at 12 and 15°C, while only the cAMP relay response is inhibited at 28°C.A 4-second application of cAMP at the time that contact is initiated enhances both contact-induced responses. Whether the relationship between these two developmental regulators is important for the regulation of Dictyostelium development has yet to be established.
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  • 81
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    Developmental Genetics 9 (1988), S. 327-335 
    ISSN: 0192-253X
    Keywords: gene regulation ; initiation of development ; slime mold ; transcription ; cycloheximide ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Several genes that are deactivated upon the initiation of development of Dictyostelium discoideum have been identified by differential screening of various cDNA libraries. These genes have in common a decrease in the steady-state levels of their corresponding mRNAs as development proceeds. When development was carried out in the absence of protein synthesis by inhibition with cycloheximide, the decrease in mRNA levels for most genes (V genes) was normal or slightly accelerated. However, for about 5% of the genes (H genes), cycloheximide caused an apparent induction of expression, as revealed by a slight or dramatic increase in mRNA levels instead of the normal decrease. This effect was due to inhibition of protein synthesis and not to cycloheximide per se. The induction was found to be due to an enhancement of the trascription rate; normal rates of transcription for the H genes were dependent upon continued protein synthesis during vegetative growth and during development. Thus, two general regulatory classes exist for deactivation of gene expression upon initiation of development, one dependent and one independent of protein synthesis. Models concerning the control of expression of these two classes of genes are discussed here. Analysis of expression of these genes in mutant strains that are aggregation-deficient has also been performed, and the results lead to subdivisions of the classes.
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  • 82
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    Developmental Genetics 9 (1988), S. 315-326 
    ISSN: 0192-253X
    Keywords: Dictyostelium ; growth ; development ; regulation ; discoidin I ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have previously shown that growing cells of Dictyostelium discoideum (strains NC4 and AX3) produce a soluble substance that accumulates in the medium in proportion to cell density; this substance regulates the production of certain proteins previously thought to be induced by starvation [Clarke et al., 1987]. We suggest the name PSF (prestarvation factor) for this substance. During growth, Dictyostelium cells monitor the relative concentrations of PSF and food bacteria. When PSF reaches a sufficiently high level relative to the concentration of bacteria, synthesis of PSF-regulated proteins is induced. We propose the name prestarvation response for this induction, which takes place in exponentially growing cells several generations before the food bacteria are depleted. We have explored the mechanism by which the food bacteria inhibit the response of Dictyostelium cells to PSF. We find that the bacteria do not inactivate PSF or inhibit its production; instead, they affect the ability of NC4 cells to detect PSF, possibly by binding to the same cell surface receptor. In the absence of bacteria, as during axenic growth of AX3 cells, the prestarvation response occurs at much lower cell densities, probably accounting for the presence of certain developmentally regulated mRNAs and proteins in axenic cultures.
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  • 83
    ISSN: 0192-253X
    Keywords: cAMP ; Ca2+ ; signal transduction ; cell surface receptor ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Elevated levels of cAMP are essential for the expression of many postaggregation prespore and prestalk mRNA species and for the suppression of some growth phase mRNAs. Here we review evidence that this regulation is mediated by cAMP interacting at the cell surface receptor. These effects of cAMP on gene expression can occur under conditions where the receptor-associated adenylate cyclase is inactivated and in concentrations that are consistent with receptor binding. A number of differences are noted in the mechanism by which cAMP regulates prespore and prestalk genes. Finally, evidence is reviewed for the role of a Ca2+-dependent signal transduction system in coupling the expression of some of the prespore mRNAs to the cAMP receptor. This signal transduction system does not appear to be involved in the expression of the cAMP-dependent prestalk gene.
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  • 84
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    Developmental Genetics 9 (1988), S. 337-350 
    ISSN: 0192-253X
    Keywords: Dictyostelium ; cAMP ; receptor ; gene regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have examined the expression of a cAMP pulse-repressed and two cAMP pulse-induced genes in response to cAMP and caffeine under a number of different physiological conditions, and in several classes of developmental mutants altered in cAMP-mediated signal transduction pathways. The data presented help characterize the mutants with regard to early gene expression. Analysis of the data indicates that full induction of the pulse-induced or repression of the pulse-repressed genes requires cycles of activation and adaptation of the cAMP receptor but does not require a rise in intracellular cAMP. Comparison of the results obtained between different mutant classes suggests that repression and activation of the two classes of genes can be uncoupled, implying that different intracellular mechanisms control these processes. In addition, we examined the effects of caffeine and show that it can induce pulse-induced mRNA accumulation in the absence of cAMP.
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  • 85
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    Developmental Genetics 9 (1988), S. 351-358 
    ISSN: 0192-253X
    Keywords: Dictyostelium ; diacylglycerol ; inositol trisphosphate ; developmetal regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Dictyostelium, extracellular cAMP interacts specifically with cell-surface receptors to promote the accumulation of a variety of intracellular second messengers, such as, 3′-5′ cyclic adenosine monophosphate (cAMP) and 1,4,5 inositol trisphosphate (IP3). We and others have shown that activation of the cell-surface cAMP receptor can also modulate the expression of the Dictyostelium genome during development. In at least one instance, synthesis of intracellular cAMP is required for appropriate gene regulation. However, the induction of most cAMP-dependent gene expression can occur in the absence of receptor-mediated activation of adenylate cyclase and a consequent accumulation of intracellular cAMP. These results suggest that other intracellular second messengers produced in response to receptor activation may potentially act as signal transducers to modulate gene expression during development. In vertebrate cells, IP3 and diacylglycerol (DAG) are intracellular activators of specific protein kinases; they are produced in equimolar amounts by cleavage of phosphoinositol bisphosphate after a receptor-mediated activation of a membrane-bound phosphodiesterase. IP3 and, thus, by inference, diacylglycerol are synthesized in Dictyostelium as a response to cAMP interacting with its cell-surface receptor. Using defined conditions to inhibit the accumulation of extracellular cAMP, we have examined the effects of these compounds on the accumulation of extracellular cAMP, we have examined the effects of these compounds on the expression of genes that require cAMP for their maximal expression. Our results suggest that intracellular IP3 and DAG may in part mediate the action of extracellular cAMP on the expression of the Dictyostelium genome.
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  • 86
    ISSN: 0192-253X
    Keywords: Dictyostelium cell type markers ; PsA ; phosphatidylinosito ; D19 gene ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of D19, a Dictyostelium gene that encodes a prespore-specific mRNA sequence shows it to encode PsA, the cell surface protein detected by the MUD 1 monoclonal antibody. The predicted sequence of the protein reveals a largely hydrophobic C terminus, with chemical similarity to proteins known to be attached to the plasma membrane via a phosphatidylinositol link. The C-terminal region has direct sequence homology to the contact sites A protein and to the phosphatidylinositol-linked form of a chicken N-CAM, suggesting that it might play a role in cell adhesion. Expression of the D19 gene is known to be induced by cAMP and repressed by adenosine. The accumulation of the D19 mRNA is also repressed by DIF, the putative stalk-specific morphogen, and this effect is mediated at the transcriptional level. The pDd56 and pDd63 genes are induced by DIF, and they are specific markers of prestalk and stalk cells. They encode, respectively, ST310 and ST430, two proteins that were first identified by two-dimensional gel electrophoresis. Both proteins are predominantly composed of a highly conserved, 24-amino acid repeat. The two proteins are localized in the slime sheath of the migratory slug and in the stalk tube and stalk cell wall of the mature culminant, where they presumably function as structural components of the extracellular matrix. We have constructed marked derivatives of the pDd56, pDd63, and D19 genes, and these are correctly regulated after transformation into Dictyostelium cells. Thus we have determined the structure, and elucidated possible functions, for one prespore and two prestalk genes. These sequences should be of value, both as markers of the earliest events in cellular differentiation and in identifying the regulatory sequences controlling cell type-specific gene expression.
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  • 87
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    Developmental Genetics 9 (1988), S. 371-382 
    ISSN: 0192-253X
    Keywords: second messenger ; transcription ; DNase I hypersensitive sites ; cis-acting elements ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: cAMP regulates gene expression in Dictyostelium discoideum through the cell surface receptor and is therefore a transmembrane signal transduction event. We have now begun to examine the signal transduction pathway that transmits the cAMP-induced signal to the nucleus. The results presented here indicate that Ca2+ plays a crucial role. A comparison of the accumulation of UDPGP1 mRNA during development with the corresponding transcription rates revealed that this gene is regulated primarily at the level of transcription. To elucidate the factors involved in the regulation of the UDPGP1 gene we characterized its cis acting sequences. We constructed a series of deletions into the 5′ flanking region of the UDPGP1 gene and analyzed the expression of the mutated DNA in transformants. A sequence element essential for the expression of the UDPGP1 gene is located between -500 bp and -288 dp from the transcription start site. This promoter element appears to be a short G + C-rich sequence positioned between -374 to -395 and coincides with a DNase I hypersensitive site.
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  • 88
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    Developmental Genetics 9 (1988), S. 403-419 
    ISSN: 0192-253X
    Keywords: post-transcriptional regulation ; disaggregation ; poly(A)-binding proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This paper reviews our studies of three aspects of post-transcriptional regulation in Dictyostelium discoideum: (1) the determinants of mRNA stability in vegetative amoebae; (2) the effects of disaggregation and cyclic AMP on the decay rates of cell-type-specific mRNAs in late developing cells; and (3) the cytoplasmic function of the 3′ poly(A) tracts present on most mRNAs. We find that: (1) mRNA stability in vegetative amoebae is not dependent on mRNA size, ribosome loading, or poly(A) tract length, but may be determined by specific 3′-untranslated sequences within a given mRNA; (2) mRNA decay rates in late developing cells are heterogeneous, and cyclic AMP does not act directly to stabilize cell-type-specific mRNAs; and (3) poly(A) is most likely involved in the initiation of protein synthesis via an interaction with cytoplasmic poly(A)-binding proteins.
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  • 89
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    Developmental Genetics 9 (1988), S. 421-434 
    ISSN: 0192-253X
    Keywords: translational control ; polyadenylation ; mRNA stability ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated recombinant plasmids that contain cDNA inserts complementary to mRNAs encoding six different r-proteins of Dictyostelium discoideum. Southern and quantitative dot blot analyses have shown that each of the r-protein genes represented in these plasmids is encoded by a single copy gene and that these genes are not tightly linked to each other. We have determined the relative amount of the six r-protein mRNAs present in cells at intervals throughout development and find that for the first 9 hours of development, each of the mRNAs remains present at virtually the same level as in vegetatively growing cells. Between 9 and 11 hours of development, there is a rapid loss of these mRNAs to 15% or less of vegetative levels, and that low level remains, or slightly declines, through the late stages of development. We have shown that two post-transcriptional events contribute to the developmental regulation of the expression of the r-protein genes. The first involves a specific block to translational initiation that is not the result of inactivation of these mRNAs by decapping or deadenylation. The second is a change in the stability of these mRNAs during early development. In order to begin to analyze the role of specific sequences that may act as targets or signals in these events, we have cloned and sequenced a 1.9-kb genomic DNA fragment that encodes one of the r-proteins. We find that transcription of this gene begins in a pyrimidine-rich region that is not preceded by a TATA box, the gene contains a single intron of 350 bp, and there are two alternative 3′ processing sites. In addition, the 5′-untranslated region of the transcript contains an unusually high percentage of G and C residues relative to other Dictyostelium mRNAs.
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  • 90
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    Developmental Genetics 9 (1988), S. 597-605 
    ISSN: 0192-253X
    Keywords: cell differentiation ; differentiation inducing factor ; cyclic AMP ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Stalk cell formation in low-cell-density monolayers of Dictyostelium discoideum, strain V12-M2, occurs following the sequential addition of cyclic AMP and the differentiation-inducing factor (DIF). Both cyclic AMP and DIF are essential for the appearance of the prestalk-specific isozyme alkaline phosphatase-II, which suggests that both factors are necessary for prestalk cell formation. The available evidence suggests that the cyclic AMP requirement for stalk cell formation is mediated through the cell surface cyclic AMP receptor. However, stalk cell formation is inhibited by caffeine and this inhibition is reversed by the cell-permeable analogue 8-Br-cyclic AMP, which suggests in addition a possible involvement for elevated intracellular cyclic AMP concentrations in stalk cell formation.During in vivo development ceils first become independent of cyclic AMP at the tipped aggregate stage, but the acquisition of cyclic AMP independence is advanced by several hours when cells are incubated in the presence of cyclic AMP for 2 hours. Cells do not become independent of DIF until the culmination stage of development, which suggests the possibility that DIF is required for the conversion of prestalk cells to stalk cells.There is an absolute requirement for DIF for stalk cell formation in low-density monolayers of prestalk cells but only part of population exhibits a requirement for cyclic AMP, which suggests that the prestalk cell population consists of two distinct cell types. Stalk cell formation from prespore cells is totally dependent on both cyclic AMP and DIF.When isolated prestalk and prespore cells are plated in high-density monolayers, the former cells accumulate more DIF. which suggests the possibility that DIF is preferentially synthesized by prestalk cells.
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  • 91
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    Developmental Genetics 9 (1988), S. 615-628 
    ISSN: 0192-253X
    Keywords: colony morphology ; yeast cell wall ; aggregation variants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recently, high frequency switching systems have been identified in the infectious yeast Candida albicans and the cellular slime mold Dictyostelium discoideum. In C. albicans, cells can switch at spontaneous frequencies as high as 10-2 between seven general colony morphologies in the case of strain 3153A or between two major phenotypes in the white-opaque transition in strain WO-1. In the latter system, dramatic changes occur in cellular phenotype as well. In D. discoideum, cells can switch at spontaneous frequencies of roughly 10-2 between a number of colony phenotypes which include alterations in developmental timing, blocks at particular morphogenetic stages, morphological aberrations, and aggregation-minus. In the C. albicans and D. discoideum switching systems, the following characteristics are shared: (l) a limited number of switch phenotypes; (2) heritability; (3) high frequency reversibility; (4) low and high frequency modes of switching; and (5) ultraviolet (UV) stimulation of switching of cells in a low frequency mode of switching.
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  • 92
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    Developmental Genetics 9 (1988), S. 607-614 
    ISSN: 0192-253X
    Keywords: pattern formation ; cell sorting ; differential chemotaxis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: When cells dissociated from Dictyostelium discoideum slugs were cultured in roller tubes, they formed agglomerates in which prestalk cells were initially dispersed but soon sorted out to the center and then moved to the edge to reconstitute the prestalk/prespore pattern. To examine the mechanism of sorting out, individual prestalk cells were traced by a videotape recorder. The radial component of the rate of movement toward the center of the presumptive prestalk region was calculated. Prestalk cells did not move randomly, but rather directionally toward the center. Thei movement was pulsatile, with a period of ca. 15 min, and accompanied by occasional formation of cell streams, thus resembling the movement observable during cell aggregation. These results favor the idea that prestalk cells sort out to the prestalk region due to differential chemotaxis rather than differential adhesiveness. After formation of the prestalk/prespore pattern, the prestalk region rotated along the circumference of the agglomerates. This appears comparable to migration of slugs on the substratum, the rate of rotation being similar to that of slug migration.To examine the processes of pattern formation during development. washed vegetative cells were cultured in roller tubes. Prespore cells identified by antispore immunoglobulin initially appeared randomly within the agglomerates, but then nonprespore cells accumulated in the center and finally moved to the edge to establish the prestalk/prespore pattern, the processes being similar to those of pattern reconstruction with differentiated prestalk and prespore cells.
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  • 93
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    Developmental Genetics 9 (1988), S. 629-638 
    ISSN: 0192-253X
    Keywords: transposon ; cellular slime mold ; development ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: PN6024 is a mutant strain of P. pallidum which appeared on selection for resistance to MDMP, an inhibitor of translation. It was found to be mutant in four other traits, being resistant to tubercidin, incapable of growth at 31.5°C, abnormal in development, and slow growing at 25°C. Genetic crosses using the macrocyst cycle showed that these five traits are controlled by five unlinked genes. The hypothesis is that movement of a transposon to multiple new locations caused these mutations. A difference in restriction fragment pattern between PN6024 and its parent PN600 support the hypothesis. Attempts were made to find conditions generating other strains like PN6024. Selection for growth in the presence of tubercidin yielded clones which resemble PN6024 in being developmentally abnormal as well as tubercidin resistant. Tubercidin treatment also increased the frequency of clones resistant to canavanine. It is suggested that tubercidin is mutagenic because it causes movement of the putative transposon, not because it generates point mutations. Growth under conditions of stress (at 31.5°C, at 8°C, in the presence of 2% ethanol) had at most an erratic effect in generating strains like PN6024.Three substrains appeared spontaneously in cultures of PN6024. These differed in developmental characteristics from each other and from the parent strain. It is suggested that they carry mutations in genes which control the choices between growth and aggregation, and between aggregation and encystment.
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    Developmental Genetics 9 (1988), S. 663-672 
    ISSN: 0192-253X
    Keywords: patterning ; reaction-diffusion ; slime mold ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The formation of secondary sori in whorls of Polysphondylium pallidum provides an attractive model system for the study of symmetry breaking during morphogenesis. Tip-specific antibodies that permit detection of very early stages in this patterning process are available. We have found that the patterns of tip-specific antigen expression vary considerably depending on the size, shape, and developmental stage of the whorl. All of these patterns, however, are well explained by patterning models that rely on short-range autocatalysis and long-range inhibition, as exemplified by reaction-diffusion theories. In the context of reaction-diffusion, we discuss the possible effects of initial conditions, boundary conditions, and nonlinearities on the selection of patterns in P. pallidum whorls.
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  • 95
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    Developmental Genetics 9 (1988), S. 673-681 
    ISSN: 0192-253X
    Keywords: cytokinesis ; phagocytosis ; adhesion ; cytoskeleton ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cellular slime mold amoebae have become a model system for the study of cell motility and the cytoskeleton. A basic problem which all cells face that involves the cytoskeleton is how to control their size. The varied ways in which cellular slime mold amoebae change their cell size-by changing the size at which division occurs, by cell fusion, and by control over cytokinesis-are reviewed. A model is presented which attempts to explain how the mechanisms affected in certain cytokinesis mutants in Dictyostelium discideum known as phg mutants could be involved in control of cell size in the predatory slime mold Dictyostelium caveatum.
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  • 96
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    Developmental Genetics 10 (1989) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 97
    ISSN: 0192-253X
    Keywords: Oogenesis ; Eggshell ; Gene family ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This study isolated cDNA clones from egg-chamber and adult female Drosophila cDNA libraries using as probe a DNA fragment from a 200-kb “chromosome walk” in region 32E of the second chromosome of D. melanogaster. The present authors believe that these clones correspond to a new vitelline membrane protein (VMP) gene because (1) cDNA clones in Northern blots identify a transcript expressed in a tissue- and stage-specific manner: stage 10 egg-chambers; (2) the sequence of cDNAs and of the genomic subclone shows homology with the other VMP genes that have been identified to date; (3) the amino acid composition of the translational product has the high content of proline and alanine characteristic of VMPs. Two aspects emerging from this study are worth stressing: (1) the presence of a hydrophobic domain that is highly conserved in all the VMP genes; and (2) the particularly narrow period of expression of the isolated gene, which could be related to the mechanism of vitelline membrane assembly.
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    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 99
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989), S. 63-69 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989), S. 70-86 
    ISSN: 0192-253X
    Keywords: Arrested cleavage ; Centrosome ; contractile ring ; Fusome ; Germarium ; Models of dividing cells ; Oocyte/nurse cell syncytium ; Ovarian tumor mutation ; POlytrohic meroistic ovary ; Ring canal ; Spindle elongation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three-dimensional models were constructed utilizing the information gained from electron micrographs of serial sections of two clones of cystocytes undergoing their terminal divisions. In each clone a polyfusome connected all eight cystocytes together. Each of the spindles was oriented so that one pole touched the polyfusomes, while the other pointed away from it. This positioning of spindles ensures that one cell of each dividing pair retains all previously formed canals, while the other receives none. The two cells that eventually come to contain the maximum number of canals and fusomal material are the ones that differentiate as pro-oocytes, while the others become nurse cells. The orientation of each spindle suggests that the polyfusome formed at one division determines the placement of the cytoskeletal fibers that anchor the spindles formed at the next division. There is a centripetal gathering together of new canals following each cycle of cystocyte division, which is thought to result from the subsequent contraction of the polyfusomal system. Females homozygous for the otu1 mutation are characterized by ovarian tumors, which result when germarial cystocytes undergo supernumerary divisions and fail to differentiate into either nurse cells or oocytes. An analysis of electron micrographs taken of serially sectioned, mutant germaria showed that most germ cells were single or belonged to clusters of two or three interconnected cells. Therefore otu1 cystocytes are unable to undergo a sustained series of arrested cleavages. These cystocytes contain fusomal material that shows ultrastructural differences from normal polyfusomes. We conclude: (1) that a normal polyfusomal system is a necessary prerequisite for the production of a branched chain of cystocytes and for their subsequent differentiation into pro-oocytes and nurse cells; and (2) that a product encoded by the otu+ gene is essential for the construction of a functional polyfusome.
    Additional Material: 16 Ill.
    Type of Medium: Electronic Resource
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