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  • 101
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 16-28 
    ISSN: 0192-253X
    Keywords: Mesoderm ; fate-mapping ; germ layer formation ; morphogenetic movement ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The developmental fate of cells in the epiblast of early-primitive-streak-stage mouse embryos was assessed by studying the pattern of tissue colonisation displayed by lac Z-expressing cells grafted orthotopically to nontransgenic embryos. Results of these fate-mapping experiments revealed that the lateral and posterior epiblast contain cells that will give rise predominantly to mesodermal derivatives. The various mesodermal populations are distributed in overlapping domains in the lateral and posterior epiblast, with the embryonic mesoderm such as heart, lateral, and paraxial mesoderm occupying a more distal position than the extraembryonic mesoderm. Heterotopic grafting of presumptive mesodermal cells results in the grafted cells adopting the fate appropriate to the new site, reflecting a plasticity of cell fate determination before ingression. The first wave of epiblast cells that ingress through the primitive streak are those giving rise to extraembryonic mesoderm. Cells that will form the mesoderm of the yolk sac and the amnion make up a major part of the mesodermal layer of the midprimitive-streak-stage embryo. Cells that are destined for embryonic mesoderm are still found within the epiblast, but some have been recruited to the distal portion of the mesoderm. By the late-primitive-streak-stage, the mesodermal layer contains only the precursors of embryonic mesoderm. This suggests that there has been a progressive displacement of the midstreak mesoderm to extraembryonic sites, which is reminiscent of that occurring in the overlying endodermal tissue. The regionalisation of cell fate in the late-primitive-streak mesoderm bears the same spatial relationship as their ancestors in the epiblast prior to cell ingression. This implies that both the position of the cells in the proximal-distal axis and their proximity to the primitive streak are major determinants for the patterning of the embryonic mesoderm. © 1995 Wiley-Liss, Inc.
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  • 102
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 65-77 
    ISSN: 0192-253X
    Keywords: Follistatin ; activin ; inhibin ; chick ; rhombomeres ; somites ; resegmentation ; neural induction ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Follistatin, a secreted glycoprotein, has been shown to act as a potent neural inducer during early amphibian development. The function of this protein during embryogenesis in higher vertebrates is unclear, and to further our understanding of its role we have cloned, sequenced, and performed an in-depth expressional analysis of the chick homologue of follistatin. In addition we also describe the expression pattern of activin βA and activin β B, proteins that have previously been shown to be able to interact with follistatin. In this study we show that the expression of follistatin and the activins do not always overlap. Follistatin was first detected in Hensen's node and subsequently in the region described by Spratt [1952] as the neuralising area. In older embryos it was also expressed in a highly dynamic manner in the hind-brain as well as in the somites. We also present evidence that follistatin may have a later role in the resegmentation of the somites. We were unable to detect the expression of activin βA during early embryogenesis, whereas activin βB was first expressed in the extending primitive streak and subsequently in the neural folds. The results from this study are consistent with a role for follistatin in neural induction but suggest it has additional functions unrelated to its inhibitory actions on activins. © 1995 Wiley-Liss, Inc.
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  • 103
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 107-116 
    ISSN: 0192-253X
    Keywords: Axis formation ; fork head ; gastrulation ; neurulation ; Xenopus ; zebrafish ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Intense efforts have been devoted to the identification of genes that are causatively involved in pattern-forming events of invertebrates and vertebrates. Several gene families involved in this process have been identified. Here we focus on the Xenopus fork head domain gene family. One of its members, XFKHl/Pintallavis/XFD1, has been shown previously to be involved in axial formation, and the expression patterns of the other family members discussed below suggest that they too play a major role in the initial steps of patterning and axial organization. In this report, we describe four Xenopus fork head genes XFKH3, 4, 5, and 6) and analyze the distribution of their transcripts during early development. XFKH3 is expressed in developing somites but not notochord, XFKH4 in forebrain, anterior retina, and neural crest cells, and XFKH5 in a subset of epidermal cells and the neural floor plate. Finally, transcripts of XFKH6 are seen in neural crest-derived cranial ganglia. In addition, we show that at least some of the zebrafish fork head genes might serve a comparable function. Zebrafish zf-FKHl has a similar expression pattern as Xenopus XFKHl/Pintallavis/XFDl. It is transcribed in the notochord and neural floor plate. The polster or “pillow” also shows very high levels of zf-FKHl mRNA. © 1995 Wiley-Liss, Inc.
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  • 104
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 167-175 
    ISSN: 0192-253X
    Keywords: Mammary gland ; fatty acid binding protein ; mammary derived growth inhibitor ; proliferation ; differentiation ; transgenic mice ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cardiac fatty acid binding protein (cFABP) is abundantly expressed in the nondividing, functionally differentiated mammary ephithelium. It is very closely related, if not identical to, a previously described protein termed mammary derived growth inhibitor (MDGI). In vitro studies suggest that low concentrations of diffusible cFABP/MDGI may play a hormone-like role in limiting proliferative activity and promoting functional differentiation of this tissue, but no in vivo data to support this idea have been published. To test this hypothesis, we compared the levels of cFABP mRNA with both the epithelial DNA labelling index and levels of β-casein mRNA in wild-type mice. We also investigated the effect of a precocious experimental increase of cFABP levels in the mammary gland of transgenic mice on the labelling index and β-casein mRNA levels. This was accomplished by expressing a bovine cFABP cDNA under the control of the ovine β-lactoglobulin (BLG) gene promoter. We found that although both the DNA labelling index, β-casein mRNA levels, and cFABP mRNA levels in wild-type mice are developmentally regulated, they do not correlate with each other during early pregnancy in individual mice. Moreover, a three- to fourfold increase of total cFABP mRNA in two transgenic lines did not affect the DNA labelling index or the levels of β-casein mRNA, an established marker of differentiation of the mammary epithelium, at this developmental stage. These data suggest that epithelial DNA synthesis, β-casein gene expression, and expression of the cFABP gene are regulated independently in the proliferatively active mammary gland and that the rapidly dividing mammary epithelial cells are not susceptible to the action of cFABP during early pregnancy. © 1995 Wiley-Liss, Inc.
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  • 105
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 223-232 
    ISSN: 0192-253X
    Keywords: Genomic imprinting ; parthenogenetic embryos ; biallelic expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Uniparental embryos have been instrumental in studying imprinting because contributions from the parental genomes can be determined unambiguously. In this study, we set out to identify imprinted genes showing differential expression between parthenogenetic and fertilized embryos during preimplantation and early postimplantation stages of development. We identified three genes-apolipoprotein E, pyruvate kinase-3, and protein phosphatase 1 gamma-that represent excellent candidates for imprinted genes, based on the results of the differential screen, their function in differentiation and the cell cycle, and their location within imprinted chromosomal regions. In addition, two novel genes expressed in trophoblast were identified, 1661 and RA81. These genes, together with four known imprinted genes, H19, Igf2r, Igf2, and Snrpn, showed evidence of expression from both parental alleles in early stage embryos, indicating a role for postfertilization processes in regulating imprinted gene function. © 1995 Wiley-Liss, Inc.
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  • 106
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 233-239 
    ISSN: 0192-253X
    Keywords: Genome imprinting ; zebrafish ; Danio rerio ; methylation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have examined the inheritance of a transgene locus in the zebrafish, Daniorerio and demonstrated that its methylation is af fected by the sex of the parent contributing the allele. This parent-of-origin effect on the zebrafish transgene appears to be identical to imprinting as seen in mammals except that in zebrafish, passage of the locus through a female tended to decreased its methylation, whereas passage through a male increased it. Methylation of the transgene in gametic tissues differed from somatic tissue with the locus being hypomethylated in sperm and hypermethylated in the unfertilized egg. The potential identification of imprinting in the zebrafish has important ramifications with respect to the evolution of the process as well as for understanding the role of imprinting in mammals. © 1995 Wiley-Liss, Inc.
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  • 107
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 272-279 
    ISSN: 0192-253X
    Keywords: Gene amplification ; hypotrichous ciliates ; macronuclear genes ; gene expression ; recognition factors ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In hypotrich ciliates, the entire silent chromosomal genome of the germinal nucleus (micronucleus) undergoes extensive DNA rearrangements that, during the development of the somatic nucleus (macronucleus) at the beginning a new cell life cycle, eventually result in the production of linear DNA molecules. These molecules represent functional genes, each one consisting of a central coding region flanked by two shorter regions, which apparently lack canonical elements for regulation of replication and transcription. These are amplified to thousands of copies in the “adult” macronucleus of the vegetative cell. We defined the extent of this amplification for allelic codominant genes which, in the macronucleus of Euplotes raikovi, encode polypeptide cell recognition factors (pheromones). This amplification was shown to be allele-specific. The copy numbers of genes coding for pheromones Er-1, Er-2, and Er-10 were determined to be 2.5 - 2.9 × 104, 0.9 - 1.2 × 104, 1.6 - 1.85 × 104 respectively, and these numbers did not appreciably vary during the vegetative cell proliferation. This differential amplification of pheromone genes was (i) independent of whether two genes coexisted in the same heterozy-gous cell or were separated in the corresponding homozygotes, and (ii) directly correlated with quantitative variations in mRNA synthesis and pheromone secretion. On the basis of these results, it is suggested that a mechanism of gene-specific amplification may be used by hypotrich ciliates to modulate gene expression. © 1995 Wiley-Liss, Inc.
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  • 108
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 17 (1995), S. 253-262 
    ISSN: 0192-253X
    Keywords: IGF2 ; IDDM2 ; IDDM ; immune function ; IGFs ; apoptosis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Parental genomic imprinting is the phenomenon in which the behavior of a gene is modified, depending on the sex of the transmitting parent [Peterson and Sapienza (1993): Annu Rev Genet 27:7-31]. Recent observations have revealed that the inheritance patterns, age-of-onset, severity, and etiology of certain human diseases can be explained by aberrations in the establishment or the maintenance of the imprint. Examples include the Prader-Willi, Angelman, and Beckwith-Wiedemann syndromes [Nicholls (1994): Am J Hum Genet 54:733-740], malignancy [Sapienza (1990): Biochim Biophys Acta 1072:51-61; Feinberg (1993): Nat Genet 4:110-113], and insulin-ependent diabetes mellitus (IDDM) [Julier et al. (1994) Nature 354:155-159; Bennett et al. (1995) Nat Genet 9:284-292]. We review the evidence that implicates an imprinted gene in the INS-IGF2 region of chromosome llp15 in the etiology of IDDM (referred to as the IDDM2 locus) and show that in human fetal pancreas, INS is not imprinted, thus providing an argument against INS as the candidate gene. We also examine imprinting effects on the expression of IGF2 in components of the human immune system believed to be important in IDDM and show imprinted expression in fetal thymus as early as 15 weeks gestation. We demonstrate further that in the circulating mononuclear cells of two individuals, lectin-stimulated IGF2 transcription was biallelic, indicating relaxation of imprinting, whereas in one individual, transcription was monoallelic. Finally, we review the current available data supporting a role for insulin-like growth factor-ll (IGF-II) in the immune system and, more specifically, discuss the evidence supporting a role for the IGFs in the prevention of apoptosis. These data have led us to formulate a novel hypothesis that could mechanistically explain the involvement of the IDDM2 locus in the pathogenesis of IDDM. © 1995 Wiley-Liss, Inc.
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  • 109
    ISSN: 0192-253X
    Keywords: Translation elongation factor 1α (EF-1α) ; developmental regulation ; oogenesis ; microinjection ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Transcription of the Xenopus laevis EF-1αS gene commences at the mid-blastula stage of embryonic development and then continues constitutively in all somatic tissues. The EF-1αS promoter is extremely active in the early Xenopus embryo where EF-1αS transcripts account for as much as 40% of all new polyadenylated transcripts. We have isolated the Xenopus EF-1αS gene and used microinjection techniques to identify promoter elements responsible for embryonic transcription. These in vivo expression studies have identified an enhancer fragment, located approximately 4.4 kb upstream of the transcription start site, that is required for maximum expression from the EF-1αS promoter. The enhancer fragment contains both an octamer and a G/C box sequence, but mutation studies indicate that the octamer plays no significant role in regulation of EF-1αS expression in the embryo. The presence of a G/C element in the enhancer and of multiple G/C boxes in the proximal promoter region suggests that the G/C box binding protein, Spl, plays a major role in the developmental regulation of EF-1αS promoter activity. © 1995 Wiley-Liss, Inc.
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  • 110
    ISSN: 0192-253X
    Keywords: Ecdysone receptor ; Choristoneura fumiferana cDNA cloning ; developmental expression ; molting and metamorphosis ; ecdysone response element ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Degenerate oligonucleotides were designed on the basis of conserved amino acid sequences in the DNA and ligand-binding regions of the members of the steroid hormone receptor superfamily. Using these oligonucleotides in RNA-PCR, a cDNA fragment was isolated from the spruce budworm, Choristoneura fumiferana. Comparison of the deduced amino acid sequence of this cDNA fragment with the members of the steroid hormone receptor superfamily suggested that this PCR fragment is a region of the ecdysone receptor from C. fumiferana. Using this cDNA fragment as a probe, 10 clones were isolated from a cDNA library that was constructed using the RNA from 4- and 5-day old embryos of C. fumiferana. Two cDNA clones (1.3 and 3 kb) that overlap and show amino acid identity with Drosophila melanogaster ecdysone receptor B-1 isoform (DmEcR) were characterized and sequenced. The longest open reading frame had 539 codons and covered the complete EcR coding region. The deduced amino acid sequence of this open reading frame had all five of the regions typical for a steroid hormone nuclear receptor. The C domain or DNA binding region showed the highest identity with EcR proteins from D. melanogaster, Chironomus tentans, Aedes aegypti, Manduca sexta and Bombyx mori. The A/B region, D domain or hinge region, E domain, or ligand binding region also showed significant amino acid similarity with the EcR proteins from the five insects mentioned above. The C. fumiferana ecdysteroid receptor (CfEcR) cDNA probe detected a 6.0-kb mRNA that was present throughout the development of C. fumiferana. The CfEcR mRNA increases in abundance at the time of the ecdysteroid peak during the molting phase in the embryonic, larval and pupal stages but remains low during the intermolt period. In the 6th instar larvae, the 6-kb CfEcR mRNA was detected in the epidermis, fat body, and midgut and maximum expression was observed during the prepupal peak of ecdysteroids in the hemolymph. CfEcR mRNA was induced in ecdysone treated CF-203 cells as well as in the epidermis and midgut of larvae that were fed the nonsteroidal ecdysteroid agonist, RH-5992. The induction occurred within an hour and reached maximum levels around 3 hr, after which it decreased to the basal level by 6 hr. In vitro transcription and translation of the CfEcR cDNA yielded a 67-Kda protein that bound to the ecdysone response element (EcRE) as a heterodimer, along with the ultraspiracle protein.
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  • 111
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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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  • 112
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 75-75 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 113
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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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  • 114
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    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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  • 115
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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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  • 116
    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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  • 117
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    Developmental Genetics 6 (1985), S. 151-151 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 118
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    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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  • 119
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    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
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  • 120
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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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  • 121
    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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  • 122
    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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  • 123
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    Developmental Genetics 6 (1985), S. 199-212 
    ISSN: 0192-253X
    Keywords: ultrastructure ; cell death ; Drosophila melanogaster ; imaginal disk ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The temperature-sensitive mutant l(3)c43hs1 is lethal at the restrictive temperature late in the last larval instar and has wing disks that show excessive growth when larvae are reared at 25°C. Such mutant disks give rise to defective wings showing duplications and deficiencies. Abnormal folding patterns are localized to the region between the wing pouch and the area where adepithelial cells are found; the disks retain an epithelial morphology. Apoptotic cell death is distributed throughout the wing disks without any obvious concentration of dead cells in a specific area. Cell death is seen as early as 12 hr after a shift to the restrictive temperature. Temperature shift experiments also show that cell death precedes the onset of overgrowth, but since the spatial distribution of death is not localized to the regions of abnormal folds, it is unlikely that cell death and overgrowth are causally related.
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    Developmental Genetics 6 (1985), S. 179-197 
    ISSN: 0192-253X
    Keywords: embryonic development ; phenocopies ; heat shock ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Heat shock of pre-adult Drosophila disrupts development and causes phenotypic abnormalities. Type of abnormality depends on developmental stage at time of shock. Defects probably result from disruption of stage specific processes by the heat shock response (which includes reduction of normal mRNA and protein production). This study uses heat shock to study stage specific processes in early development. Short, intense shocks (2-3 min, 42-43°C) are administered to carefully staged embryos within the first 5 h of development. Stage specific defects occur following shock at syncitial blastoderm or later. Abnormal segmentation follows shock at syncitial or cellular blastoderm. Segmentation is also disrupted by shocks 1 h after the onset of gastrulation, but not by shocks at the onset of gastrulation. Segmentation defects include phenocopies of pair rule mutants, which lack parts of alternate segments. Defective shortening of the germ band is common following shock at the onset of gastrulation. Germ band shortening normally occurs several hours after the time of shock; thus heat shock specifically affects control of a later developmental process. Development does not simply cease at the time of the distrupted process; rather a specific step in the developmental sequence is omitted or altered. Stage specific defects do not occur following pre-blastoderm shock. Pre-blastoderm eggs have few or no normal processes controlled by transcription, and poor ability to induce the heat shock response. This suggests that stage specific defects require disruption of transcription controlled processes. Pre-blastoderm eggs survive a 3-min shock less well than older eggs. The ability of older eggs to induce the heat shock response probably enhances survival. The mutant hairy was also investigated. Extreme alleles show a striking pair rule phenotype, while a weak allele does not. Heat shock of animals heterozygous or homozygous for the weak allele at blastoderm specifically increases the frequency of the extreme hairy phenotype. Thus heat shock may disrupt the same developmental process as is altered by the mutation.
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    Developmental Genetics 6 (1985) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 126
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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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    Developmental Genetics 6 (1985), S. 239-246 
    ISSN: 0192-253X
    Keywords: Drosophila melanogaster ; germ line ; somatic line ; pole cell transplantation ; mosaics ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Drosophila melanogaster mutant fs(1)1304 is an ovary autonomous female sterile mutant that causes abnormal morphology of the egg. Vitellogenesis proceeds at an abnormally slow rate in homozygous females. We have used pole cell transplantation to construct germ line mosaics in order to determine whether the 1304 defect depends upon the genotype of the germ line cells (oocyte or nurse cells) or the somatic line (follicle cells). We have found that the germ line is the primary target tissue where the mutant gene is expressed.
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    Developmental Genetics 6 (1985), S. 295-296 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 6 (1985), S. 269-280 
    ISSN: 0192-253X
    Keywords: UV ; DNA repair ; photoreactivation ; algae ; dark repair ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The response of Volvox to ultraviolet irradiation was analyzed. Young individuals isolated from a synchronous culture were exposed to UV light (120 J/m2) and subjected to variable lenght periods of dark following irradiation. The major effect of the UV treatment was the inability of the gonidia present in the colonies at the time of irradiation to continue and complete the developmental program. Individuals show a heightened sensitivity to UV for a limited period immediately following inversion and are insensitive at other stages of development. The cytotoxic effect of UV during this interval is completely reversed by the immediate exposure to white light and is increased with longer periods of dark treatment prior to exposure to white light. The temporal profile of the sensitivity defines a smooth curve in which the maximal sensitivity occurs three hours after inversion. The response to higher doses of UV (up to 500 J/m2) is a nonlinear increase in cytotoxicity and is disproportionanately greater in those individuals just prior to the period of maximal sensitivity than those later in development. The results suggest that Volvox has at least two pathways for the repair of UV damage and that one of these, the principal dark repair pathway, is temporarily deficient in the gonidia of young individuals.
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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 6 (1985), S. 297-297 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 132
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    Developmental Genetics 7 (1986) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 7 (1986), S. 65-73 
    ISSN: 0192-253X
    Keywords: long interspersed repeated DNA ; demethylation ; myeloma cells ; aging ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sequences of DNA that hybridize on Southern blots with cloned EcoR1 1.3 kb (ER1) of long interspersed repeated sequence (L1Md) of mouse have been examined in genomic DNA of neonatal mice, livers and brains of adult mice (3, 10, 27, and 30 mo old), and the solid myeloma tumor MOPC-315. The isoschizomers Hpa II (CCGG or mCCGG) and Msp I (CCGG or CmCGG) were used to assess methylation. We found that the L1Md sequence is fully methylated in young animals but demethylated in myeloma. Demethylation of L1Md sequence also occurred in aged animals. By scanning the autoradiogram, we found that approximately 8% of the 104-105 copies have been demethylated in 27-mo-old liver.
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    Developmental Genetics 7 (1986), S. 75-80 
    ISSN: 0192-253X
    Keywords: DNA replication ; eggshell ; female sterile mutant gene amplification ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: K575 is a temperature-sensitive female sterile mutant which shows abnormal chorion structure and subnormal amounts of the major chorion proteins at the restrictive temperature. These phenotypes apparently result from a temperature-sensitive defect in amplification. Both clusters of chorion genes are affected, indicating that the gene operates in trans.
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    Developmental Genetics 7 (1986), S. 117-117 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 8 (1987), S. 1-9 
    ISSN: 0192-253X
    Keywords: T/t-complex ; LT mice ; parthenogenesis ; recombination ; gene-mapping ; primitive streak ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ovarian teratomas that result from parthenogenetic activation of oocytes provide a double tool for developmental geneties. First, they provide a way of measuring recombination between a gene and its centromere. Second, in the absence of crossing over there is the potential of producing tumors that are homozygous for genes that would be lethal in the course of in utero embryonic development. We have applied both aspects to several t- haplotypes containing different early acting t-lethal genes. In a study of 26 tumors, genotyped by Southern blot analysis of the major histocompatibility complex (MHC), we measured the distance between the centromere and the start of the t-complex as 5.6 ± 2.3 cM. We found a marked deficiency of t-homozygous genotypes among the tumors we studied, although T/T genotypes formed teratomas at levels comparable to controls. None of the lethal t-haplotypes we studied permit homozygous embryos to develop to the primitive streak stage, while T/T embryos do develop essentially normally through that stage. Thus, although the total number of tumors observed from t-bearing mice was small, the great difference in the incidence of t/t tumors versus the incidence of T/T tumors suggests strongly that the parthenogeretic embryos that convert to teratocarcinomas must first pass through some of the stages of normal early development, including the formation of three germ layers and the primitive streak.
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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. 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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    Developmental Genetics 8 (1987), S. 187-187 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 8 (1987) 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 8 (1987), S. i 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 9 (1988), S. 549-559 
    ISSN: 0192-253X
    Keywords: adhesion proteins ; development ; mutations ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three separate mechanisms of cell-cell adhesion have been shown to appear at different stages of development in Dictyostelium discoideum. During the first few hours of development, the cells synthesize and accumulate a glycoprotein of 24,000 daltons (gp24) that is positioned in the membrane. The time of appearance of gp24 correlates exactly with the time of appearance of cell-cell adhesion in two strains in which temporal control varies by several hours. Antibodies specific to gp24 are able to block cell-cell adhesion during the first few hours of development but not during later development. By 8 hr of development, another glycoprotein, gp80, that is not recognized by antibodies to gp24 accumulates on the surface of cells. This membrane protein mediates an independent adhesion mechanism during the aggregation stage that is resistant to 10 mM EDTA. Antibodies specific to gp80 can block EDTA-resistant adhesion during this stage. During subsequent development, gp80 is removed from the cell surface and replaced by another adhesion mechanism that is insensitive to antibodies to either gp24 or gp80.A λgtll expression vector carrying a Dictyostelium cDNA insert was isolated that directs the synthesis of a fusion protein recognized by antibodies specific to gp24. This cDNA was used to probe a genomic library. A clone carrying a 1.4-kb insert of genomic DNA was recognized by the cDNA and shown to hybridize to a 0.7-kb mRNA that accumulates early in development. This unusually small RNA could code for the small protein, gp24. Southern analysis of restriction fragments generated by various enzymes on Dictyostelium DNA with both the cDNA and genomic clones indicated the presence of two tandem copies of the gene. This may account for the failure to recover mutations resulting in the lack of gp24.Mutations have been recovered that result in the lack of accumulation of gp80, and cells carrying these mutations have been shown to be missing the second adhesion mechanism. These mutant strains are able to complete development because the other adhesion mechanisms are not impaired. Sequential addition of adhesion mechanisms provides a means for the formation of multicellular organisms from previously solitary cells.
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    Developmental Genetics 10 (1989), S. 63-69 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 10 (1989), S. 53-62 
    ISSN: 0192-253X
    Keywords: Mouse T-locus ; Two-dimensional gel electrophoresis ; Embryonic development ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: T and Tc are dominant mutations in the mouse that affect neuroaxial development when heterozygous and cause embryonic death when homozygous. Embryos were analyzed individually by two-dimensional gel electrophoresis at 9½ days gestation, 1 day before homozygotes die in utero. A comparison of the protein patterns of mutant homozygotes with those of their littermates revealed a set of proteins (T-proteins) that showed isoelectric point (pl) polymorphism. All the T-proteins were more basic in mutant homozygotes. These polymorphisms could be detected, although they were less pronounced, in embryos as young as 7½-day presomite stages, when it is impossible to distinguish homozygous mutants grossly. Interestingly, the same proteins show a pl shift from basic to acidic in wild-type embryos during development from 7½ to 9½ days. Thus, it appears that in T and Tc mutants a developmentally specific posttranslational acidic modification of these proteins is disturbed. The likely cause of the abnormality is a defect in some mechanism for phosphorylation, since the T-proteins of wild-type embryos were shifted to higher pls by phosphatase treatment. This disturbance appears to be localized to axial structures (neural tube, somites, and surrounding mesenchyme) since only these structures, and not the rest of the mutant homozygous embryos, contain abnormally basic T-proteins.
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    Developmental Genetics 10 (1989), S. 106-111 
    ISSN: 0192-253X
    Keywords: Bithorax complex ; Variable penetrance suppression ; Enhancement ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Two modifiers of bithorax1 phenotypic expression are described. An X-chromosome region is associated with sexual dimorphism in bx1 penetrance. It is hypothesized that sexual dimorphism is in part due to a lack of dosage compensation of the modifier, in males. A third chromosome region that segregates with the pink peach allele is implicated in mediating temperature sensitivity. By appropriate combinations of modifiers, both sexual dimorphism and temperature sensitivity can be greatly reduced.
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    Developmental Genetics 10 (1989) 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 10 (1989), S. 123-123 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 10 (1989), S. 210-219 
    ISSN: 0192-253X
    Keywords: Intron ; Alcohol dehydrogenase ; Enhancer ; Promoter ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have used in vitro mutagenesis and somatic transformation [Sofer and Martin, 1987a; Martin et al., 1986] to investigate the role of cis-acting sequences in the control of alcohol dehydrogenase gene expression in larvae of Drosophila melanogaster. Two sets of experiments were carried out. In the first, a series of aeletions were constructed in the region upstream of the proximal transcriptional start site. In the second, one or both introns were removed from within the structural gene. These constructs (on circular plasmids) were injected into Adh-null embryos and ADH activity was assayed in third instar larvae of the injected generation. The first set of experiments indicated that there are at least three distinct regulatory regions essential for larval activity located in the 5′ flanking region of the gene. One, in an area that includes the TATA box, was found to be necessary but not sufficient for larval ADH activity. Two others, further upstream, seemed to have enhancer-like properties because their absence could be compensated by a second copy of the Adh gene on the same plasmid molecule. The second set of experiments showed that neither the tis-sue distribution nor amount of ADH activity was affected by the removal of one or both introns from the Adh gene.
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    Developmental Genetics 10 (1989), S. 220-231 
    ISSN: 0192-253X
    Keywords: Drosophila ; Fat body ; Ecdysone ; cis-acting regulatory elements ; Development ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The P1 gene, together with the LSP-1a, -1β, and -ly, LSP-2, and P6 genes, is expressed exclusively in the larval fat body of D. melanogaster during the third instar. In vivo mapping of the cis-acting regulatory sequences of the P1 gene was carried out using hybrid constructs with three different reporter genes and a combination of transient and germline transformation assays. This revealed that regulatory elements involved in the setting up of the temporal and spatial specificities of transcription of the P1 gene are located in a short DNA region immediately upstream of the mRNA transcription start. This region includes on element that behaves as a fat-body transcriptional enhancer and element(s) required for ecdysone inducibility of transcription of the P1 gene.
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    Developmental Genetics 10 (1989), S. 232-238 
    ISSN: 0192-253X
    Keywords: Transcriptional regulation ; Alternate splicing ; Neurotransmitters ; Learning ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The gene Ddc encodes two isoforms of the enzyme dopa decarboxylase in Drosophila. These gene products catalyze the final steps in the synthesis of the biogenic amines sero-tonin and dopamine. This article summarizes recent progress in understanding the tissue- and cell-specific regulation of Ddc, which occurs at both the transcription and alternate splicing levels. In addition, results that are pertinent to understanding the roles of biogenic amines in the neurophysiology of Drosophila are discussed.
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  • 151
    ISSN: 0192-253X
    Keywords: Transposable element ; Transcription factor ; Suppression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have used the yellow gene of Drosophila melanogaster as a model system in which to study the molecular mechanisms by which the gypsy retrotransposon causes mutant phenotypes that can be reversed by nonalleiic mutations at the suppressor of Hairy-wing locus. This gene encodes a 109,000 dalton protein that contains an acidic domain and 12 copies of the Zn finger motif, which are characteristic of some transcription factors and DNA binding proteins. The suppressible y2 allele is caused by the insertion of the gypsy element at -700 bp from the start of transcription of the Yellow gene, resulting in a phenotype characterized by mouth parts and denticle belts in the larvae, and by bristles in the adults, that show wildtype coloration, but mutant wings and body cuticle in the adult flies. This phenotype is the result of the interaction of gypsy sequences homologous to mammalian enhancers with tissue-specific yellow transcriptional regulatory elements located upstream from the gypsy insertion site and responsible for the expression of the yellow gene in the mutated tissues. This interaction is dependent on the binding of the su(Hw) protein to the specific gypsy sequences involved in the induction of the mutant phenotype.
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  • 152
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    Developmental Genetics 10 (1989), S. 249-260 
    ISSN: 0192-253X
    Keywords: Regulatory genes ; Pc group ; Drosophila embryogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A wide variety of gain of function mutations have been induced in the Posterior Sex Comb (Psc) - Aristapedioid (Arp) - Suppressor 2 of zests(Su(z)2) region of the second chromosome of Drosophila. This region contains at least three apparently related genes, two of which we have been studying. Psc1 has previously been used to identify Psc as a Pc group gene; however, it is a complex mutation with both gain and loss of function character. We report here that the Pc group character of Psc is not due to a gain of function and presumably reflects the function of the wild-type gene. We also provide evidence for a maternal function for Psc, as well as the neighboring Su(z)2 gene.Su(z)2 does not appear to be a Pc group gene as it does not act in a synergistic fashion with other PC group genes in promoting posteriorly directed transformations. However, we have found that mutations in Su(z)2 do interact in a variety of interesting ways with mutations in Pc group genes.
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  • 153
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    Developmental Genetics 10 (1989), S. 311-317 
    ISSN: 0192-253X
    Keywords: β-globin ; Human erythroleukemia cells ; RNA transcripts ; K562 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Previous studies have indicated that control and hemin-treated human eryth-roleukemia K-562 cells fail to produce adult-type β-globin mRNA transcripts and to translate them into nascent β-globin chains. Expression of the β-globin DNA sequences in K-562 cells can occur, however, under certain conditions. To readdress this issue and to examine the possibility of whether these cells produce immature and untranslatable β-globin RNA transcripts, we prepared total cyto-plasmic RNA from control and inducer-treated cells and performed Northern blot hybridization analysis using 5′ end-labeled fragments of the human β-globin DNA rather than 3′ end fragments as probes. Although hybridization of both cytoplasmic and nuclear K-562 RNA with a32P-labeled 3′ end fragment (1.6kb Bam H1 cut) coding for a large part of the first exon of β-globin failed to detect β-globin RNA transcripts, hybridization with a 5′ end 32P-labeled 2.0kb Bam H1 fragment (coding for the third exon and part of the second) revealed the presence of relatively small (〈7S) RNA molecules both in nuclear and cytoplasmic fraction. S1 nuclease mapping of both cytoplasmic and nuclear RNA with the use of 5′ end-labeled 2.0 kb Bam H1 fragment of human β-globin DNA indicated protection of a small portion located 64bp 5′ upstream from the Bam H1 site of the second exon. The amount of protected portion was relatively higher in K-562 cells undergoing erythroid maturation. These findings suggest that control and differentiating K-562 cells synthesize β-globin-like RNA transcripts that are 3′ end short, immature, and unable to give rise to adult β-globin chains. These results also indicate that K-562 cells may lack factors that are unique for transcription and processing of the human β-globin RNA transcripts.
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  • 154
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    Developmental Genetics 10 (1989), S. 345-345 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 155
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    Developmental Genetics 10 (1989) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 156
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    Developmental Genetics 10 (1989), S. 347-347 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 157
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    Developmental Genetics 10 (1989), S. 339-344 
    ISSN: 0192-253X
    Keywords: Delayed processing ; Splicing ; Transcription ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This study deals with the pattern of developmental expression of the catalase gene in mice. We have used a mouse catalase 2 kb cDNA (pMCT-1) and its 1.4 kb 5′ fragment as probes to characterize the transcripts during embryonic development and differentiation. Total RNA was isolated from 8 days postconceptus (p.c.) whole embryos and from livers and carcasses of 13, 15, and 18 day p.c. embryos as well as from the livers of newborn and adult mice of the S.W. strain. The RNA was applied on slot blots, and run on agarose gels to generate northern blots. Blots were hybridized with the 32P-labeled cDNA probe under different stringency conditions. Autoradiograms were scanned with a densitometer to quantify relative hybridization signals of RNA samples obtained from two or three individual mice representing each stage of development.The catalase transcript is detectable as early as 8 days p.c. with the beginning of somite formation. At this stage, it is primarily in the form of a 12.2 kb transcript. One additional band (2.4 kb) is also apparent at this stage although at a very low intensity. The intensity of the two bands increases with development, particularly during 13-18 days p.c. in liver and carcass. The 2.4 kb RNA band increases sharply from day 8 through 13, 15, and 18 days p.c. and is confined primarily to the liver. Interestingly, only the 2.4 kb RNA band is seen at and after birth. The 2.4 kb RNA is the known mature message of the catalase gene in mice. The presence of large catalase-specific RNA species (seen during development in utero only) is interpreted as the primary transcript of this gene. The complete and efficient processing of this primary transcript takes place only after birth and primarily in the liver, which may be related to the physiological role of this enzyme in oxygen metabolism, particularly stressful superoxides, expected with independent respiration. At a lower stringency wash of the northern blots, a 9.5 kb RNA was seen during a narrow window of in utero development. This 9.5 kb band may represent an uncharacterized catalase-related gene with a possible role in development and differentiation.
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  • 158
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    Developmental Genetics 10 (1989), S. 349-355 
    ISSN: 0192-253X
    Keywords: SV40 promoter ; Expression vector ; Drug resistance ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have previously demonstrated systemic resistance to methotrexate (MTX) in transgenic mice carrying a foreign, mutant dihydrofolate reductase (DHFR, E.C. 1.5.1.3) gene. The new gene was introduced as a cDNA cloned into an expression vector driven by the simian virus 40 (SV40) early promoter. Previous physiologic studies suggested that transgenic mice tolerated drug doses invariably lethal to controls on the basis of gastrointestinal (GI) resistance to MTX. In the present study we evaluated foreign gene expression at the RNA level in the three major sites of MTX toxicity: intestine, liver, and bone marrow.The transgene was transcriptionally active in small bowel, and levels of expression were high in animals tolerating the largest doses of MTX. The gene was also expressed in the liver in some pedigrees, but was not detected in hemopoietic tissues of any of the pedigrees tested. Our studies correlate the site of expression of a drug resistant dhfr gene with an altered physiologic response to MTX, and demonstrate that transgenic mice can be used as a test system for expression of genes considered for use in somatic gene therapy.
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  • 159
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    Developmental Genetics 10 (1989), S. 356-364 
    ISSN: 0192-253X
    Keywords: Glucose intolerance ; Insulin resistance ; Diabetes mellitus ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We are investigating human insulin gene expression in transgenic mice. An 8.8 kilobase (kb) human genomic DNA fragment, including the insulin gene (1.4 kb) and 2 kb of 5′ human flanking sequences, was introduced into mouse embryos by pronuclear microinjection. Two lines of transgenic mice have been established, both of which carry the intact human gene in multiple copies. Animals from both lines have significantly higher insulin levels than control mice, and the degree of hyperinsulinemia shows a positive correlation with human gene copy number in the two lines. Expression of the human gene is confirmed by the detection of human C-peptide in plasma. Tissue specificity of expression is maintained, with human insulin mRNA detectable only in the pancreas. The transgenics maintain normal fasting blood glucose in spite of their high insulin levels, but preliminary studies show them to be glucose intolerant when given a glucose load. These mice provide a model system for further studies on the regulation of insulin gene expression and on the effects of chronic hyperinsulinemia on glucose homeostasis.
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  • 160
    ISSN: 0192-253X
    Keywords: Pentraxins ; Acute phase protein ; Lipopolysaccharide ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: To analyze the regulation of human serum amyloid P component (SAP) gene expression, we have produced seven transgenic mice. The 3.3 kb human SAP genes containing about 0.8 kb of 5′ and 1.5 kb of 3′ flanking region were injected into fertilized eggs of C57BL/6 mice. In five of the seven transgenic mice, human SAP was detected in the sera and serum concentrations were higher than that of human serum in three lines. The human SAP gene was expressed only in the liver. Amounts of human mRNA in the liver and serum concentrations of human SAP were roughly proportional to the copy number of the integrated gene. Human SAP production lowered the serum levels of mouse endogenous SAP. With the intraperitoneal administration of lipopolysaccharide, the mRNA levels in the liver and serum levels of mouse SAP increased several-fold in both the control and transgenic mice. On the other hand, neither the mRNA nor the serum levels of human SAP increased significantly.
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  • 161
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    Developmental Genetics 10 (1989), S. 386-392 
    ISSN: 0192-253X
    Keywords: Vestigial ; Cell death ; Modifier genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An analysis of the modifiers affecting the expression of the vg gene was performed. We selected for weak and strong expression of the vg mutant in 2 segregating populations obtained by crossing a vestigial stock with an Oregon laboratory stock (O) and with a wild strain (B) captured near Bologna, Italy. The selection for enlarged wings was more effective in the vg B population where wild wings appeared from the lCth generation. The assay of the three major chromosomes showed that the modifiers are located on chromosomes 2 and 3. The mutant imaginal disc cell death phenotype is evident in vg/vg strains that have a wild-type wing phenotype. It is suggested that the selected modifiers do not prevent cell death but induce regenerative growth.
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  • 162
    ISSN: 0192-253X
    Keywords: Monoclonal antibodies ; Myogenesis ; Adult isoforms ; Quail ; Chicken ; Muscle development ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have raised monoclonal antibodies (Mabs) to myosin heavy chain isoforms (MHCs) that have specific patterns of temporal expression during the development of quail pectoral muscle and that are expressed in very restricted, tissue-specific patterns in adult birds. We find that an early embryonic, a perinatal, and an adult-specific, fast myosin heavy chain a.e co-expressed at different levels in the pectoral muscle of 8-12 day quail embryos. The early embryonic MHC disappears from the pectoral muscle at approximately 14 days in ovo, whereas the perinatal MHC persists until 26 days post-hatching. The adult-specific MHC accumulates preferentially and eventually completely replaces the other isoforms. These Mabs cross-react with the homologous isoforms of the chick and detect a similar pattern of MHC expression in the pectoral muscle of developing chicks. Although the early embryonic and perinatal MHC isoforms recognized by our Mabs are expressed in the pectoral muscle only during distinct developmental stages, our Mabs also recognize MHC isoforms present in the heart and extraocular muscle of adult quail. Immunofingerprinting using Staphylococcus aureus protease V8 suggests that the early embryonic and perinatal MHC isoforms that we see are strongly homologous with the adult ventricular and extraocular muscle isoforms, respectively. These observations suggest that at least three distinct MHC isoforms, which are normally expressed in adult muscles, are co-expressed during the early development of the pectoral muscle in birds. In this respect, the pattern of expression of the MHCs recognized by our Mabs in developing, fast muscle is very similar to the patterns described for other muscle contractile proteins.
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  • 163
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    Developmental Genetics 10 (1989) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 164
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    Developmental Genetics 10 (1989), S. 460-472 
    ISSN: 0192-253X
    Keywords: Transposable elements ; Maize ; Mutation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The high frequency of mutations in Mutator stocks of maize is the result of transposition of Mu elements. Nine different Mu elements that share the 220 bp Mu terminal inverted repeats have been described. Mul elements have been found inserted into most of the molecularly characterized mutant alleles isolated from Mutator stocks, and most Mutator stocks contain a high number of Mul elements (10-60). However, it is clear that additional Mu elements, which share the Mul termini but have unrelated internal sequences, can also transpose in Mutator stocks. We were interested in comparing the mutation frequency and type of elements that inserted into a particular locus when Mutator stocks with differing numbers of Mul elements were utilized. Furthermore, previous studies with Mu-induced mutations have demonstrated that the element that inserted most frequently was Mul. Therefore, to try to obtain Mu elements different from Mul we utilized a stock that had a low number (3-6) of Mul elements as well as a Mutator stock with a more typical number of Mul elements (20-60).Utilizing both stocks, we isolated numerous mutants at one gene, Bronze1 (Bz1), and compared the type of elements inserted. In this paper we report that both the high and low Mu1 stocks produced bz1 mutants at frequencies characteristic of Mutator stocks, 6.6 and 4.3 ± 10-5, respectively. We describe the isolation of 20 bz1 mutations, and the initial molecular characterization of eight unstable mutations: two from the high Mu1 stock and six from the low Mu1 stock. The six alleles isolated from the low Mu1 stock appear to contain deleted Mu1 elements, and the two alleles isolated from the high Mu1 stock contain elements very similar to Mu1. When the mutants from the low Mu1 stocks were examined, it was found that the Mu1 -related elements increased from 3-6 copies to 9-20 copies in one generation. The high number of Mu1 -related elements was maintained in subsequent out-crosses. This spontaneous activation and amplification of Mu -related elements occurred in at least 1% of the low Mu1 plants.
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  • 165
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    Developmental Genetics 10 (1989), S. 482-506 
    ISSN: 0192-253X
    Keywords: Mutator ; Transposable elements ; Controlling elements ; Autonomous elements ; Regulator elements ; Mutable genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Mutator transposable element system (Mu) of maize has been responsible for the induction of numerous mutable aleurone mutants of maize. Unlike similar mutants induced by other transposable element systems, the mutability of Mu-induced mutants did not seem initially to be regulated by an independent autonomous or regulator element. However, in a continuing study of two Mu-induced a1 mutable mutants (a1-Mum2) and a1-Mum3, lines have been obtained that give evidence of an independently segregating regulator of somatic mutability. Data from several generations of crossing are presented indicating that intense somatic mutability in many of these stocks is under the control of an independent regulator. However, testing of other lines, which initially gave evidence of the presence of an independent regulator, were negative. Some of these latter lines could be expected to have Mutator elements that were modified (methylated) at sites recognized by certain restriction endonucleases. Modification of Mu elements, which is known to affect the expression of somatic mutability, might, at times, be responsible for producing conditions that mimic the segregation of an independent regulator. Lines with stable derivatives of the a1-Mum2 and a1-Mum3 can recover intense somatic mutability by crossing with germinally active Mutator stocks. Thus, active Mutator lines contain regulator elements and evidence is presented suggesting that such lines have multiple copies of these elements. Most a1- Mum2 and a1-Mum3 stocks segregating for a regulator do not have germinal Mutator activity. Thus the presence of one or a few putative regulator elements does not necessarily account for the high level of germinal activity in most Mutator stocks.
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  • 166
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    Developmental Genetics 13 (1992), S. 387-391 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 167
    ISSN: 0192-253X
    Keywords: hsp70 ; heat shock ; fungus ; steroid hormone ; secretion ; mycelial branching ; sexual differentiation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the filamentous oomycete fungus Achlya, the differentiation of gamete bearing structures on vegetative hyphae of the male mating type, is induced by the Achlya steroid hormone, antheridiol. Among the several metabolically labeled intracellular proteins whose synthesis or accumulation is altered by hormone treatment are steroid-induced 85-kDa and 68- to 78-kDa proteins. The 85-kDa protein was previously shown to be the Achlya heat shock protein hsp85 [Brunt et al., 1990; Brunt and Silver, 1991], a component of the putative Achlya steroid hormone receptor. It was of interest to determine if the antheridiol-induced “70-kDa” proteins were hsp70-family heat shock proteins and if hormone treatment-induced changes in the level of hsp70 transcripts. Two different Achlya hsp70 genomic sequences were cloned and used to investigate these questions. The two hsp70 sequences recognized two different mycelial transcript populations, one of which was regulated also by decreased glucose. Of note, both of the two hsp70 transcript populations were found to be regulated by antheridiol. The hormone-induced chcnges in hsp70 transcript levels were temporally correlated with the onset of massive lateral hyphal branching and alterations in the pattern of secreted N-linked glycoproteins which occur in hormone-treated mycelia. To our kncwledge, this represents one of the first reports on changes in hsp70 proteins and transcripts during fungal differentiation. Our results may have implications for the role of heat shock proteins in hyphal branching and secretion in filamentous fungi and perhaps other cell types. © 1993Wiley-Liss, Inc. Inc.
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  • 168
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    Developmental Genetics 14 (1993), S. 58-68 
    ISSN: 0192-253X
    Keywords: Ferritin ; heat shock ; development ; sea urchin ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 20-kD protein identified as a subunit of the iron-binding protein ferritin is present in S. purpuratus and L. pictus sea urchin embryos. The synthesis of the protein is stimulated by an elevation in temperature or by an increase in iron supply. The developmental expression of this protein and its regulation during normal development and upon heat shock was investigated. In L. pictus, ferritin is present in the unfertilized egg and, as determined by Western blot analysis, its concentration remains approximately constant after fertilization up to the gastrulc-pluteus stage; there is a small transient decrease in the level of the protein in the early blastula at a time coinciding with the first clear indication of its de novo synthesis. Northern blots reveal no cytoplasmic ferritin transcripts in the unfertilized egg, but there occurs a dramatic increase in the RNA level from the late morulaearly blastula stage (12-14 hr) to the mesenchyme blastula-early gastrula (25-30 hr) stage. This developmentally regulated increase in the constitutive concentration of ferritin RNA is correlatable with the normal onset of synthesis of the protein. The overall degree and nature of induction of ferritin by heat is dependent on the developmental stage: at 10-16 hr postfertilization heat shock elicits an increase in both the concentration of RNA and the synthesis of the protein; in hatched blastula (18 hr) and in later embryos heat shock increases ferritin synthesis, without a corresponding increase in the mRNA level. It appears that different mechanisms operate in the developing sea urchin embryo to regulate the expression of ferritin during normal development and on exposure to heat stress, one dependent on the concentration of ferritin transcripts and another operating at the level of translational control. © 1993Wiley-Liss, Inc.
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    Developmental Genetics 14 (1993), S. 78-86 
    ISSN: 0192-253X
    Keywords: Heat shock ; stress ; senescence ; Drosophila ; ubiquitin ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We examined the effect of aging on the expression of ubiquitin RNA and the binding of the ubiquitin polypeptide to proteins following heat shock in Drosophila melanogaster. Heat-shocked adult flies transcribe two major RNA species-one of 4.4 kb and one of about 6 kb that hybridize to the polyubiquitin-encoding probe. Several less abundant RNAs were also observed but the 4.4-kb band was present as the major RNA species in both stressed and nonstressed flies of both ages. The 6-kb fragment was more abundant in heat shocked aged flies than in younger flies. The quantitative expression of the polyubiquitin gene increased in proportion to the duration of the heat stress. Moreover, the induction of the polyubiquitin RNA was markedly elevated during aging following heat shock. Hybridization of Northern blots with the monoubiquitin gene probe revealed a band of 0.9 kb that was not significantly affected by heat stress.We also investigated the relationship between the changes in polyubiquitin gene expression and the formation of ubiquitin-protein complexes in aging heat-shocked flies. Heat shock to old flies results in a significant increase in the level of proteins immunoprecipitated by anti-ubiquitin antibodies. In the case of proteins synthesized 2 h before heat shock, most of the ubiquitinated proteins were of high molecular weight. For those proteins synthesized during a 30-min heat shock and the 2 h following heat shock, two major immunoprecipitated bands were observed: an 80-kD and a 70-kD polypeptide. The ubiquitination of a 60 kD protein was also observed in nonstressed flies, but its for mation was drastically reduced following heat shock. For proteins synthesized during and after heat shock from both age groups, the major ubiquitinated polypeptide is 70 kD. In all age groups, more ubiquitin complexes were formed with proteins synthesized before heat shock, than with proteins synthesized either during or after heat shock. This suggests that cellular proteins synthesized at physiological temperatures are more sensitive to heat induced damage than those synthesized during stress. These data support the hypothesis that in aging flies, heat shock induces an unusually high concentration of abnormal proteins which are targeted for degradation by the ubiquitin-dependent proteolytic system. © 1993Wiley-Liss, Inc.
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  • 170
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    Developmental Genetics 14 (1993), S. 119-126 
    ISSN: 0192-253X
    Keywords: Spermatogenesis ; HSP90 proteins ; HSP70 proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This study extends to the protein level our previous observations, which had established the stage and cellular specificity of expression of hsp86 and hsp84 in the murine testis in the absence of exogenous stress. Immunoblot analysis was used to demonstrate that HSP86 protein was present throughout testicular development and that its levels increased with the appearance of differentiating germ cells. HSP86 was most abundant in the germ cell population and was present at significantly lower levels in the somatic cells. By contrast, the HSP84 protein was detected in the somatic cells of the testis rather than in germ cells. The steady-state levels of HSP86 and HSP84 paralleled the pattern of the expression of their respective mRNAs, suggesting that regulation at the level of translation was not a major mechanism controlling hsp90 gene expression in testicular cells. Immunoprecipitation analysis revealed that a 70-kDa protein coprecipitated with the HSP86/HSP84 proteins in testicular homogenates. This protein was identified as an HSP70 family member by immunoblot analysis, suggesting that HSP70 and HSP90 family members interact in testicular cells. © 1993Wiley-Liss, Inc.
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  • 171
    ISSN: 0192-253X
    Keywords: Heat-shock expression ; cell cycle ; embryogenesis ; thermotolerance ; teratogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Synchronized regulation of cell division during gastrulation is essential for the regional proliferation of cells and pattern formation of the early CNS. The neural plate and neuroectoderm cells are a rapidly dividing and differentiating population of cells with a unique and rapid heat-shock response. Heat shock and the heat-shock genes were studied during neural plate development in a whole rat embryo culture system at 9.5-11.5 days. A lethal heat shock can cause cell death and severe developmental defects to the forebrain and eye during organogenesis. Heat shock can also result in acquired thermotolerance whereby cell progression is delayed at the G1/S and S/G2 boundaries of the cell cycle. This delay in cell cycle progression caused an overall lengthening of the cell cycle time of at least 2 hr. The heat shock genes may therefore function as cell cycle regulators in neuroectoderm induction and differentiation. The kinetics and expression of the hsp genes were examined in neuroectodermal cells by flow cytometry and Northern analysis. The levels of hsp mRNA 27, 71, 73, and 88 were identified following exposure at 42°C (nonlethal), 43deg;C (lethal) and 42deg;/43deg;C (thermotolerant) heat shock. Examination of hsp gene expression in the neural plate showed tight regulation in the cell cycle phases. Hsp 88 expression was enhanced at Go and hsp71 induction at G2 + M of the cell cycle. Cells exposed to a thermotolerant heat shock of 42deg;C induced hsp71 mRNA expression in all phases of the cell cycle with the mRNA levels of hsp27, 73, and 88 increased but relatively constant. Following a lethal heat shock, dramatic changes in hsp expression were seen especially enhanced hsp71 induction in late S phase. The regulated expression of hsps during the cell cycle at various phases could play a unique and important role in the fate and recovery of neuroectoderm cells during early mammalian embryo development. © 1993Wiley-Liss, Inc.
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  • 172
    ISSN: 0192-253X
    Keywords: Antisense inhibition ; Wnt-1 ; Wnt-3a ; Neural crest ; Central nervous system ; Hindbrain ; Midbrain ; Spinal cord ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Wnt-1 and Wnt-3a proto-on-cogenes have been implicated in the development of midbrain and hindbrain structures. Evidence for such a role has been derived from in situ hybridization studies showing Wnt-1 and -3a expression in developing cranial and spinal cord regions and from studies of mutant mice whose Wnt-1 genes have undergone targeted disruption by homologous recombination. Wnt-1 null mutants exhibit cranial defects but no spinal cord abnormalities, despite expression of the gene in these regions. The absence of spinal cord abnormalities is thought to be due to a functional compensation of the Wnt-1 deficiency by related genes, a problem that has complicated the analysis of null mutants of other developmental genes as well. Herein, we describe the attenuation of Wnt-1 expression using antisense oligonucleotide inhibition in mouse embryos grown in culture. We induce similar mid- and hindbrain abnormalities as those seen in the Wnt-1 null mutant mice. Attentuation of Wnt-1 expression was also associated with cardiomegaly resulting in hemostasis. These findings are consistent with the possibility that a subset of Wnt-1 expressing cells include neural crest cells known to contribute to septation of the truncus arteriosus and to formation of the visceral arches. Antisense knockout of Wnt-3a, a gene structurely related to Wnt-1, targeted the forebrain and midbrain region, which were hy-poplastic and failed to expand, and the spinal cord, which exhibited lateral outpocketings at the level of the forelimb buds. Dual antisense knockouts of Wnt-1 and Wnt-3a targeted all brain regions leading to incomplete closure of the cranial neural folds, and an increase in the number and severity of outpocketings along the spinal cord, suggesting that these genes complement one another to produce normal patterning of the spinal cord. The short time required to assess the mutant phenotype (2 days) and the need for limited sequence information of the target gene (20-25 nu-cleotides) make this antisense oligonucleotide/ whole embryo culture system ideal for testing the importance of specific genes and their interactions in murine embryonic development. © 1993 Wiley-Liss, Inc.
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    Developmental Genetics 15 (1994), S. 51-63 
    ISSN: 0192-253X
    Keywords: Actin ; ascidian development ; gene expression ; heterochrony ; muscle actin gene ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Adultation is a hetercchronic mode of development in which adult tissues and organs differentiate precociously during the larval phase. We have investigated the expression of an adult muscle actin gene during adultation in the ascidian Molgula citrina. Ascidians contain multiple muscle actin genes which are expressed in the larva, the adult, or during both phases of the life cycle. In ascidian species with conventional larval development, the larval mesenchyme cells, which are believed to be progenitors of the adult mesoderm, remain undifferentiated and do not express the muscle actin genes. In M. citrina, the mesen-chyme cells differentiate precociously during larval development, suggesting a role in adultation. An adult muscle actin gene from M. citrina was obtained by screening a mantle cDNA library with a probe containing the coding region of SpMAl, a Styela plicata adult muscle actin gene. The screen yielded a cDNA clone designated McMAl, which contained virtually the complete coding and 3′ noncoding regions of a muscle actin gene. The deduced McMAl and SpMAl proteins exhibit 97% identity in amino acid sequence and may be encoded by homologous genes. The McMAl gene is expressed in juveniles and adults, but not in larval tail muscle cells, suggesting that it is an adult muscle actin gene. In situ hybridization with a 3′ non-coding region probe was used to determine whether the McMAl gene is expressed during adultation in M. citrina. McMAl mRNA was first detected exclusively in the mesenchyme cells during the late tailbud stage and continued to accumulate in these cells during their migration into the future body cavity and heart primordium in the hatched larva. The McMAl transcripts persisted in mesenchyme cells after larval metamorphosis. It is concluded that an adult muscle actin gene shows a heterochronic shift of expression into the larval phase during adultation in M. citrina.
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  • 174
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    Developmental Genetics 15 (1994) 
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    Keywords: Life and Medical Sciences ; Genetics
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    Developmental Genetics 15 (1994), S. 176-187 
    ISSN: 0192-253X
    Keywords: Floral development ; floral genetics ; Tunicate maize ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The co-dominant Tunicate (Tu) mutation in maize causes nonreproductive structures in both the male and female inflorescences to be enlarged. This mutation also affects sex determination, permitting the development of pistils in the normally staminate tassel. In order to characterize the role of the normal tu gene product, we have analysed genetic interaction between Tu and other mutations that perturb specific stages of floral development. Synergistic interactions observed suggested that the tu product functions in at least three stages of floral development-determination of spikelet primordia, differentiation of non-reproductive organs and pistil abortion in the tassel. © 1994 Wiley-Liss, Inc.
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  • 176
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    Developmental Genetics 15 (1994), S. 205-213 
    ISSN: 0192-253X
    Keywords: Sex ; sexual reproduction ; recombination ; diploidy ; anisogamy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Despite the obvious efficiencies of many forms of asexual reproduction, sexual reproduction abounds. Asexual species, for the most part, are relatively short-lived offshoots of sexual ancestors. From the nineteenth century, it has been recognized that, since there is no obvious advantage to the individuals involved, the advantages of sexual reproduction must be evolutionary. Furthermore, the advantage must be substantial; for example, producing males entails a two-fold cost, compared to dispensing with them and reproducing by parthenogenetic females.There are a large number of plausible hypotheses. To me the most convincing of these are two. The first hypothesis, and the oldest, is that sexual reproduction offers the opportunity to produce recombinant types that can make the population better able to keep up with changes in the environment. Although the subject of a great deal of work, and despite its great plausibility, the hypothesis has been very difficult to test by critical observations or experiments.Second, species with recombination can bunch harmful mutations together and eliminate several in a single “genetic death.” Asexual species, can eliminate them only in the same genotype in which they occurred. If the rate of occurrence of deleterious mutations is one or more per zygote, some mechanism for eliminating them efficiently must exist. A test of this mutation load hypothesis for sexual reproduction, then, is to find whether deleterious mutation rates in general are this high-as Drosophila data argue. Unfortunately, although molecular and evolutionary studies can give information on the total mutation rate, they cannot determine what fraction are deleterious.In addition, there are short discussions of the advantages of diploidy, anisogamy, and separate sexes. © 1994 Wiley-Liss, Inc.
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  • 177
    ISSN: 0192-253X
    Keywords: Sex determination ; sexual differentiation ; reptiles ; temperature-dependent sex determination ; behavior ; steroidogenic enzymes ; aromatase ; reductase ; estrogen ; androgen ; steroid hormone receptors ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In many egg-laying reptiles, the incubation temperature of the egg determines the sex of the offspring, a process known as temperature-dependent sex determination (TSD). In TSD sex determination is an “all or none” process and intersexes are rarely formed. How is the external signal of temperature transduced into a genetic signal that determines gonadal sex and channels sexual development? Studies with the red-eared slider turtle have focused on the physiological, biochemical, and molecular cascades initiated by the temperature signal. Both male and female development are active processes - rather than the crganized/default system characteristic of vertebrates with genotypic sex determination - that require simultaneous activation and suppression of testis- and ovary-determining cascades for normal sex determination. It appears that temperature accomplishes this end by acting on genes encoaing for steroidogenic enzymes and steroid hormone receptors and modifying the endocrine microenvironment in the embryo. The temperature experienced in development also has long-term functional outcomes in addition to sex determination. Research with the leopard gecko indicates that incubation temperature as well as steroid hormones serve as organizers in shaping the adult phenotype, with temperature modulating sex hormone action in sexual differentiation. Finally, practical applications of this research have emerged for the conservation and restoration of endangered egg-laying reptiles as well as the embryonic development of reptiles as biomarkers to monitor the estrogenic effects of common environmental contaminants. © 1994 Wiley-Liss, Inc.
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    Developmental Genetics 16 (1995), S. i 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 179
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    Developmental Genetics 15 (1994), S. 523-532 
    ISSN: 0192-253X
    Keywords: Epigenetic ; paramutation ; cosuppression ; pattern elaboration ; flower pigmentation ; plant morphogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Developmental and physiological factors can impose heritable metastable changes on the plant genome, a fact that was established by maize geneticists during the 1950s and 1960s, largely through the efforts of R. Alexander Brink and Barbara McClintock. This paper describes a transgenic reporter system that monitors genomic impositions as changes in morphogenetically-determined flower color patterns. The observations reported here on the metastable properties of plant transgenes illustrate the proposals of Brink and McClintock that chromosomal impositions occur during normal development as ordered sequences of events which contribute to the elaboration of complex developmental patterns. The relationship between this process and some recent findings about the control of gene expression in transgenic plants is also discussed. © 1994 Wiley-Liss, Inc.
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    Developmental Genetics 16 (1995), S. 13-19 
    ISSN: 0192-253X
    Keywords: Meiotic maturation ; chromatin ; centrosomes ; microtubules ; oocytes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Meiotic maturation progresses atypically in oocytes of strain LT/Sv and l/LnJ mice. LT/Sv occytes show a high frequency of metaphase l-arrest and parthenogenetic activation. l/LnJ oocytes display retarded kinetics of meiotic maturation and a high frequency of metaphase l-arrest. Some l/LnJ oocytes fail to resume meiosis. Changes in the configuration of chromatin, microtubules, and centrosomes are associated with specific stages of meiotic progression. In this study, the configuration of these subcellular components was examined in LT/Sv, l/LnJ, and C57BL/6J (control) oocytes either freshly isolated from large antral follicles or after culture for 15 hr to allow progression of spontaneous meiotic maturation. Differences were found in the organization of chromatin, microtubules, and centrosomes in LT/Sv and l/LnJ oocytes compared to control oocytes. For example, rather than exhibiting multiple cytoplasmic and nuclear centrosomes as in the normal germinal vesicle-stage oocytes, LT/Sv oocytes typically contain a single large centrosome. In contrast, l/LnJ oocytes displayed many small centrosomes. The microtubules of normal germinal vesicle-stage oocytes were organized as arrays or asters, but microtubules were shorter in LT/Sv oocytes and absent from l/LnJ oocytes. After a 15-hr culture, centrosomal material of normal metaphase II oocytes was organized at both spindle poles. In contrast, metaphase l-arrested LT/Sv oocytes exhibited an elongated spindle with centrosomal material appearing more organized at one pole of the spindle. Both control and LT/Sv oocytes displayed cytoplasmic centrosomes. Metaphase l-arrested l/LnJ oocytes rarely had cytoplasmic centrosomes but exhibited centrosomal foci at the spindle periphery. Thus, oocytes that are atypical in the progression of meiotic maturation displayed aberrant configurations of microtubules and centrosomes, which are thought to participate in the regulation of meiotic maturation.
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    Developmental Genetics 16 (1995), S. 77-84 
    ISSN: 0192-253X
    Keywords: Ecdysone ; Dopa decarboxylase ; Drosophila melanogaster ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The quontitation of RNA in tissue homogenates by amplifying the product of reverse transcription (RT-PCR) is sufficiently sensitive to detect molecules in the range of 10-110-2 amole. We describe here the steps we believe necessary to validate a protocal that used a DNA competitor and visualization of the amplification products by ethidium bromide staining. The procedure was designed to quantitate one of the tissue specific transcripts of the Dopa decarboxylase gene (Ddc) in Drosophila. We demonstrate that the amount of epidermal Ddc transcript is much lower at pupariation in several mutants of the Broad-Complex, one of the primary response loci of the moulting hormone, ecdysone. The mutant effects were allele specific and the molecular basis of one of these alleles is known. This implicates a particular family of the zinc finger proteins encoded by the locus in the hormone dependent induction of Ddc expression.
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  • 182
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    Developmental Genetics 16 (1995), S. 140-147 
    ISSN: 0192-253X
    Keywords: Programmed cell death ; apoptosis ; spermatogenesis ; premeiotic stages ; testis ; in vitro regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Control points of regulator action during spermatogenesis are not completely known. Using the shark testis model, which facilitates analysis of spermatogenesis stage-by-stage in vivo and in vitro, an early biochemical marker of programmed cell death (PCD) was detected. Nucleosomal oligomers were seen in DNA extracts of testis and isolated spermatocysts (clonal germ cell/ Sertoli cell units) at premeiotic (PrM), but not meiotic (M) or postmeiotic (PoM), stages. Cell nuclei isolated from M stages of development were susceptible to cleavage by micrococcal nuclease, suggesting that developmental control of factors other than a nuclease-insensitive chromatin structure may account for stage specificity. Cytological features of apoptosis were seen in germ cells, but not Sertoli cells, of a subset of isolated PrM spermatocysts and appeared to be all-or-none in affected clones. In culture, DNA fragmentation occurred on schedule with or without various additives, but the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) decreased accumulation of DNA breakdown products. Identification of the apoptotic form of PCD as a major, variable component of normal spermatogenesis and the use of PrM spermatocysts as an in vitro test system will allow further definition of mechanisms and developmental and physiological controls. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 16 (1995) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 184
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    Keywords: IME1 ; Meiosis ; Transcriptional regulation ; S. cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The IME1 gene of Saccharomyces cerevisiae encodes a transcription factor that is required for the expression of meiosis-specific genes. Like many of the genes it regulates, IME1 itself is expressed according to the following complex pattern: barely detectable levels during vegetative growth, and high induced levels under starvation conditions, followed by a subsequent decline in the course of meiosis. This report examines the influence of Ime1 protein on its own expression, demonstrating feedback regulation. Disruption of either IME1 or IME2 leads to constantly increasing levels of Ime1-lacZ expression, under meiotic conditions. This apparent negative regulation is due to cis elements in the IME1 upstream region, which confer transient meiotic expression to heterologous promoter-less genes. A specific DNA/protein complex, whose level is transiently increased under meiotic conditions, is detected on this element. In ime1- diploids, the level of this DNA/protein complex increases, without any decline. These results indicate that the transient expression of IME1 is apparently due to transcriptional regulation. This report also presents evidence suggesting that Ime 1p is directly responsible for regulating its own transcription. Positive feedback regulation in mitotic conditions is suggested by the observation that overexpression of Ime 1p leads to increased levels of IME1-lacZ. Negative autoregulation in meiotic cultures is demonstrated by the observation that a specific point mutation in IME1, ime 1-3, permits expression of meiosis-specific genes, as well as induction of meiosis, but is defective in negative-feedback regulation of IME1. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 16 (1995), S. 264-272 
    ISSN: 0192-253X
    Keywords: Equatorial-less ; compound eye ; genetic mosaics ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: eql (equatorial-less) is a recessive lethal mutation on the second chromosome of Drosophila melanogasfer. J. Campos-Ortega found that eql clones in somatic mosaic flies have reduced numbers of photoreceptor cells, and he suggested that only the R1, R6, and R7 photoreceptor cells were missing in this mutant. These photoreceptor cells help to define the inverted orientation of ommatidial facets along the equatorial midline of the fly eye, hence the mutation was named “equatorial-less”. We have conducted a detailed analysis of the eql mutation, by serial section reconstruction of eql clones marked with bw or w- in somatic mosaic flies. We found that all photoreceptor cell types (Rl-R8) could be deleted by the eql mutation, and in rare cases the number of photoreceptor cells was increased. The apparent lack of photoreceptor cell type specificity was confirmed by our analysis of genetically mosaic facets, which indicated that no single photoreceptor cell, or subset of photoreceptor cells, was uniquely required to express eql Rather, eql appears to function in all photoreceptor cells, and possibly in all eye precursor cells. The distribution of photoreceptor cell numbers in w eql facets was consistent with the hypothesis that each photoreceptor cell was deleted independently of the others. The eql gene is located on the right arm of chromosome 2 at map location 2 - 104.5 ± 0.7 and lies between the polytene chromosome bands 59D8 and 60A7. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 16 (1995), S. 278-290 
    ISSN: 0192-253X
    Keywords: Histone variants ; gene expression ; histone ubiquitination ; H2A ; H2B ; H3 ; sea urchin ; embryogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have used quantitative 2-D protein electrophoresis and immunoprecipitation to study the patterns of histone ubiquitination at 10 h and 36 h of embryonic development in Strongylocentrotus purpuratus. Variants csH2A, αH2A, βH2A, γH2A, δHA, H2AF./Z, αH2B, βH2B, and γH2B showed up to sevenfold differences in level of monoubiquitination between variants, and individual variants showed up to sixfold changes during development. At 36 h of embryogenesis, the late variants were less ubiquitinated than the early variants, althoug h the overall level of ubiquitination was appreciably greater than at 10 h. Antiubiquitin antibodies were used to precipitate formaldehyde-fixed chromatin fragments in order to estimate the degree of ubiquitination of the early histone genes. The 5′ regulatory region of the active H3 gene appeared to be at least twice as ubiquitinated as the adjacent upstream spacer. However, the absolute level of ubiquitination of the early histone gene repeat seemed to be independent of transcriptional activity. These results show that variant-specific ubiquitination of histones is a part of the developmental program in sea urchin embryos, but is not clearly correlated with transcriptional activity of the early histone genes, except perhaps in the regulatory regions. © 1995 Wiley-Liss, Inc.
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  • 187
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    Keywords: Somatic embryogenesis ; temperature-sensitive mutant ts11 ; chitinase ; carrot (Daucus carota L.) ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: At the nonpermissive temperature, somatic embryos of the temperature-sensitive (ts) carrot (Daucus carota L.) cell variant ts11 only proceed beyond the globular embryo stage in the presence of medium conditioned by wild-type cells. The causative component in the conditioned medium has been identified as an acidic 32 kD endochitinase. An antiserum raised against the 32 kD chitinase detected this protein in culture medium from ts11 embryo cultures grown at the permissive temperature as well as at the nonpermissive temperature. No difference in biochemical characteristics or in effect on ts11 embryo development could be detected between the 32 kD chitinase purified from wild-type cultures and the chitinase from ts11 cultures grown at the permissive or at the nonpermissive temperature. Compared to the amount present in a ts11 embryo culture at the permissive temperature, a reduction in the amount of 32 kD chitinase was observed during the temperature-sensitive period at the nonpermissive temperature. These results imply that the arrested embryo phenotype of ts11 is not the result of a structural difference in its 32 kD chitinase, but is the result of a transient decrease in the amount of 32 kD chitinase present. Morphological observations indicate that the ts11 phenotype is pleiotropic and also affects the cell wall of nonembryogenic cells. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 17 (1995), S. 6-15 
    ISSN: 0192-253X
    Keywords: Zebrafish ; epiboly ; gastrulation ; radial intercalation ; cell mixing ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Descendants of early blastomeres in the zebrafish come to populate distinctive regions of the fate map. We present a model suggesting that the distribution of cells in the early gastrula (the fate map stage) results from the passive response of cells to reproducible forces that change the overall shape of the blastoderm just prior to gastrulation. We suggest that one of the morphogenetic changes that accompanies epiboly, the upward doming of the yolk cell into the overlying blastoderm, could be responsible for cell mixing. In support of the model, we show that the timing, extent, and directions of cell mixing in the embryo accurately reflect the expectations of the model. Finally, we show that one portion of the gastrula, a marginal region that later gives rise to many of the mesendodermal derivatives, experiences little cell mixing during the doming process. As a result, this region in the gastrula is populated by the descendants of the subset of the early blastomeres that were originally at the margin. The finding that cytoplasm initially at the edge of the 1-celled blastodisc is transmitted specifically to mesendodermal precursors at the fate map stage raises the possibility that maternal determinants may contribute to initiation of embryonic patterning in the zebrafish embryo. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 17 (1995), S. 55-64 
    ISSN: 0192-253X
    Keywords: Translational control ; activin ; Xenopus ; mesoderm induction ; embryo ; TGF-ß ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Activin is a potent mesoderm inducing factor present in embryos of Xenopus laevis. Recent evidence has implicated activin in the inhibition of neural development in addition to the well-established induction of mesoderm in ectodermal explants. These diverse effects are critically dependent on the concentration of activin yet little is known about the mechanisms regulating the level of activin in the embryo. We report that the 3′ untranslated region (3′ UTR) of activin βB mRNA inhibits the translation of activin in embryos. Microinjection of activin mRNA from which the 3′ UTR has been deleted is 8-10-fold more potent in inducing mesoderm than mRNA containing the 3′ UTR. Truncation of the 3′ UTR also leads to a marked enhancement of activin protein levels in embryos but has no effect when the truncated mRNA is translated in vitro. The 3′ UTR also confers translational inhibition on a heterologous mRNA. These data show that a maternal factor(s) present in X. laevis regulates the translation of injected activin βB mRNA. This factor(s) could be responsible for regulating the levels of endogenous activin βB protein during mesoderm induction and the specification of ectodermal derivatives such as neural and epidermal tissues. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 17 (1995), S. 103-106 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 191
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    Developmental Genetics 17 (1995), S. 129-140 
    ISSN: 0192-253X
    Keywords: Zebrafish ; Danio rerio ; wnt ; pax ; embryogenesis ; neurulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The secreted signalling molecule encoded by the wntl gene and the paired box-containing pax2 gene are thought to play an integral role in patterning the zebrafish rostral nervous system. Using a double-label analysis, we compare the expression patterns of wnt1 RNA and pax2 protein during zebrafish embryogenesis to determine whether they were expressed in identical or overlapping patterns in individual embryos. During gastrulation, wntl RNA was detected in a pattern similar but not identical to the pax2 protein. Later, wntl and pax2 co-localize to the midbrain-hindbrain boundary. Exogenous retinoic acid, a teratogen that is known to affect the formation of the midbrain-hindbrain boundary, has a profound affect on both wntl and pax2 expression at gastrulation. Furthermore, when pax2 is overexpressed in zebrafish embryos, the wntl pattern of expression expands ventrally in the prospective rostral neuroepithelium. Despite the widespread and random distribution of exogenous pax2 RNA, it alone is unable to induce wntl expression in other ec-topic sites. These results are consistent with the coordinate expression of wntl and pax2 being in a pathway responsible for establishing the midbrain-hindbrain boundary and support the earlier interpretation that pax2 may regulate wntl expression [Krauss et al., 1992], although only in a subset of embryonic cells. These data suggest that a predisposition for the regionalization of the central nervous system exists at gastrulation. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 17 (1995), S. 185-187 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
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  • 193
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    Developmental Genetics 17 (1995), S. 206-211 
    ISSN: 0192-253X
    Keywords: Genetic conflict ; parent-offspring conflict ; X-chromosome inactivation ; parental imprinting ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The existence of parentally imprinted gene expression in the somatic tissues of mammals and plants can be explained by a theory of intragenomic genetic conflict, which is a logical extension of classical parent-offspring conflict theory. This theory unites conceptually the phenomena of autosomal imprinting and X-chromosome inactivation. We argue that recent experimental studies of X-chromosome inactivation and andro-genetic development address previously published predictions of the conflict theory, and we discuss possible explanations for the occurrence of random X-inactivation in the somatic tissues of eutherians. © 1995 Wiley-Liss, Inc.
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    Developmental Genetics 17 (1995), S. 263-271 
    ISSN: 0192-253X
    Keywords: Mouse embryos ; SGP-1 mRNA ; antisense ; gene transcription ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a search for genes expressed in preimplantation mouse embryos that are important for the earliest steps in differentiation, we identified an abundant mRNA that codes for a sulfated glycoprotein, SGP-1. The amount of this RNA rises ˜ 100-fold during preimplantation development to a level approximately equal to that of β-actin mRNA in blastocysts, although the level of these transcripts per cell remains fairly constant during these stages at ˜ 2,000-4,000 copies. An antisense RNA that is complementary to approximately the last one-third of the message and contains an open reading frame of 455 nt was found in blastocysts at a 2-3-fold higher level than the mRNA. In situ hybridization with sense and antisense riboprobes showed that both strands are distributed throughout the embryo. The abundance of the SGP-1 mRNA indicates that the encoded protein may play an important role in the development of embryos, and the excess of antisense RNA raises the possibility of an unusual mechanism of regulating its expression. © 1995 Wiley-Liss, Inc.
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  • 195
    ISSN: 0192-253X
    Keywords: Retinoic acid receptors ; retinoic acid ; Xenopus ; CNS ; pattern formation ; ultraviolet ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the isolation of two retinoic acid receptor isoforms (RARγ), which differ only in the 5′ untranslated and putative N-terminus A regions. The two isoforms appear to serve as early markers for the presumptive neural axis; however, their expression patterns differ. RARγ2, 1 is first expressed at gastrulation at the dorsal lip and subsequently along the presumptive neural axis. RARγ2.2 represents the full-length sequence of a receptor cDNA already partially characterized and present as a maternal transcript [Ellinger-Ziegelbauer and Dreyer (1991); Genes Dev 5:94-104, (1993): Mech Dev 41:31-46; Pfeffer and DeRobertis, (1994) Mech Dev: 45:147-153]. Unlike RARγ2.2, the 2.1 variant is not expressed either in pre-somitic mesoderm or notochord. RARγ2.1 is strongly expressed in branchial arches and to a lesser extent in the neural floor plate. The two isoforms also exhibit differential sensitivity to retinoic acid. Constitutive expression of RARγ2.2 following neurulation appears to be depressed by treatment with retinoic acid, but domains of highest expression, namely, the head and tail, remain relatively unaffected, as do patterns of expression prior to late neurulation. By contrast, RARγ2.1 is not transcribed in retinoid-inhibited structures. Using microinjection techniques, we show that changes of RARγ2.1 expression in presumptive head structures occur as an early and local consequence of retinoic acid administration. Since RARγ2.1 expression is inhibited by retinoic acid, we tested to see if other treatments that perturb axis formation had any effect. Surprisingly, UV irradiation did not suppress expression of the RARγ2.1 transcript, suggesting that its inhibition by retinoic acid is not due solely to inhibition of anterior neural development. These experiments demonstrate a new subdivision of isoforms that undergo differential expression during development and that exhibit differential sensitivity to retinoic acid and to UV. This sensitivity and the presence of this isoform variant in regions that are known to exhibit polarizing activity strengthen the hypothesis that these receptors play a primary role during morphogenesis.
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  • 196
    ISSN: 0192-253X
    Keywords: Xenopus laevis ; heat shock protein ; HSP 30 ; developmental gene expression ; translation ; immunoblotting ; microinjection ; embryos ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the present study we have characterized the synthesis of members of the HSP30 family during Xenopus laevis development using a polyclonal antipeptide antibody derived from the carboxyl end of HSP30C. Two-dimensional PAGE/immunoblot analysis was unable to detect any heat-inducible small HSPs in cleavage, blastula, gastrula, or neurula stage embryos. However, heat-inducible accumulation of a single protein was first detectable in early tailbud embryos with an additional 5 HSPs at the late tailbud stage and a total of 13 small HSPs at the early tadpole stage. In the Xenopus A6 kidney epithelial cell line, a total of eight heat-inducible small HSPs were detected by this antibody. Comparison of the pattern of protein synthesis in embryos and somatic cells revealed a number of common and unique heat inducible proteins in Xenopus embryos and cultured kidney epithelial cells. To specifically identify the protein product of the HSP30C gene, we made a chimeric gene construct with the Xenopus HSP30C coding sequence under the control of a constitutive promoter. This construct was microinjected into fertilized eggs and resulted in the premature and constitutive synthesis of the HSP30C protein in gastrula stage embryos. Through a series of mixing experiments, we were able to specifically identify the protein encoded by the HSP30C gene in embryos and somatic cells and to conclude that HSP30C synthesis was first heat-inducible at the early tailbud stage of development. The differential pattern of heat-inducible accumulation of members of the HSP30 family during Xenopus development suggests that these proteins may have distinct functions at specific embryonic stages during a stress response.
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  • 197
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989), S. 542-551 
    ISSN: 0192-253X
    Keywords: Glycine max ; Transposable element ; Transposon tagging ; Genetic instability ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An unstable mutation for anthocyanin pigmentation in soybean (Giycine max [L.] Merr.) was identified in 1983. The mutability is conditioned by an allele at the w4 locus that is recessive to wild type. The population containing the mutable allele is known as the w4-mutable line. Most plants in the line have chimeric flowers with purple sectors on a near-white background. The mutable allele yields germinal revertants at a rate that varies from 5 to 10% per generation, and the revertant alleles are stable. Approximately 1% of the progenies derived from germinal revertant plants contain mutations at other loci These features, as well as the occurrence of pale flower phenotypes and changes of state, suggest that a transposable element system is producing the unstable phenotype.Several new mutants were isolated in an experiment designed to tag loci. The first three chlorophyll-deficient mutants found (CD-1, CD-2, and CD-3) are inherited as single-gene recessives. Each of the mutants lacks the same two mitochondrial malate dehydrogenase (MDH) bands. No recombination has been detected between the MDH phenotype and the chlorophyll-deficient phenotype. Genetic data indicate that the three mutants are allelic, and additional evidence suggests that each of the CD mutants is the result of a deletion. In the CD-1, CD-2, and CD-3 mutants, the deletions result in the silencing of an MDH locus, atypical chloroplast development, and an altered chlorophyll composition. Additional mutants for root necrosis, partial and near sterility, chlorophyll deficiency, and flower color isolated from the transposon tagging study have provided material for future research.
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  • 198
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 2-14 
    ISSN: 0192-253X
    Keywords: Gene expression ; mRNA localization ; ascidian embryos ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have cloned and characterized the temporal and spatial expression of ScCAl5, a cDNA clone encoding an actin gene in the ascidian Styela clava. The partial nucleotide and derived amino acid sequences of this single- copy gene suggest that it is a cytoskeletal actin. Northern analysis shows that ScCAl5 corresponds to a 1.8-kb mRNA that is transcribed during oogenesis, during embryonic development, and in the adult. In situ hybridization shows that maternal ScCA15 mRNA is distributed uniformly in the cyto- plasm of the oocyte and unfertilized egg. During the period of ooplasmic segregation following fertilization, however, ScCAl5 mRNA appears to be translocated into the ectoplasm, a specialized cytoplasmic region of the egg. During the early cleavages, the ectoplasmic transcripts are partitioned to ectodermal cells in the animal hemisphere, which are precursors of the epidermis and nervous system of the larva. Maternal ScCA15 mRNA is degraded just before gastrulation and replaced by zygotic transcripts which begin to accumulate between the neurula and mid-tailbud stages. Zygotic ScCAl5 mRNA accumulates primarily in the epidermal and neural cells, although lower levels of these transcripts may also be present in tail muscle cells. These results show that two mechanisms are used to concentrate ScCA15mRNA in the ectodermal cells during development: (1) localization and differential segregation of maternal transcripts and (2) specific expression of the ScCA15 gene. ScCAl5 mRNA is detected by in situ hybridization in the testes, ovaries, alimentary tract, and endostyle of adults. In the testes, ScCA15 mRNA is present in developing sperm, whereas in the ovary, these transcripts are present in the germinal epithelium and developing oocytes. In the alimentary tract, ScCAl5 mRNA is confined to the gastric epithelium of the esophagus, stomach, and intestine. Since the ScCA15 gene is expressed in embryonic and adult tissues that are undergoing rapid cell division, this actin is likely to function in some aspect of cell proliferation.
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  • 199
    ISSN: 0192-253X
    Keywords: Embryogenesis ; cavitation ; Na ; K-ATPase ; mRNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: NaK-ATPase is a plasma membrane enzyme that plays a critical role in eutherian blastocoel formation (cavitation) by pumping Na+ into the extracellular space enclosed by the trophectoderm. Previous experiments with the mouse had shown that the α (catalytic) subunit of the enzyme becomes detectable by immunocyto-chemistry in the late morula, just prior to the onset of cavitation. In the present study we have used cDNAs corresponding to three mRNA isoforms of the α subunit and a β subunit to determine which genes are expressed during preimplantation development and to explore the timing of their expression. Of the three α subunit cDNAs tested by Northern blot hybridization with blastocyst RNA, only α1 produced a hybridization signal, recognizing a single mRNA about 4 kb in length. This mRNA is relatively abundant in zygotes but barely detectable by the 2-cell stage and then accumulates steadily thereafter to reach its preimplantation maximum in blastocysts. The β1 cDNA detected mRNA of about 2.6-2.8 kb. This mRNA is present in zygotes but could not be detected in 2-, 4-, or 8- cell stages; it is present at a low level in late morulae and is abundant in blastocysts. The temporal profile of accumulation of β1 mRNA thus matches more closely than does α1 the timing of appearance of the catalytic subunit. This suggests that the β subunit may regulate production of the holoenzyme and hence the timing of cavitation.
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  • 200
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 77-87 
    ISSN: 0192-253X
    Keywords: Divergent homeobox ; embryonic transcription ; homeobox evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe two homeobox sequences, TgHbox5 and TgHboxó, isolated from the Hawaiian sea urchin Tripneustes gratilla using a Drosophila Sex combs reduced probe. Sequence analysis shows that the encoded TgHbox5 home-odomain shares only 30-52% amino acid identity with homeodomains encoded by previously characterized genes, establishing that it is a divergent homeobox that is not in any known class of homeoboxes. TgHbox5 is expressed in the embryo as two major developmentally regulated transcripts. one at 5.0 kilobase (kb) appearing by blastula stage and the other at 2.7 kb appearing at pluteus stage. Multiple transcripts from TgHbox5 are present at a much lower level in adult tissues and are predominantly expressed in small and large intestines. The TgHbox6 homeobox is an Antennapedia-class homeobox, which appears not to be expressed during embryogenesis but produces abundant 3.6 and 3.2 kb transcripts in the six adult tissues examined.
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