Abstract
A subtractive-hybridization technique, combined with differential screenings and subsequent whole mount in situ hybridization (ISH) reactions, was used to isolate novel cDNA clones representing developmentally-regulated genes of carp. Small-scale differential screenings of an oocyte and a segmentation-stage cDNA library using oocyte-specific and segmentation stage-specific enriched probes, yielded 75 positive clones. ISH screening showed that 65% (15) of the oocyte-stage clones and 50% (26) of the segmentation-stage clones were indeed stage-specific. Partial sequence analysis suggests that approximately 65% of the 41 stage-specific clones represent novel genes. In addition, an Otxl clone was isolated. Two novel clones and the Otxl clone are of special interest for developmental studies. The clones represent genes that are locally expressed during embryonic development. The expression patterns of Otxl and one of the novel clones suggest functions in specification of the anterior-posterior axis. The three clones provide molecular markers for the study of gastrulation and the patterning of the a-p axis in teleosts.
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Almendral JM, Sommer D, MacDonald-Bravo H, Burckhardt J, Perera J, Bravo R (1988) Complexity of the early genetic response to growth factors in mouse fibroblasts. Mol Cell Biol 8: 2140–2148
Beddington RSP, Smith JC (1993) Control of vertebrate gastrulation: inducing signals and responding genes. Curr Opin Gen Dev 3:655–661
Chang Y, Huang F, Lo T (1994) The complete nucleotide sequence and gene organization of carp (Cyprinus carpio) mitochondrial genome. J Mol Evol 38:138–155
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Coe IR, Munro R, Sherwood NM (1992) Isolation of different brain-specific isotypes of alpha tubulins from chum salmon (Oncorhynchus keta). DNA Seq 3:257–262
Dawid IB (1994) Intercellular signaling and gene regulation during early embryogenesis of Xenopus laevis. J Biol Chem 269: 6259–6262
Druger RK, Fuchs C, Levine EM, Park CY, Matthews JP, Schechter N (1994) Keratins in the goldfish optic nerve: structural complexity and differential expression. J Comp Neurol 340: 269–280
Fisher EM, Beer-Romero P, Brown LG, Ridley A, McNeil JA, Lawrence JB, Willard HF, Bieber FR, Page DC (1990) Homologous ribosomal protein genes on the human X and Y chromosomes: escape from X inactivation and possible implications for Turner syndrome. Cell 63:1205–1218
Fouquet B, Herrmann H, Franz JK, Franke WW (1988) Expression of intermediate filament proteins during development of Xenopus laevis. Development 104:533–548
Giordano S, Glasgow E, Tesser P, Schechter N (1989) A type II keratin is expressed in glial cells of the goldfish visual pathway. Neuron 2:1507–1516
Helde KA, Grunwald DJ (1993) The DVR-1 (Vg1) transcript of zebrafish is maternally supplied and distributed throughout the embryo. Dev Biol 159:418–426
Joly J, Maury M, Joly C, Duprey P, Boulekbache H, Condamine H (1992) Expression of a zebrafish caudal homeobox gene correlates with the establishment of posterior cell lineages at gastrulation. Differentiation 50:75–87
Kessler DS, Melton DA (1994) Vertebrate embryonic induction: mesodermal patterning and neural patterning. Science 266: 596–604
Lemaire P, Garrett N, Kato K, Gurdon JB (1993) Construction of subtracted cDNA libraries enriched for cDNAs for genes expressed in the mesoderm of early Xenopus gastrulae. C R Acad Sci Paris Sci Vie 316:938–944
Li Y, Allende ML, Finkelstein R, Weinberg ES (1994) Expression of two zebrafish orthodenticle-related genes in the embryonic brain. Mech Dev 48:229–244
Meyer A, Biermann CH, Ortí G (1993) The phylogenetic position of the zebrafish (Danio rerio), a model system in developmental biology: an invitation to the comparative method. Proc R Soc London Ser B 252:231–236
Mori H, Miyazaki Y, Morita T, Nitta H, Mishina M (1994) Different spatio-temporal expressions of three otx homeoprotein transcripts during zebrafish embryogenesis. Mol Brain Res 27: 221–231
Neudecker T (1976) Die Embryonalentwicklung des Karpfens (Cyprinus carpio L). Arch Fischereiwiss 27:25–35
Nordnes S, Krauss S, Johansen T (1994) cDNA sequence of zebrafish (Brachydanio rerio) translation elongation factor-1α: molecular phylogeny of eukaryotes based on elongation factor-1α protein sequences. Biochim Biophys Acta 1219:529–532
Pearson W, Lipman D (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Peng G, Taylor JD, Tchen TT (1992) Increased mitochondrial activities in pigmented (melanized) fish cells and nucleotide sequence of mitochondrial large rRNA. Biochem Biophys Res Comm 189:445–449
Powell B, Crocker L, Rogers G (1992) Hair follicle differentiation: expression, structure and evolutionary conservation of the hair type II keratin intermediate filament gene family. Development 114:417–433
Rothstein JL, Johnson D, DeLoia JA, Skowronski J, Solter D, Knowles B (1992) Gene expression during preimplantation mouse development. Genes Dev 6:1190–1201
Sargent TD (1987) Isolation of differentially expressed genes. Methods Enzymol 152:423–432
Sargent TD, Dawid IB (1983) Differential gene expression in the gastrula of Xenopus laevis. Science 222:135–139
Sasai Y, Lu B, Steinbeisser H, Geissert D, Gout LK, De Robertis EM (1994) Xenopus chordin: a novel dorsalizing factor activated by organizer-specific homeobox genes. Cell 79:779–790
Stroband HWJ, Stevens CJM, Kronnie G te, Samallo J, Schipper H, Kramer B, Timmermans LPM (1995) Expression of carpcdx, a caudal homolog, in embryos of the carp, Cyprinus carpio. Roux's Arch Dev Biol 204:369–377
Swalla BJ, Makabe KW, Satoh N, Jeffery WR (1993) Novel genes expressed differently in ascidians with alternate modes of development. Development 119:307–318
Thisse C, Thisse B, Schilling TF, Postlethwait JH (1993) Structure of the snail gene and its expression in wild-type spadetail and no tail mutant embryos. Development 119:1203–1215
Villasante A, Wang D, Dobner P, Dolph P, Lewis SA, Cowan NJ (1986) Six mouse alpha-tubulin mRNAs encode five distinct isotypes: testis-specific expression of two sister genes. Mol Cell Biol 6:2409–2419
Wang Z, Brown DD (1991) A gene expression screen. Proc Natl Acad Sci USA 88:11505–11509
Warga RM, Kimmel CB (1990) Cell movements during epiboly and gastrulation in zebrafish. Development 108:569–580
Weeks DL, Melton DA (1987) A maternal mRNA localized to the vegetal hemisphere in Xenopus eggs codes for a growth factor related to TGF-β. Cell 51:861–867
Wells DE, Bruskin AM, O'Brochta DA, Raff RA (1982) Prevalent RNA sequences of mitochondrial origin in sea urchin embryos. Dev Biol 92:557–562
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Stevens, C.J.M., te Kronnie, G., Samallo, J. et al. Isolation of carp cDNA clones, representing developmentally-regulated genes, using a subtractive-hybridization strategy. Roux's Arch Dev Biol 205, 460–467 (1996). https://doi.org/10.1007/BF00377227
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DOI: https://doi.org/10.1007/BF00377227