Genes dependent on zebrafish cyclops function identified by AFLP differential gene expression screen.
Zebrafish cyclops (cyc) encodes a Transforming Growth Factor beta (TGFbeta) signaling factor closely related to mouse Nodal. By comparing amplified fragment length polymorphisms (AFLP) from cyc mutant and wild-type cDNA pools, we devised a differential gene expression screen to isolate genes whose expression is dependent on Cyc signaling. We report two genes not previously described in the zebrafish that were identified using this approach. The first gene, crestin, is expressed predominantly in premigratory and migrating neural crest cells during somitogenesis stages. crestin expression is reduced in cyc mutants initially but recovers by late somitogenesis. The second gene encodes the zebrafish homologue of the calcium-binding protein, calreticulin. Zebrafish calreticulin is highly expressed in the hatching gland and in the floor plate, tissues that are affected in cyc mutants. During gastrulation, calreticulin transcripts are found in the dorsal mesendoderm, in the same cells that express the cyc gene. Expression is reduced in cyc mutants and is abolished by the one-eyed pinhead (oep) mutation that is presumed to prevent Nodal signaling. The identification of calreticulin suggests that a differential screen between wild-type and mutant cDNA is a useful approach to reveal regulation of unexpected gene expression in response to cellular signals. genesis 26:86-97, 2000.
Rubinstein
A L
AL
Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210, USA.
Lee
D
D
Luo
R
R
Henion
P D
PD
Halpern
M E
ME
D000818:Animals; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000502:physiology; D037282:Calreticulin; D018076:DNA, Complementary / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005775:Gastrula / Q000378:metabolism / Q000648:ultrastructure; D005784:Gene Amplification; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005796:Genes; D018398:Homeodomain Proteins / Q000502:physiology; D017403:In Situ Hybridization; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000235:genetics; D005810:Multigene Family; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000502:physiology; D009432:Neural Crest / Q000378:metabolism; D055457:Nodal Protein; D011110:Polymorphism, Genetic; D012261:Ribonucleoproteins / Q000235:genetics* / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics; D014157:Transcription Factors / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000502:physiology*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*; D029961:Zebrafish Proteins
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D018076:DNA, Complementary; D018398:Homeodomain Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D055457:Nodal Protein; C526008:Nodal protein, mouse; D012261:Ribonucleoproteins; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D029961:Zebrafish Proteins; C407354:crestin protein, zebrafish; C512832:ndr2 protein, zebrafish; C110484:tdgf1 protein, zebrafish