ZFIN ID: ZDB-PUB-011217-10
In situ hybridization screen in zebrafish for the selection of genes encoding secreted proteins
Crosier, P.S., Bardsley, A., Horsfield, J.A., Krassowska, A.K., Lavallie, E.R., Collins-Racie, L.A., Postlethwait, J.H., Yan, Y.-L., McCoy, J.M., and Crosier, K.E.
Date: 2001
Source: Developmental dynamics : an official publication of the American Association of Anatomists   222(4): 637-644 (Journal)
Registered Authors: Bardsley, Anne, Crosier, Kathy, Crosier, Phil, Horsfield, Jules, Postlethwait, John H., Yan, Yi-Lin
Keywords: expression screen, secreted proteins, signal sequence trap, zebrafish, mutant embryos, expression pattern, signal sequence, Brachyury gene, tail formation, C. elegans, mouse, Xenopus, vertebrate, Drosophila
MeSH Terms:
  • Animals
  • Brain/embryology
  • Chromosome Mapping
  • Embryo, Nonmammalian/metabolism
  • Embryo, Nonmammalian/physiology
  • Eye/embryology
  • In Situ Hybridization
  • Nervous System/embryology
  • Notochord/metabolism
  • Protein Sorting Signals
  • Proteins/genetics*
  • Proteins/metabolism*
  • Tail/embryology
  • Zebrafish/embryology*
  • Zebrafish/genetics*
PubMed: 11748832 Full text @ Dev. Dyn.
An in situ hybridization expression screen using a signal sequence trap system has been conducted in zebrafish to isolate cDNAs that encode secreted proteins. Random clones (secreted expressed sequence tags; sESTs) were sequenced from zebrafish embryonic (18-24 hr postfertilization) and adult kidney libraries. From the two RNA sources, 627 random sEST cDNAs were identified as being homologous or identical to known genes and 166 clones encode currently unidentified genes. The sESTs represent a broad range of enzymes and other regulatory molecules. Whole-mount in situ hybridization analysis was carried out by using antisense probes generated from 244 selected sESTs, and a range of expression patterns was obtained. Genetic mapping undertaken with sEST sequences demonstrated that assignment of map position was attainable by using 5 ' primers. The signal sequence trap system used in this work has yielded a range of cDNAs that encode secreted proteins and, together with analysis of patterns of expression and genetic mapping, has the potential to facilitate analysis of signaling pathways central to development and physiology.