Localization of specific mRNAs in Xenopus embryos by whole-mount in situ hybridization.
We have adapted a non-radioactive technique to detect localized mRNAs in whole-mount Xenopus embryos. Synthetic antisense RNA transcribed in the presence of digoxygenin-UTP is used as a probe and is detected via an anti-digoxygenin antibody. We show that localized mRNAs can be detected from late gastrula to tadpole stages and that high as well as low abundance RNAs can be detected. The method was tested on muscle actin and alpha-globin RNAs, whose localization has previously been characterized. In addition, we used the method to determine the distribution of XA-1 RNA, an anterior ectoderm-specific RNA, which we show is expressed in the periphery of the cement gland as well as in the region of the hatching gland. The sequence of an XA-1 cDNA predicts a protein rich in proline and histidine.
Hemmati-Brivanlou
A
A
Department of Molecular and Cellular Biology, University of California, Berkeley 94720.
Frank
D
D
Bolce
M E
ME
Brown
B D
BD
Sive
H L
HL
Harland
R M
RM
D000510:Alpha-Globulins / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D006651:Histocytochemistry / Q000379:methods*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics
D000510:Alpha-Globulins; D012333:RNA, Messenger