Distribution of type II collagen mRNA in Xenopus embryos visualized by whole-mount in situ hybridization.
We have developed a whole-mount histochemical method to monitor the distribution of expressed genes within the intact, developing vertebrate embryo. Background problems that result from alkaline phosphatase- or horseradish peroxidase-based stains have been minimized, enabling both early and late stages of Xenopus embryogenesis to be monitored. The feasibility and utility of this non-isotopic method has been demonstrated by using a specific DNA probe to localize Xenopus laevis Type II collagen mRNA expression to areas surrounding the vacuoles of the notochord in Stage 30 embryos. Expression expands by Stage 41/42 to form a visually striking distribution pattern that includes a variety of chondrogenic tissues such as the vertebrae, otocysts, mandible, and periocular region. Although these experiments focused on expression of a structural gene, the high resolution and sensitivity of the method should allow it also to monitor expression of less abundant mRNA products of non-structural genes such as transcription factors, cytoplasmic regulators, and growth factors. In addition, this approach should be a successful tool to probe expression in normal and perturbed embryos not only of amphibians but also of other vertebrates, including avians and mammals.
Bieker
J J
JJ
Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York 10029-6574.
Yazdani-Buicky
M
M
D000818:Animals; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics*; D015342:DNA Probes; D004625:Embryo, Nonmammalian / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D009672:Notochord / Q000737:chemistry / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D014982:Xenopus laevis
D015342:DNA Probes; D012333:RNA, Messenger; D003094:Collagen