Localization of mRNA in Tissues of The Chick Embryo By FISH
Mindy George-Weinstein, Ph.D., Department of Anatomy, Philadelphia College of Osteopathic Medicine

Click here for close-up of Images A, B     Click here for close-up of Images C, D     Click here for close-up of Images E, F     Click here for close-up of Images G, H	The stage 14 chick embryo was embedded on paraffin and sectioned transversely at 10µm. In Situ hybridizations were performed (see protocol) using dendrimers containing the fluorochrome Cy3 and an anti-sense oligonucleotide sequence to the housekeeping enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (B), the skeletal muscle specific transcription factor MyoD (D), or embryonic fast myosin (F). Dendrimers lacking a specific recognition sequence (NS) were used as a negative control (H). Differential interference contract images of the sections are shown in A, C, E, and G. The fluorescence photomicrographs are merged images of bisbenzamide labeled nuclei in blue and Cy3 labeled dendrimers in red. Tissues were abundantly labeled with dendrimers to GAPDH. Consistent with previous in situ hybridization results using conventional single stranded oligonucleotide probes, MyoD dendrimers were most heavily concentrated in the dorsal-medial portion of the dermomyotome (dm) of the somite. A few cells in the sclerotome (sc) and neural tube (nt) contained relatively low levels of MyoD mRNA. As expected, the distribution of myosin dendrimers was similar to that of MyoD dendrimers but less abundant. Dendrimers lacking a specific recognition sequence did not bind to this section. Conclusion: Fluorescent dendrimers are sensitive and precise probes for mRNA in tissue sections.

For a full report on the research that produced this data, see:
Gerhart J., Baytion M., DeLuca S., Getts R., Lopez C., Niewenhuis R., Nilsen T, Olex S., Weintraub H., George-Weinstein M., DNA dendrimers localize MyoD mRNA in presomitic tissues of the chick embryo. J Cell Biol. 2000 May 15;149(4):825-34.

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