| dc.description.abstract | A new procedure is described to visualize the spatial pattern of expression of proteins and mRNAs in
cryosections or whole-mounted leech, Drosophila, zebrafish, and chick embryos. Our principal contribution
is in the use of a nonconventional fixation/permeabilization procedure based on the use of formaldehyde
or paraformaldehyde combined with a short C-chain carboxylic acid. Detergents, methanol, and
proteinases were omitted. Hybridization procedures were modified from those of routinely used protocols
developed for the same embryos. Results showed that cytoskeletal and other cytoplasmic proteins, as well
as different mRNAs, were clearly visualized in the expected regions of the embryos. Our procedure has
several advantages over currently used protocols: is simpler, produces better general preservation of
cells, yields reliable results, and can be used for embryos of different taxa at different developmental
stages. It is hypothesized that short C-chain aliphatic carboxylic acids modulate the cross-linking
effect of aldehyde fixatives on cell proteins. Developmental Dynamics 242:503–517, 2013. VC 2013 Wiley
Periodicals, Inc. | |
