Changing intracellular compartmentalization of beta-galactosidase in the ROSA26 reporter mouse during embryonic development: a light- and electron-microscopic study.
The beta-geo (LacZ) reporter gene encodes for beta-galactosidase (beta-gal) in all cells of the ROSA26 mouse during embryonic development. As such, beta-gal activity constitutes an excellent marker for in situ labeling of expressing cells. However, the intracellular distribution of beta-gal differs between cells, and changes during embryonic development. Therefore, we studied LacZ-encoded beta-gal using light and electron microscopy in the heart, lung, liver, and small intestine on days 13 and 16 of gestation, and the kidney on day 16 of gestation in ROSA26 mice. The Bluo-gal method was carried out under standardized conditions, including fixation, washing, and incubation procedures. Intracellular beta-gal staining is encountered in a combination of membranous compartments, including the nuclear envelope, the endoplasmic reticulum, and the plasma membrane. Its exact localization depends on the cell type and is regulated during development. Therefore, one must take the compartmental transition of intracellular beta-gal staining into consideration when interpreting results obtained from experiments using ROSA26 mice.
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.
Naoyoshi
N
Aoyama
Daniel G M
DG
Molin
Monica M T
MM
Mentink
Henk K
HK
Koerten
Marco C
MC
De Ruiter
Adriana C
AC
Gittenberger-De Groot
Robert E
RE
Poelmann
D000818:Animals; D004622:Embryo, Mammalian / Q000201:enzymology; D047108:Embryonic Development; D017930:Genes, Reporter; D006651:Histocytochemistry; D051379:Mice; D008815:Mice, Inbred Strains / Q000196:embryology*; D008854:Microscopy, Electron; D011506:Proteins / Q000235:genetics*; D022661:RNA, Untranslated; D014018:Tissue Distribution; D001616:beta-Galactosidase / Q000378:metabolism*
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D011506:Proteins; D022661:RNA, Untranslated; D001616:beta-Galactosidase