Cumulative labeling of embryonic mouse neural retina with bromodeoxyuridine supplied by an osmotic minipump.
I describe a method for providing cumulative label of bromodeoxyuridine (BrdU) to mouse embryos. Commercially available osmotic pumps, which release their contents at a steady rate, were implanted subcutaneously in the interscapular space of pregnant mice on embryonic day (E) 9.5-12.5. Survival times varied from 4h to 37 days. Tissues (embryonic and neonatal eyes and maternal intestine) were immunochemically labeled for BrdU and examined histologically. The first detectably labeled cells appeared 4-7h post-implantation (hpi) and all cycling cells were labeled for at least 7 days post-implantation (dpi). Retinal development appeared normal. Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) stained retinas that had been exposed to BrdU showed no more apoptotic cells than those unexposed. I conclude that the maternally implanted osmotic pump successfully provides cumulative BrdU labeling in the mouse embryo.
Neuroscience Program and Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA. mfarah2@jhem.jhmi.edu
Mohamed H
MH
Farah
D000818:Animals; D001973:Bromodeoxyuridine / Q000009:adverse effects / Q000378:metabolism*; D002452:Cell Count / Q000379:methods; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D016923:Cell Death / Q000502:physiology; D016503:Drug Delivery Systems / Q000379:methods*; D004622:Embryo, Mammalian; D005260:Female; D020287:In Situ Nick-End Labeling / Q000379:methods; D007211:Indoles / Q000378:metabolism; D015918:Infusion Pumps, Implantable; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D009474:Neurons / Q000378:metabolism*; D011247:Pregnancy; D012160:Retina / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D013997:Time Factors
D007211:Indoles; C007293:DAPI; D001973:Bromodeoxyuridine