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.
Farah
Mohamed H
MH
Neuroscience Program and Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA. mfarah2@jhem.jhmi.edu
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