Gene transfer of lacZ into avian neural tube and neural crest cells by retroviral infection of grafted embryonic tissues.
We describe here a new method for transferring genes into cells of the neural tube and neural crest of early avian embryos in vivo. Using the marker gene lacZ as an example, we infected dissected neural tubes from Hamburger-Hamilton stage 12-13 quail embryos with a replication-defective retrovirus carrying lacZ during a 2 hr period of exposure to the virus in culture. Infected neural tubes were then grafted into uninfected host chicken embryos in ovo and, after continued development for several days, the chimeric embryos were processed for beta-galactosidase histochemistry to identify the progeny of infected cells. We show that virus-infected neural tubes grafted isotopically into the trunk region of host embryos gave rise to cells of both the spinal cord and neural crest. Infected neural crest cells localized within dorsal root ganglia, sympathetic ganglia, peripheral nerves, and within the skin, where they were likely to give rise to melanocytes. These data are consistent with those using other cell marking techniques applied to the neural crest, indicating that retrovirus infection in culture, grafting, and beta-galactosidase expression has a neutral effect on neural crest cell migration and localization. These results indicate the heterospecific grafting of early avian tissues infected with retroviruses carrying foreign genes may be an effective strategy for testing the biological role of various gene products during development.
Department of Cell Biology & Anatomy, Oregon Health Sciences University, Portland 97201-3098.
K M
KM
Stocker
A M
AM
Brown
G
G
Ciment
D000818:Animals; D002490:Central Nervous System / Q000166:cytology / Q000196:embryology*; D002642:Chick Embryo; D003370:Coturnix / Q000196:embryology; D046508:Culture Techniques; D016332:Fetal Tissue Transplantation; D007763:Lac Operon; D009417:Nerve Tissue / Q000637:transplantation; D009432:Neural Crest / Q000166:cytology / Q000502:physiology*; D010063:Ovum / Q000502:physiology; D012192:Retroviridae Infections / Q000235:genetics*; D014162:Transfection / Q000379:methods*