High resolution ultrasound-guided microinjection for interventional studies of early embryonic and placental development in vivo in mice

BMC Developmental Biology
John C SlevinS Lee Adamson

Abstract

In utero microinjection has proven valuable for exploring the developmental consequences of altering gene expression, and for studying cell lineage or migration during the latter half of embryonic mouse development (from embryonic day 9.5 of gestation (E9.5)). In the current study, we use ultrasound guidance to accurately target microinjections in the conceptus at E6.5-E7.5, which is prior to cardiovascular or placental dependence. This method may be useful for determining the developmental effects of targeted genetic or cellular interventions at critical stages of placentation, gastrulation, axis formation, and neural tube closure. In 40 MHz ultrasound images at E6.5, the ectoplacental cone region and proamniotic cavity could be visualized. The ectoplacental cone region was successfully targeted with 13.8 nL of a fluorescent bead suspension with few or no beads off-target in 51% of concepti microinjected at E6.5 (28/55 injected). Seventy eight percent of the embryos survived 2 to 12 days post injection (93/119), 73% (41/56) survived to term of which 68% (38/56) survived and appeared normal one week after birth. At E7.5, the amniotic and exocoelomic cavities, and ectoplacental cone region were discernable. Our success at target...Continue Reading

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Citations

Aug 6, 2010·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Vicki PlaksMichal Neeman
Jan 4, 2008·ILAR Journal·Brenda A Klaunberg, Judith A Davis
Aug 7, 2012·Interface Focus·F Stuart FosterS Lee Adamson
Dec 14, 2011·Journal of Biomedicine & Biotechnology·A GrecoM Salvatore
Jul 28, 2006·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Mary E Dickinson

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Methods Mentioned

BETA
dissection
cesarean section
transfection
dissections

Software Mentioned

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