title: Automated microinjection of recombinant BCL-X into mouse zygotes enhances embryo development.
abstract: Progression of fertilized mammalian oocytes through cleavage, blastocyst formation and implantation depends on successful implementation of the developmental program, which becomes established during oogenesis. The identification of ooplasmic factors, which are responsible for successful embryo development, is thus crucial in designing possible molecular therapies for infertility intervention. However, systematic evaluation of molecular targets has been hampered by the lack of techniques for efficient delivery of molecules into embryos. We have developed an automated robotic microinjection system for delivering cell impermeable compounds into preimplantation embryos with a high post-injection survival rate. In this paper, we report the performance of the system on microinjection of mouse embryos. Furthermore, using this system we provide the first evidence that recombinant BCL-XL (recBCL-XL) protein is effective in preventing early embryo arrest imposed by suboptimal culture environment. We demonstrate that microinjection of recBCL-XL protein into early-stage embryos repairs mitochondrial bioenergetics, prevents reactive oxygen species (ROS) accumulation, and enhances preimplantation embryo development. This approach may lead to a possible treatment option for patients with repeated in vitro fertilization (IVF) failure due to poor embryo quality.
authors.affiliation: Department of Mechanical and Industrial Engineering and Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Xinyu
authors.initials: X
authors.lastname: Liu
authors.forename: Roxanne
authors.initials: R
authors.lastname: Fernandes
authors.forename: Marina
authors.initials: M
authors.lastname: Gertsenstein
authors.forename: Alagammal
authors.initials: A
authors.lastname: Perumalsamy
authors.forename: Ingrid
authors.initials: I
authors.lastname: Lai
authors.forename: Maggie
authors.initials: M
authors.lastname: Chi
authors.forename: Kelle H
authors.initials: KH
authors.lastname: Moley
authors.forename: Ellen
authors.initials: E
authors.lastname: Greenblatt
authors.forename: Igor
authors.initials: I
authors.lastname: Jurisica
authors.forename: Robert F
authors.initials: RF
authors.lastname: Casper
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.forename: Andrea
authors.initials: A
authors.lastname: Jurisicova
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001331:Automation; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D051379:Mice; D008845:Microinjections / Q000295:instrumentation / Q000379:methods*; D011994:Recombinant Proteins / Q000008:administration & dosage* / Q000494:pharmacology*; D012371:Robotics; D015996:Survival Rate; D015053:Zygote / Q000166:cytology / Q000187:drug effects*; D051020:bcl-X Protein / Q000008:administration & dosage* / Q000494:pharmacology*
chemical_list: D011994:Recombinant Proteins; D051020:bcl-X Protein