Abnormal development and dye coupling produced by antisense RNA to gap junction protein in mouse preimplantation embryos.
Antisense RNA to the 27/32-kDa rat liver gap junction (GJ) protein was used to explore the role of GJs in preimplantation embryos. When all blastomeres of two- and four-cell embryos were injected with GJ antisense RNA, the percentage of embryos compacted at 60 hr of development was reduced to less than 20%, while 90% of uninjected embryos and 75% of embryos injected with an unrelated RNA were compacted. When most cells of compacted eight-cell embryos were injected with the GJ antisense RNA, 20% of the embryos were decompacted and only 5% had developed to the blastocyst stage at 90 hr, when blastulation had occurred in 90% of the control embryos. When antisense RNA was injected in one blastomere of four-cell embryos, 40% of the embryos presented a large cell that was not included in the compacted embryo at the time of compaction, and an additional 30% of the embryos had two smaller, excluded blastomeres. These excluded cells were identified as the injected cell with a rhodamine-conjugated dextran marker. To assess effects on junctional communication, one blastomere of some embryos was injected with Lucifer yellow, a GJ-penetrating dye, at various times after a blastomere was injected with antisense RNA. The dye was visible in the whole cell mass of control embryos, but it was excluded from a portion of experimental embryos when the delay between the RNA and the Lucifer yellow injections was 1 hr or longer.
Bevilacqua
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A
Department of Human Genetics, School of Medicine, University of Michigan, Ann Arbor 48109.
Loch-Caruso
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R
Erickson
R P
RP
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D004396:Coloring Agents; D017630:Connexins; D003911:Dextrans; D005260:Female; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D008845:Microinjections; D009924:Organ Culture Techniques; D012313:RNA / Q000008:administration & dosage / Q000235:genetics*; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000037:antagonists & inhibitors*; D012235:Rhodamines
D004396:Coloring Agents; D017630:Connexins; D003911:Dextrans; D008565:Membrane Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D012235:Rhodamines; C051311:rhodamine dextran; D012313:RNA