Distribution of RNA binding protein MOEP19 in the oocyte cortex and early embryo indicates pre-patterning related to blastomere polarity and trophectoderm specification.
We report the cloning and characterization of MOEP19, a novel 19 kDa RNA binding protein that marks a defined cortical cytoplasmic domain in oocytes and provides evidence of mammalian oocyte polarity and a form of pre-patterning that persists in zygotes and early embryos through the morula stage. MOEP19 contains a eukaryotic type KH-domain, typical of the KH-domain type I superfamily of RNA binding proteins, and both recombinant and native MOEP19 bind polynucleotides. By immunofluorescence, MOEP19 protein was first detected in primary follicles throughout the ooplasm. As oocytes expanded in size during oogenesis, MOEP19 increased in concentration. MOEP19 localized in the ovulated egg and early zygote as a symmetrical spherical cortical domain underlying the oolemma, deep to the zone of cortical granules. MOEP19 remained restricted to a cortical cytoplasmic crescent in blastomeres of 2-, 4- and 8-cell embryos. The MOEP19 domain was absent in regions underlying cell contacts. In morulae, the MOEP19 domain was found at the apex of outer, polarized blastomeres but was undetectable in blastomeres of the inner cell mass. In early blastocysts, MOEP19 localized in both mural and polar trophectoderm and a subset of embryos showed inner cell mass localization. MOEP19 concentration dramatically declined in late blastocysts. When blastomeres of 4- to 8-cell stages were dissociated, the polarized MOEP19 domain assumed a symmetrically spherical localization, while overnight culture of dissociated blastomeres resulted in formation of re-aggregated embryos in which polarity of the MOEP19 domain was re-established at the blastomere apices. MOEP19 showed no evidence of translation in ovulated eggs, indicating that MOEP19 is a maternal effect gene. The persistence during early development of the MOEP19 cortical oocyte domain as a cortical crescent in blastomers suggests an intrinsic pre-patterning in the egg that is related to the apical-basolateral polarity of the embryo. Although the RNAs bound to MOEP19 are presently unknown, we predict that the MOEP19 domain directs RNAs essential for normal embryonic development to specific locations in the oocyte and early embryo.
Herr
John C
JC
Center for Research in Contraceptive and Reproductive Health, Department of Cell Biology, P.O. Box 800732, University of Virginia Health System, Charlottesville, VA 22908, USA. jch7k@virginia.edu
Chertihin
Olga
O
Digilio
Laura
L
Jha
Kula N
KN
Vemuganti
Soumya
S
Flickinger
Charles J
CJ
D000595:Amino Acid Sequence; D000818:Animals; D001757:Blastomeres / Q000166:cytology / Q000502:physiology*; D016764:Cell Polarity; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004475:Ectoderm / Q000166:cytology / Q000502:physiology*; D004527:Egg Proteins / Q000032:analysis; D004622:Embryo, Mammalian / Q000502:physiology; D047108:Embryonic Development; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D013058:Mass Spectrometry; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology / Q000502:physiology*
D004527:Egg Proteins; C527277:MOEP19 protein, mouse; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D012313:RNA; D008715:Methionine