KPNA7, an oocyte- and embryo-specific karyopherin α subtype, is required for porcine embryo development
Xin Wang A , Ki-Eun Park A , Stephanie Koser A , Shihong Liu A , Luca Magnani A B and Ryan A. Cabot A CA Department of Animal Sciences, Purdue University, 915 West State Street, West Lafayette, IN 47907, USA.
B Present address: Dartmouth Hitchcock Medical Center, Norris Cotton Cancer Center, One Medical Center Drive, Lebanon, NH 03756, USA.
C Corresponding author. Email: rcabot@purdue.edu
Reproduction, Fertility and Development 24(2) 382-391 https://doi.org/10.1071/RD11119
Submitted: 2 May 2011 Accepted: 15 June 2011 Published: 4 November 2011
Abstract
Coordinated partitioning of intracellular cargoes between nuclear and cytoplasmic compartments is critical for cell survival and differentiation. The karyopherin α/β heterodimer functions to import cytoplasmic proteins that possess classical nuclear localisation signals into the nucleus. Seven karyopherin α subtypes have been identified in mammals. The aim of this study was to determine the relative abundance of transcripts encoding seven karyopherin α subtypes in porcine oocytes and embryos at discrete stages of cleavage development, and to determine the developmental requirements of karypopherin α 7 (KPNA7), an oocyte and cleavage stage embryo-specific karyopherin α subtype. We hypothesised that knockdown of KPNA7 would negatively affect porcine cleavage development. To test this hypothesis, in vitro matured and fertilised porcine oocytes were injected with a double-stranded interfering RNA molecule that targeted KPNA7; nuclei were counted in all embryos 6 days after fertilisation. Embryos injected with KPNA7-interfering RNAs possessed significantly lower cell numbers than their respective control groups (P < 0.05). In vitro binding assays also suggest that KPNA7 may transport intracellular proteins that possess unique nuclear localisation signals. Our data suggest that embryos have differential requirements for individual karyopherin α subtypes and that these karyopherin α subtypes differentially transport intracellular cargo during cleavage development.
Additional keywords: cleavage development, intracellular transport.
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