Progress 09/01/09 to 08/31/13
Outputs
Target Audience: The primary audience reached by our work are peer scientists in the field of reproductive biology. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Graduate student trainee and doctoral candidate, Yanfang Li, presented her work at one scientific conference in July 2013 and published one manuscript in 2013 regarding her work in this project. How have the results been disseminated to communities of interest? Yes, see presentation at scientific conferece (46th annual meeting for the Society for the Study of Reproduction held in July 2013); we have also published one manuscript in 2013 outlining the data we obtained in completeing this project. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Results of our work show that intracellular proteins that bear nuclear localizaiton signals adopt differential patterns of nuclear localization at discrete stages of cleavage development. These findings suggest that nuclear trafficking pathways serve a role to regulate the intracellular trafficking of discrete transcription factors and chromatin modifying enzymes.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2013
Citation:
Li, Y., K-E. Park, R.A. Cabot. Dynamic changes in nuclear import of a NLS-bearing substrate during the 8-cell stage porcine embryo. Reproduction, Fertility and Development, in press.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
Li, Y. and R.A. Cabot. Restricted nuclear import of an NLS-bearing protein in porcine cleavage stage embryos. 46th annual meeting of the Society for the Study of Reproduction (SSR); abstract 704.
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Progress 09/01/11 to 08/31/12
Outputs
OUTPUTS: We have determined how multiple nuclear trafficking receptors function in transporting chromatin remodeling factors and other nuclear proteins to the nucleus. Using an RNAi-mediated knockdown approach, we find that known importin alpha cargoes fail to adopt a nuclear localization in porcine oocytes. We have also found evidence that suggests that an oocyte-specific importin alpha subtype binds unique clusters of intracellular proteins. PARTICIPANTS: Yangfang Li, graduate student Dominique Hall, graduate student Xin Wang, graduate student Kieun Park, postdoctoral scientist Shihong Liu, graduate student TARGET AUDIENCES: Scientific audiences were reached by publishing in scientific journals. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Results of this work suggest that nuclear trafficking receptors regulate access of nuclear factors in porcine oocytes and cleavage stage embryos and thereby serve regulatory role in porcine embryo development.
Publications
- Park, K-E., Inerowicz, H.D., Wang, X., Li, Y., Koser, S., and Cabot, R.A. 2012. Identification of Karyopherin α1 andα7 interacting proteins in porcine tissue. PLoS One 7(6) e38990.
- Wang X., Park, K-E., Koser, S., Liu, S., Magnani, L., and Cabot, R.A. 2012. KPNA7, an oocyte- and embryo-specific karyopherin α subtype, is required for porcine embryo development. Reproduction, Fertility and Development, 24(2):382-391.
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Progress 09/01/10 to 08/31/11
Outputs
OUTPUTS: We have developed our microinjection assay and successfully knocked down individual KPNA subtypes in porcine oocytes and embryos and determined the developmental phenotype for 4 of the 7 KPNA subtypes. We have determined the import dynamics of three cargoes that are trafficked by specific KPNA subtypes. PARTICIPANTS: Postdoctoral, graduate and undergraduate student training has involved the following individuals: Kieun Park, postdoctoral associate; Kallie Foust, graduate student; Dominique Hall, graduate student; Emily Vargo, graduate student; Katelyn Jackson, undergradute student. TARGET AUDIENCES: Reproductive biologists working in the feild of mammalian embryo development. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Our findings have shown us that individual KPNA subtypes, while detectable at the mRNA and protein levels in oocytes and cleavage stage embryos, adopt unique intracelllular localization patterns during cleavage development. This finding suggests that cargoes trafficked by specific KPNA subtypes my be limited in their ability to access chromatin during cleavage development.
Publications
- No publications reported this period
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Progress 09/01/09 to 08/31/10
Outputs
OUTPUTS: To date we have determined that there appears to be an abrupt change in the function of specific nuclear transport systems during progression from the immature oocyte to the 4-cell stage embryo. During this time several key events occur to the oocyte and embyro that have profound effects on the ability of the embryo to continue development. Identifying that a nuclear transport system changes in functional will provide insight into what intracellular proteins can access the nucleus at key points in development. Such knowledge has been communicated at an annual scientific meeting for the Society for the Study of Reproduction and at an invited research presentation in Copenhagen, Denmark at the Univeristy of Copenhagen, for the faculties of Animal Sciences and Veterinary Medicine. PARTICIPANTS: Yangfang Li, graduate student Xin Wang, PhD candidate Ki-Eun Park, postdoctoral scientist TARGET AUDIENCES: Attendees at the annual meeting for the Society for the Study of Reproduction (July 30-August 3, 2010). PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Much of the work we have undertaken in the first year of this project has centered on Specific Aims 1 and 3. We have determined the intracellular localization of three importin alpha subtypes (KPNA1, 2 and 7) in porcine oocytes and cleavage stage embyros, we have also developed specific porcine antibodies that recognize 5 of the 7 porcine importin alpha isoforms; these reagents will be for key importance as we proceed to Specific Aim 2 of this proposal and dissect the developmental requirments of individual importin alpha isoforms. In Specific Aim 3 we have determined that transprot mediated by the importin alpha subtype known as KPNA 1 apprears to change in function during cleavage, being unable to trasport intracellular cargo until the 2-cell stage of development. Current studies are underway to confirm this finding. We are also extending our efforts to evalute transport mediate by other KPNA subtypes. Students and postdoctoral students have have contributed to this porject during this time period include the following: Xin Wang, PhD candidate Yanfang Li, gradute student Ki-Eun Park, postdoctoral researcher
Publications
- Li, Y., K.E. Park, and R.A. Cabot. 2010. Recombinant Sox2 protein rarely adopts a nuclear localization in immature porcine oocytes. Society for the Study of Reproduction (SSR) July 30-August 3, 2010.
- Wang, X., K.E. Park, K. Foust, and R.A. Cabot. 2010. Karyopherin Alpha 1, 2, and 7 Adopt a Nuclear Localization in Porcine Oocytes and Cleavage Stage Embryos. Society for the Study of Reproduction (SSR) July 30-August 3, 2010.
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