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Undergraduate Research Project Management System

Expression of MRF4 During Early Xenopus Development

Status Complete
Seeking Researchers No
Start Date 07/01/2008
End Date 06/30/2009
Funding Source Alaska Heart Institute Fellowship
Funding Amount
Community Partner
Related Course
Last Updated 06/26/2009 09:56PM
Keywords xenopus


  Tim Hinterberger

Student Researchers
  Danielle Kusmider


MRF4, Myf5, MyoD, and myogenin, the myogenic regulatory factors (MRFs), are a family of proteins expressed during skeletal muscle development, growth, and regeneration. Although MRF4 is known to function in the maintenance of the differentiated muscle cell type and is expressed during embryonic myogenesis, its role is incompletely understood. Current findings of MRF4 expression in Dr. Hinterberger's lab using Xenopus laevis, has led us to believe that the gene expression is not restricted to the myogenic cells. To further explore the expression of Xenopus MRF4, I will perform additional in situ hybridization on whole embryos brachial arches, otic vesicles, and head mesoderm. I will utilize different probe sequences coding whole-mount in situ hybridization specimens by comparing them to specimens sectioned prior to in situ hybridization. The findings will open doors to future studies of the transcriptional regulation of the MRF4 gene during differentiation of muscle and possibly other cell types. This will lead to better understanding of the regulatory function of MRFs and how genes are controlled.

Specific Aim #1: Determine tissue localization of specific cells of X. leavis that express MRF4 in the head and neck at early neural through metamorphic stages by sectioning specimens of whole mount in situ hybridization. Our recent findings of situ hybridization of whole mount X. laevis embryos contrdict what previous scientists have found. We plan to test and pin point the exact cells of X. laevis which express the MRF4 gene. Furthermore, we will determine whether our results are consistent when probes that correspond to different regions of the MRF4 gene sequence are used.

Specific Aim #2: Determine cell specificity in X. laevis expressing MRF4 in the head and neck at neural through metamorphic stages by section in situ hybridization. To assure that our probe is reaching and binding selectively to the target mRNA sequence of the specimen, we will perform section in situ hybridization. The intensity of staining will reveal whether any of the whole-mount staining is an artifact.

Project goals: The goal of this research project is to further understand the expression of MRF4 during early Xenopus development. This genetic study will present pioneering information about the transcriptional and signaling mechanisms controlling the expression of the MRF4 gene during specification and differentiation of muscle growth. By determining the specific cells expressing the MRF4 gene, we will gain a better understanding of the regulatory functions of the MRF's. This information will allow for further studies on how all genes are controlled and regulated.

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