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TACC3 expression and localization in the murine egg and ovary
Zhonglin Hao1, Mark H Stoler, Buer Sen
1Department of Cell Biology and Center for Recombinant Gamete Immunocontraceptive Vaccinogens, University of Virginia, Charlottesville, Virginia 22908, USA.
Molecular Reproduction and Development
|September 19, 2002
Summary
Transforming Acidic Coiled Coil containing protein (TACC3) was identified in mouse oocytes. Two spliced forms, TACC3a and TACC3b, were found, with TACC3a showing wide tissue distribution and cytoplasmic localization in growing oocytes.
Area of Science:
- Molecular Biology
- Cell Biology
- Reproductive Biology
Background:
- Transforming Acidic Coiled Coil containing protein (TACC3) is a protein identified in mouse oocytes.
- TACC3 shares homology with other TACC family members and Xenopus Maskin, a factor associated with cytoplasmic polyadenylation element binding protein (CPEB).
Purpose of the Study:
- To identify and characterize the protein TACC3 in immature mouse oocytes.
- To investigate the different spliced forms of TACC3 and their expression patterns.
- To determine the subcellular localization and tissue distribution of TACC3.
Main Methods:
- Protein identification using tandem mass spectrometry on 2D gel-separated proteins from mouse oocytes.
- Polymerase Chain Reaction (PCR) amplification to identify alternatively spliced forms.
- Northern blot analysis for tissue expression profiling.
- Transient transfection of CV-1 cells to determine protein localization.
- In situ hybridization on mouse ovarian tissue sections for cellular localization.
Main Results:
- TACC3 was identified as a protein in germinal vesicle stage mouse oocytes.
- Two alternatively spliced forms, TACC3a (630 aa, 70 kDa) and TACC3b (426 aa, 49 kDa), were identified.
- TACC3a exhibited broad tissue distribution, including high expression in testis and spleen, and moderate expression in ovary, heart, and lung.
- Both TACC3a and TACC3b were localized to the cytoplasm in transfected cells.
- In situ hybridization revealed abundant TACC3 expression in the cytoplasm of growing oocytes within the ovary.
Conclusions:
- TACC3 is present in mouse oocytes and exists in at least two alternatively spliced forms.
- The expression pattern suggests TACC3 plays a role in ovarian cells during active growth and development.
- TACC3's cytoplasmic localization in oocytes indicates potential involvement in cytoplasmic processes during oocyte maturation.