Dr. Laura Bridgewater, Department of Microbiology & Molecular Biology
The goal of this project was to characterize the molecular pathway by which nBmp2 disrupts Ca2+ handling in skeletal muscle of nBmp2 mutant mice. Previously, we had worked with Dr. Chad Hancock’s lab to measure muscle function in nBmp2 mutant mice, and those results showed a slowing in the rate at which Ca2+ is pumped from the cytoplasm back into the sarcoplamic reticulum at the conclusion of a skeletal muscle contraction. Our rationale for this MEG study was that characterizing the muscle dysfunction phenotype in the nBmp2 mutant mice would ultimately lead to a molecular level understanding of the functions of nBmp2 in the nucleus. The purpose of this project was achieved successfully, as described below.
Achievement of academic objectives
The primary academic objective of this project was to perform experiments that would complement the muscle stimulation experiments we had already performed with Dr. Chad Hancock, in order have a complete ‘story,’ suitable for publication. This objective was achieved. Chris Fox, an undergraduate student, using ideas from the current literature, developed a protocol for isolating sarcoplasmic reticulum (SR) membranes from mouse muscle, and then he assayed the activity of the SERCA Ca2+ pumps embedded in those membranes. This data became Figure 7 in a manuscript that is ready to submit. Nick Wallace, another undergraduate student, worked with Dr. Allen Parcell to complete a fiber typing analysis of skeletal muscle from the mutant mice compared to controls. This data because Figure 8 in the paper. Chris and Nick are both authors on the manuscript, as is Dr. Parcell. The manuscript (which is included at the end of this report) has been reviewed by all BYU authors. I am preparing to send it to Dr. Mario Capecchi at the University of Utah for his review and comments, and then the manuscript will be submitted for publication. A peer reviewed publication in a scientific journal is the best indication of whether a project’s aims were successfully achieved, and we are well on our way to that publication.
Evaluation of the mentoring environment
My lab operated very well over the past year as a successful mentoring environment. We’ve had an ideal distribution of graduate students (one senior Ph.D. student and one new M.S. student), senior undergraduates who have been in my lab for as much as two years, and new undergraduates who have worked closely with the senior undergraduates to learn techniques, lab safety, and experimental design and interpretation. We hold lab meeting on Friday every week, and each student shows his or her results from the past week’s experiments. If a student has had technical problems, we all help in trouble-shooting. Besides providing each student with regular feedback on their work, this meeting also has been invaluable in broadening each student’s understanding of science in general and of the other projects going on in the lab. It has been gratifying to see the students grow and develop, increasing in independence as they become more comfortable performing techniques and can begin looking at the bigger picture of the work we’re doing. In addition to the lab environment, we’ve had a couple of lab parties at my house that have helped build friendship and unity and have also let students get to know my family and learn more about how I’ve managed to successfully balance school, work, family, and church. These have opened doors for later conversations about decisions and priorities in life, which I hope have been beneficial for the students.
Students involved and academic deliverables produced
Students involved
Student | Current position |
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Trina Loos | Graduated and seeking a postdoc in St. Louis |
Fialka Grigorova | Still in my lab |
Ryan Cordner | Just received a job offer while still completing a clinical lab science internship, applying to Ph.D. programs |
Nick Wallace | Still at BYU, applying to medical schools |
Michael Adam | In medical school |
Broc McCune | In a Ph.D. program at Washington University |
Michael Baldwin | Still at BYU, in my lab, applying to dental schools |
Derek Hill | In medical school |
Caitlin Nichols | On a mission |
Julia Ventura | On a mission |
Brandalyn Andreasen | Still in my lab |
Linsdie Martin | Still in my lab |
Jason Cooper | Still in my lab |
Ben Fisher | Still in my lab |
Jacob Barney | Still in my lab |
Jayson Wurtzbacher | Still in my lab |
Aubrey Rogers | Still in my lab |
Wayna Chow | Still in my lab |
Jordan Richardson | Still in my lab |
Publications in preparation, which have been supported in part by this MEG Grant
- Bridgewater, L.C., Mayo, J.L., Evanson, B.G., Schmidt, A.D., Fox, C.L., Wallace, N.B., Loganathan, S.K., Adam, M.M., Nichols, C.A., Barrow, J.R., Parcell, A.C., Capecchi, M.R., and Hancock, C.R. Nuclear bone morphogenetic protein 2 (nBmp2) mutant mice exhibit impaired skeletal muscle relaxation.
- Cordner, R.D., Ventura, J.S., Mayo, J.L., Andreasen, B., Barrow, J.R., Edwards, J.G., Capecchi, M.R., and Bridgewater, L.C. Mice bearing a targeted mutation of nBmp2 display decreased memory capability. In preparation.
- Loos, T.J., Cordner, R.D., and Bridgewater, L.C. Nuclear Bmp4: A novel SCF E3 ubiquitin ligase interacting protein. In preparation.
Poster presentations
- Loos, T.J., and Bridgewater, L.C. “Nuclear Bmp4 (nBmp4) interacts with the SCF E3 ubiquitin ligase.” American Society for Matrix Biology meeting, Charleston, South Carolina (2010).
- Loos, T.J., and Bridgewater, L.C. “Nuclear Bmp4: a novel ROC1 and ROC2 binding partner.” American Society for Biochemistry and Molecular Biology Annual Meeting, Anaheim, CA (2010).
- Cordner, R.D., Ventura, J., Blickenstaff, J., Walther, C., Mayo, J.L., Felin, J.E., Andreasen, B., Wallace, N., Cappechi, M.R., Edwards, J.G., and Bridewater, L.C. “Mice bearing a targeted mutation of nBmp2 display decreased memory capabilities. American Society for Biochemistry and Molecular Biology Annual Meeting, Anaheim, CA (2010).
- Baldwin, M. and Bridgewater, L.C. “Identification of a consensus DNA binding sequence for nGdf5.” American Society for Biochemistry and Molecular Biology Annual Meeting, Anaheim, CA (2010).
- McCune, B.T., Finley, M., Schmidt, A.D., Fox, C., Mayo, J.L., and Bridgewater, L.C. “Binding of nBmp2 to PLSCR1 suggests a possible mechanism for nBmp2 regulation of calcium- modulating proteins.” American Society for Biochemistry and Molecular Biology Annual Meeting, Anaheim, CA (2010).
- Evanson, B.G., Schmidt, A.D., Mayo, J.L., Bridgewater, L.C., and Hancock, C.R.. “Nuclear bone morphogenetic protein 2 mutant mice exhibit slowed skeletal muscle relaxation and decreased tetany in the gastrocnemius, plantaris, soleus muscle complex.” American Society for Biochemistry and Molecular Biology Annual Meeting, Anaheim, CA (2010).
Grant proposal
The results obtained in research supported by this MEG funding provided a large part of the Preliminary studies portion of an NIH grant proposal submitted October 25, 2010, entitled “A novel nuclear variant of nBmp2: role in Ca2+ transport & cell cycle regulation.” If funded on the first submission, this grant would provide $450,000 in total costs over a three year period, beginning in July of 2011.
Description of Results/Findings
The muscle findings that resulted from this work are described in the manuscript mentioned above and attached. As a bonus, this work regarding Ca2+ transport in skeletal muscle led us to wonder whether Ca2+ transport is also disrupted in neural signaling in the nBmp2 mutant mice. Ryan Cordner performed some behavioral studies that showed learning/memory problems in the mutants, and we are now working with Dr. Jeff Edwards of the PDBIO Department to measure long term potentiation (LTP) in hippocampal slices from control and mutant mice. Preliminary results suggest that the mutant mice do have problems with Ca2+ transport in neurons as well as in muscle. We plan to submit this work for publication after the LTP studies are complete.
Summary of Spending
The following students were paid wages from this MEG project funding between January and September of 2010, at which time the funds were exhausted:
- Ryan Cordner, $8.50/hr, up to 20 hrs/week winter semester and up to 40 hrs/week spring term
- Michael Adam, $8.50/hr, up to 10 hrs/week, winter semester
- Michael Baldwin, $8.5/hr, up to 10 hrs/week, winter semester
Other students who worked using supplies paid for by this MEG funding (listed above) were paid wages from another account, which is now also exhausted.
Total spending:
Student wages | $4,113 |
Student benefits | $86 |
Supplies | $15,512 |
Postage/mailing | $71 |
RIC Facility usage fees | $180 |
Equipment maintenance | $38 |
Total | $20,000 |
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