Our current work focuses primarily on understanding the differences in microtubule organization between muscles of control mice (WT) and of mdx mice, which serve as a model for Duchenne muscular dystrophy (DMD). Normal mouse muscles have a regular grid-like microtubule network, whereas mdx mouse muscles have a disordered, denser network. The software TeDT, developed in the Light Imaging Section for the analysis of microtubule directionality, is an essential tool in the quantitative assessment of such differences in microtubule organization (Liu et al., 2014). Results obtained in previous years demonstrated that the differences in microtubule orientation can be observed as soon as microtubules start growing from the nucleating Golgi elements. Thus muscle microtubules grow as if Golgi elements themselves, or the nucleating molecules anchored to the Golgi elements, had a specific orientation, disturbed in mdx muscles. Conventional and super-resolution microscopy have been used to investigate the orientation of the Golgi elements. Labeling of the Golgi elements with two antibodies, one for the cis-Golgi protein GM130, the other one for the trans-Golgi protein TGN38 allowed us to determine the orientation of each Golgi element. Plotting Golgi directionality with the software TeDT indeed shows differences between WT and mdx mouse muscles, confirming that differences in microtubule organization are sealed at an early stage of their formation. We have also carried out RNA-seq analysis of three different mouse muscles (FDB, EDL, and soleus) at two different ages, 2 and 5 months, from both WT and mdx mice. The goal was a comparison of RNA changes in the mdx mouse muscles compared to human DMD muscles (Khairallah et al. 2012) with special attention to tubulins, the constituents of microtubules, and microtubule-associated proteins. The two ages were selected based on the report in the same paper that microtubules are implicated in the mdx pathology at 5 but not at 2 months of age. The results are still under analysis. They already confirm that several tubulin mRNAs are differentially expressed in mdx compared to WT muscles. In contrast, centrosomal proteins, involved in microtubule nucleation, and MAPs (microtubule-associated proteins) are little affected. Although several mRNAs are differentially expressed in 2 and 5 mo-old mice, this is not the case for tubulin mRNAs. Thus, the difference in response between these ages must be searched in other parts of the pathways affected. After analysis of the results is complete we will further investigate the role of specific molecules hypothesized to play a role in the disorganization of the mdx network.
| Iglesias-Bartolome, Ramiro; Uchiyama, Akihiko; Molinolo, Alfredo A et al. (2018) Transcriptional signature primes human oral mucosa for rapid wound healing. Sci Transl Med 10: |
| Duverger, Olivier; Carlson, Jenna C; Karacz, Chelsea M et al. (2018) Genetic variants in pachyonychia congenita-associated keratins increase susceptibility to tooth decay. PLoS Genet 14:e1007168 |
| Tsai, Pei-Fang; Dell'Orso, Stefania; Rodriguez, Joseph et al. (2018) A Muscle-Specific Enhancer RNA Mediates Cohesin Recruitment and Regulates Transcription In trans. Mol Cell 71:129-141.e8 |
| Lim, Jeong-A; Sun, Baodong; Puertollano, Rosa et al. (2018) Therapeutic Benefit of Autophagy Modulation in Pompe Disease. Mol Ther 26:1783-1796 |
| Madsen, Agnete B; Knudsen, Jonas R; Henriquez-Olguin, Carlos et al. (2018) ?-Actin shows limited mobility and is required only for supraphysiological insulin-stimulated glucose transport in young adult soleus muscle. Am J Physiol Endocrinol Metab 315:E110-E125 |
| Ferdinand, John R; Richard, Arianne C; Meylan, Françoise et al. (2018) Cleavage of TL1A Differentially Regulates Its Effects on Innate and Adaptive Immune Cells. J Immunol 200:1360-1369 |
| Duverger, Olivier; Cross, Michael A; Smith, Frances J D et al. (2018) Enamel anomalies in a pachyonychia congenita patient with a mutation in KRT16. J Invest Dermatol : |
| Milgroom, Andrew; Ralston, Evelyn (2016) Clearing skeletal muscle with CLARITY for light microscopy imaging. Cell Biol Int 40:478-83 |
| Suzuki, Ryo; Leach, Sarah; Liu, Wenhua et al. (2014) Molecular editing of cellular responses by the high-affinity receptor for IgE. Science 343:1021-5 |
| Feeney, Erin J; Austin, Stephanie; Chien, Yin-Hsiu et al. (2014) The value of muscle biopsies in Pompe disease: identifying lipofuscin inclusions in juvenile- and adult-onset patients. Acta Neuropathol Commun 2:2 |
Showing the most recent 10 out of 19 publications
