Differential Expression of Utrophin-A and -B Promoters in the Central Nervous System (CNS) of Normal and Dystrophic mdx Mice
Article first published online: 27 MAY 2009
© 2009 The Authors; Journal Compilation © 2009 International Society of Neuropathology
Volume 20, Issue 2, pages 323–342, March 2010
How to Cite
Baby, S. M., Bogdanovich, S., Willmann, G., Basu, U., Lozynska, O. and Khurana, T. S. (2010), Differential Expression of Utrophin-A and -B Promoters in the Central Nervous System (CNS) of Normal and Dystrophic mdx Mice. Brain Pathology, 20: 323–342. doi: 10.1111/j.1750-3639.2009.00275.x
- Issue published online: 2 FEB 2010
- Article first published online: 27 MAY 2009
- Received/accepted 19 March 2009.
- central nervous system;
- mdx mice;
- utrophin-A and -B promoters
Utrophin (Utrn) is the autosomal homolog of dystrophin, the Duchene Muscular Dystrophy (DMD) locus product and of therapeutic interest, as its overexpression can compensate dystrophin's absence. Utrn is transcribed by Utrn-A and -B promoters with mRNAs differing at their 5′ ends. However, previous central nervous system (CNS) studies used C-terminal antibodies recognizing both isoforms. As this distinction may impact upregulation strategies, we generated Utrn-A and -B promoter-specific antibodies, Taqman Polymerase chain reaction (PCR)-based absolute copy number assays, and luciferase-reporter constructs to study CNS of normal and dystrophic mdx mice. Differential expression of Utrn-A and -B was noted in microdissected and capillary-enriched fractions. At the protein level, Utrn-B was predominantly expressed in vasculature and ependymal lining, whereas Utrn-A was expressed in neurons, astrocytes, choroid plexus and pia mater. mRNA quantification demonstrated matching patterns of differential expression; however, transcription–translation mismatch was noted for Utrn-B in caudal brain regions. Utrn-A and Utrn-B proteins were significantly upregulated in olfactory bulb and cerebellum of mdx brain. Differential promoter activity, mRNA and protein expressions were studied in cultured C2C12, bEnd3, neurons and astrocytes. Promoter activity ranking for Utrn-A and -B was neurons > astrocytes > C2C12 > bEnd3 and bEnd3 > astrocytes > neurons > C2C12, respectively. Our results identify promoter usage patterns for therapeutic targeting and define promoter-specific differential distribution of Utrn isoforms in normal and dystrophic CNS.