Nestin-GFP reporter expression defines the quiescent state of skeletal muscle satellite cells

Abstract

Repair of adult skeletal muscle depends on satellite cells, quiescent myogenic stem cells located beneath the myofiber basal lamina. Satellite cell numbers and performance decline with age and disease, yet the intrinsic molecular changes accompanying these conditions are unknown. We identified expression of GFP driven by regulatory elements of the nestin (NES) gene within mouse satellite cells, which permitted characterization of these cells in their niche. Sorted NES-GFP+ cells exclusively acquired a myogenic fate, even when supplemented with media supporting non-myogenic development. Mutual and unique gene expression by NES-GFP+ cells from hindlimb and diaphragm muscles demonstrated intra- and inter-muscular heterogeneity of satellite cells. NES-GFP expression declined following satellite cell activation and was reacquired in late stage myogenic cultures by non-proliferating Pax7+ progeny. The dynamics of this expression pattern reflect the cycle of satellite cell self-renewal. The NES-GFP model reveals unique transcriptional activity within quiescent satellite cells and permits novel insight into the heterogeneity of their molecular signatures.

Adrenal cortical and chromaffin stem cells: Is there a common progeny related to stress adaptation?

“…All Nestin-GFP cells isolated from either the adrenal medulla or cortex were included in spheres when cultured under low-attachment conditions, which is equivalent to what has been observed with Nestin-GFP cells from this mouse model in other tissues (Birbrair et al, 2011;Day et al, 2007;El-Helou et al, 2013;Li et al, 2003;Mignone et al, 2004;Mignone et al, 2007).…”

“…The intermediate filament Nestin was initially identified as a marker of neural stem and progenitor cells (Cattaneo and McKay, 1990;Lendahl et al, 1990), but has later been shown also to be expressed in stem/progenitor populations in different adult tissues like for example hair follicles (Li et al, 2003;Sellheyer and Krahl, 2010), skeletal muscle (Birbrair et al, 2011;Day et al, 2007) or pancreatic islets (Zulewski et al, 2001). Therefore, it has been suggested that Nestin expression is associated with a stem/progenitor cell population with multipotent properties and regenerative potential (Wiese et al, 2004).…”

Structural-functional organisation of the bone marrow hematopoietic stem cells niches

“…The protein of intermediate filaments nestin (Nes) is expressed by the neuronal cells [161] as well as it is found in various perivascular and endothelial cells [162]. In the BM of transgenic mice with GFP expression under control of Nes-gene regulatory element, the Nes-GFP + cells are localized exclusively on the perivascular area.…”

The A17 enhancer directs expression of Myf5 to muscle satellite cells but Mrf4 to myonuclei

“…Mrf4, but not Myf5, is also implicated in muscle cell differentiation (Bober et al, 1991;Hinterberger et al, 1991). Mrf4 is expressed in adult skeletal muscle (Hinterberger et al, 1991;Gayraud-Morel et al, 2007) and Myf5 in satellite cells (Beauchamp et al, 2000;Day et al, 2007), resident muscle stem cells. Satellite cells (Mauro, 1961) divide to provide myonuclei to the growing myofibers, before becoming quiescent in mature muscle (Schultz et al, 1978), but remain able to again proliferate and differentiate when needed for myonuclear turnover, hypertrophy, and muscle repair/ regeneration (reviewed in Zammit et al, 2006).…”

“…Neither the mouse Myf5 nor the Mrf4 promoter alone is able to drive nlacZ expression in muscle of adult transgenic mice. In conjunction with A17, however, the Myf5 promoter was targeted to satellite cells, mirroring an aspect of the expression domain of the endogenous gene (Beauchamp et al, 2000;Day et al, 2007). Expression was maintained during satellite cell activation and proliferation, but then down-regulated as the cells underwent myogenic differentiation.…”