Summary The induction of pluripotency or trans-differentiation of one cell type to another can be accomplished with cell lineage-specific transcription factors. Here we report that repression of a single RNA binding protein PTB, which occurs during normal brain development via the action of miR-124, is sufficient to induce trans-differentiation of fibroblasts into functional neurons. Besides its traditional role in regulated splicing, we show that PTB has a previously undocumented function in the regulation of microRNA functions, suppressing or enhancing microRNA targeting by competitive binding on target mRNA or altering local RNA secondary structure. A key event during neuronal induction is the relief of PTB-mediated blockage of microRNA action on multiple components of the REST complex, thereby de-repressing a large array of neuronal genes, including miR-124 and multiple neuronal-specific transcription factors, in non-neuronal cells. This converts a negative feedback loop to a positive one to elicit cellular reprogramming to the neuronal lineage.

Directional movement is a property common to all cell types during development and is critical to tissue remodelling and regeneration after damage1–3. In migrating cells, calcium plays a multifunctional role in directional sensing, cytoskeleton redistribution, traction force generation, and relocation of focal adhesions1, 4–7. Here we visualise, for the first time, high-calcium microdomains (“calcium flickers”), and their patterned activation in migrating fibroblasts. Calcium flicker activity is dually coupled to membrane tension (via TRPM7, a stretch-activated Ca2+-permeant channel of the transient receptor potential superfamily8) and chemoattractant signal transduction (via type 2 inositol 1,4,5-trisphosphate receptors). Interestingly, calcium flickers are most active at the leading lamella of migrating cells, displaying a 4:1 front-to-rear polarisation opposite to the global calcium gradient6. When exposed to a PDGF gradient perpendicular to cell movement, asymmetric calcium flicker activity develops across the lamella and promotes the turning of migrating fibroblasts. These findings illustrate how the exquisite spatiotemporal organisation of calcium microdomains can orchestrate complex cellular processes such as cell migration.

Calcium (Ca 2+ ) and reactive oxygen species (ROS) constitute the most important intracellular signaling molecules participating in the regulation and integration of diverse cellular functions. Here we briefly review cross-talk between the two prominent signaling systems that finely tune the homeostasis and integrate functionality of Ca 2+ and ROS signaling systems can be both stimulatory and inhibitory, depending on the type of target proteins, the ROS species, the dose, duration of exposure, and the cell contexts. Such extensive and complex cross-talk might enhance signaling coordination and integration, whereas abnormalities in either system might propagate into the other system and undermine the stability of both systems.

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