Direct visualization of cell division using high-resolution imaging of M-phase of the cell cycle

Abstract

Current approaches to monitor and quantify cell division in live cells, and reliably distinguish between acytokinesis and endoreduplication, are limited and complicate determination of stem cell pool identities. Here we overcome these limitations by generating an in vivo reporter system using the scaffolding protein anillin fused to enhanced green fluorescent protein, to provide high spatiotemporal resolution of mitotic phase. This approach visualizes cytokinesis and midbody formation as hallmarks of expansion of stem and somatic cells, and enables distinction from cell cycle variations. High-resolution microscopy in embryonic heart and brain tissues of enhanced green fluorescent protein–anillin transgenic mice allows live monitoring of cell division and quantitation of cell cycle kinetics. Analysis of cell division in hearts post injury shows that border zone cardiomyocytes in the infarct respond with increasing ploidy, but not cell division. Thus, the enhanced green fluorescent protein–anillin system enables monitoring and measurement of cell division in vivo and markedly simplifies in vitro analysis in fixed cells.

Retinal pigment epithelial cell multinucleation in the aging eye - a mechanism to repair damage and maintain homoeostasis

“…2C,D). These giant RPE cells stained diffusely with propidium iodide, a sign of cell death, and lacked evidence of definitive nuclei (Hesse et al ., 2012) (Fig. 2C,D).…”

Asymmetric inheritance of the apical domain and self-renewal of retinal ganglion cell progenitors depend on Anillin function

“…Anillin is an established midbody component, and asymmetric inheritance of the midbody during stem cell division has been reported in multiple systems, including mouse radial glial progenitors (Dubreuil et al, 2007;Ettinger et al, 2011;Hesse et al, 2012;Kosodo et al, 2004). This led us to investigate Anillin-eGFP dynamics in dividing retinal progenitor cells.…”

Exosomes and Cardiac Repair After Myocardial Infarction

“…5,8,9 Because of the heart's limited ability to regenerate rapidly after a catastrophic insult, such as a MI, scar formation, rather than muscle regeneration, is often a major component of the healing response. The regeneration/proliferation of cardiomyocytes may be faster in the border zone of an injured heart as compared with the normal heart, 10 both in mice 11 and in humans. 5 Nevertheless, the limited endogenous reparative mechanisms in the adult mammalian heart seem to depend more on the replenishment by progenitor cells than on replacement by cardiomyocyte proliferation.…”