Mü ller glia can serve as a source of new neurons after retinal damage in both fish and birds. Investigations of regeneration in the mammalian retina in vitro have provided some evidence that Mü ller glia can proliferate after retinal damage and generate new rods; however, the evidence that this occurs in vivo is not conclusive. We have investigated whether Mü ller glia have the potential to generate neurons in the mouse retina in vivo by eliminating ganglion and amacrine cells with intraocular NMDA injections and stimulating Mü ller glial to re-enter the mitotic cycle by treatment with specific growth factors. The proliferating Mü ller glia dedifferentiate and a subset of these cells differentiated into amacrine cells, as defined by the expression of amacrine cell-specific markers Calretinin, NeuN, Prox1, and GAD67-GFP. These results show for the first time that the mammalian retina has the potential to regenerate inner retinal neurons in vivo.amacrine ͉ Gabaergic neuron ͉ glia I t is well established that the retina of cold-blooded vertebrates regenerates very well after damage (1-3). The avian retina is also capable of limited regeneration of new neurons following neurotoxic damage (4). In both systems, damage to retinal neurons causes Müller glial cells to re-enter the cell cycle, after which they dedifferentiate into retinal progenitors, and ultimately differentiate into neurons. In fish, all types of neurons are regenerated. However, in chicks, only a limited number of different types of inner retinal neurons (amacrine, bipolar, and ganglion cells) are produced; few, if any, photoreceptors are regenerated.In the mammalian retina, by contrast, the proliferative response of Müller glia to injury is even more limited than in chicks. In response to injury in mouse or rat retina, the Müller glia may become reactive and hypertrophy, but few re-enter the mitotic cell cycle. Due to the lack of a spontaneous regenerative response in the mammalian retina, several groups have attempted to stimulate regeneration with intraocular injections of growth factors and/or transcription factors, in vitro or in vivo (5-10). Taken together, the studies of the mammalian retina indicate that Müller glia have a very limited proliferative response to injury, but can be stimulated to re-enter the cell cycle after photoreceptor or inner retinal neuron injury. These studies also reported that some of the progeny of Müller glial mitotic divisions go on to differentiate characteristics of rod photoreceptors. However, these studies failed to detect regenerating inner retinal neurons after damage, unless they were transfected with genes that specifically promote amacrine fate (9). This is puzzling, since in the chick, amacrine cells are the primary neuronal cells that are regenerated after injury. In an attempt to resolve this issue, we have carried out a systematic analysis of the response to injury in the mouse retina, and the effects of growth factor stimulation on Müller glial proliferation. The previous studies have used rats because ...

Ferguson analysis of protein electromigration from single-cell lysate in an open microfluidic device to inform optimal assay design.

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Diazirine-functionalized hydrogels have lower fluorescence background signal compared to benzophenone-functionalized hydrogels for in-gel immunoassays and can improve the lower limit of detection of fluorescence immunoassays.