bioRxiv Science⌕ Search

Biology subjects

Bestman, J. E.

Publications and source records attributed to Bestman, J. E..

2 recordsLinked to original sources

In vivo timelapse imaging and analysis of Golgi satellite organelle distribution and movement in the neural progenitor cells of the brain

The dividing stem cells of the developing brain are the radial glial neural progenitor cells (NPCs), multifunctional cells that proliferate to generate all of the cells of the brain, but also act as scaffolds for their migrating neuron progeny, guideposts for pathfinding growing axons and regulators of synaptic activity. These remarkable cells perform these very different activities while remaining in contact with the inner and outer surface of the ever-growing brain. NPCs synthesize proteins locally to support the compartmentalized protein expression required for the cells to perform their specialized functions, but it is not clear how the necessary processing that normally occurs in the Golgi apparatus is achieved at locations far from the cell body. Golgi satellites, motile organelles and members of the protein maturation machinery, control protein glycosylation and maturation in polarized cells like neurons. To investigate whether NPCs also rely on Golgi satellites, we expressed a fluorescent reporter to label Golgi satellites in the NPCs in the intact brains of Xenopus laevis tadpoles. Quantitative analysis of in vivo timelapse images revealed dynamic, motile Golgi satellites that distribute throughout the cell, suggesting that NPCs have local proteostasis to support their diverse functions.

neuroscience↗

In vivo time-lapse imaging of cell proliferation and differentiation in the optic tectum of Xenopus laevis tadpoles

Neural progenitor cells (NPCs) are the highly polarized dividing stem cells of the developing brain that give rise to all neurons and glia. Early on, NPCs divide symmetrically and expand the pool of progenitor cells, but as development continues the NPCs begin to asymmetrically divide to produce neurons. The mechanisms that govern this irreversible commitment to neurogenesis are not fully understood, but in other stem cell populations the regulation of mitochondria and cell metabolism is key to controlling stem cell fate. Here we use timelapse 3D confocal microscopy to observe NPCs, their cellular progeny, and their mitochondria in the developing Xenopus tectum. Our results track individual NPCs over days and show that they contain abundant mitochondria that form complicated networks distributed throughout the cells. We find that NPCs preparing to divide shift mitochondria toward the cell body where they become asymmetrically distributed, suggesting that the cells control which progeny inherit mitochondria. This uneven distribution of mitochondria in cell preparing to divide led us to test the role that mitochondria play in cell division. We overexpressed the mitochondrial biogenesis master regulator, PGC-1a, which induced the NPCs to asymmetrically divide and produce neurons, while PGC-1a knockdown limited neurogenesis. Together these data suggest that the regulation of mitochondria by NPCs prior to cell division and their unequal inheritance during cell division, contributes to the fate of the newborn cells in the developing brain.

neuroscience↗