December 18, 2014



You might not want the dreaded tube socks in your Christmas stocking this year, but you do appreciate the actual tubes that your body depends on in just about every organ system. A recent paper in PLOS Biology describes tube formation in the fly renal system and the signals that regulate it.

Tubes generally start as buds that dramatically elongate during development, but the cell rearrangements that occur during tubulogenesis are not completely understood. Saxena and colleagues recently used the developing fly renal system to track cell movements during tube formation. Tubule elongation primarily occurs through convergent extension, during which cells intercalate along the length of the tube. During these rearrangements, the number of cells around the circumference of the tube drops as the number of cells along the tube increases. Saxena and colleagues show that epidermal growth factor localized at the tip cells of the distal end of the tube guides the polarity of cell rearrangements, via polarization of Myosin II within individual cells. Finally, without proper tube elongation, animals have abnormal excretory function and osmoregulation, leading to lethality. In the images above, the top row shows failure of tube elongation after laser ablation of the distal tip cells (arrowheads). Bottom row shows normal tube elongation without laser ablation of tip cells (arrowheads).

Saxena, A., Denholm, B., Bunt, S., Bischoff, M., VijayRaghavan, K., & Skaer, H. (2014). Epidermal Growth Factor Signalling Controls Myosin II Planar Polarity to Orchestrate Convergent Extension Movements during Drosophila Tubulogenesis PLoS Biology, 12 (12) DOI: 10.1371/journal.pbio.1002013

November 26, 2014

 
Patterns are soothing for left-brained folks like me, with the exception being those terrible patterned holiday sweaters that will come out of mothball-ridden closets soon (unsettling for everyone, really). Today’s images are from a paper describing a new micropatterning technique to look at plasma membrane proteins. 

The plasma membrane of a cell is riddled with many multi-protein complexes that facilitate communication and transport. These complexes provide a challenge to biologists due to their ubiquitous localization around the cell, their large, complex size, and their transient interactions with proteins. A recent paper describes a technique to study signaling complexes at the plasma membrane, using micropatterns within the plasma membrane. Löchte and colleagues expressed a protein bait in a micropattern in a living cell’s plasma membrane. Dynamics of the interactions between the protein bait and ligand target can then be quantified using live microscopy and on a single-molecule level. Löchte and colleagues used the IFN interferon signaling complex, using one of the receptor units (IFNAR2) as bait. In the top images above, the micropatterned receptor bait (IFNAR2) was able to recruit the other IFN receptor subunit (IFNAR1, green) after the addition of the ligand (red; time represents the addition of the ligand). Bottom image shows a closer view of the high-affinity, micropatterned binding that requires simultaneous interaction of the ligand (red) with both receptor subunits.


Lochte, S., Waichman, S., Beutel, O., You, C., & Piehler, J. (2014). Live cell micropatterning reveals the dynamics of signaling complexes at the plasma membrane originally published in the Journal of Cell Biology, 207 (3), 407-418 DOI: 10.1083/jcb.201406032

November 19, 2014

You might not be able to get rid of the bad guys, but you can still win the battle if you cripple their mobility. Today’s image is from a paper describing how a tumor’s microenvironment can predict the motility of individual tumor cells.

Metastasis is the spread of cancer cells throughout the body. The motility of tumor cells depends on the microenvironment around them, and a recent paper systematically looks at how that microenvironment can predict or alter the behavior of tumor cells. Gligorijevic and colleagues tracked the motility of individual mouse mammary carcinoma cells in vivo using high-resolution multi-photon microscopy, and found that tumor cells exhibited either fast or slow locomotion. Those tumor cells with slow locomotion also exhibited invadopodia, protrusions that Gligorijevic and colleagues directly link to degradation of the underlying extracellular matrix and metastasis. While no single parameter of the tumor’s microenvironment could predict the locomotion of tumor cells, a support vector machine algorithm indicated how combinations of many parameters could predict tumor cell phenotype and behavior. By characterizing the heterogeneous microenvironment of a tumor and predicting the location and behavior of metastatic tumor cells, researchers can better understand treatment of tumors and the varying responses. Images above show protrusions (arrowheads, over 30 minutes) on two different tumor cells with slow locomotion, with protrusions facing collagen fibers (purple).

BONUS!! Check out a movie of these protrusions below. Note that some protrusion face collagen fibers (purple, panels a and b), and some protrude into blood vessels (red, panels c and d).
video

BONUS!! Check out Bojana Gligorijevic ‘s interview with SciArt about her images, research, and art here.

Gligorijevic, B., Bergman, A., & Condeelis, J. (2014). Multiparametric Classification Links Tumor Microenvironments with Tumor Cell Phenotype PLoS Biology, 12 (11) DOI: 10.1371/journal.pbio.1001995

November 12, 2014

You might think that the “kiss-and-hop” is a dance move strictly forbidden at a Duggar homeschool prom, but it refers to the quick dynamics of the microtubule-associated protein tau. Today’s image is from a paper describing unexpected results about how tau resides on and regulate microtubules without physically impeding microtubule motors. 

The microtubule-associate protein tau binds to and stabilizes the microtubules within an axon. As most tau is believed to decorate axonal microtubules, it was previously unclear how tau can function in its non-microtubule-dependent roles, or how the presence of tau does not interfere with microtubule motors and axonal transport. A recent paper by Janning and colleagues describes the use of single-molecule tracking of tau in living cells. Janning and colleagues found that tau resides on a single microtubule for 40ms before hopping to the next microtubule, and that this unexpectedly short residence time is sufficient to affect microtubule stability. This “kiss-and-hop” mechanism allows for normal axonal transport, and suggests how some tau functions away from microtubules. In the images above, tau (red) is labeled in mouse cortical neurons, as are microtubules (green) and the nucleus (blue).  

Janning, D., Igaev, M., Sundermann, F., Bruhmann, J., Beutel, O., Heinisch, J., Bakota, L., Piehler, J., Junge, W., & Brandt, R. (2014). Single-molecule tracking of tau reveals fast kiss-and-hop interaction with microtubules in living neurons Molecular Biology of the Cell, 25 (22), 3541-3551 DOI: 10.1091/mbc.E14-06-1099

October 28, 2014

If you’ve ever tried to get your kids to share a donut, you understand the importance to dividing things equally (and learning crucial lessons…just buy more donuts next time...I mean, seriously!). Cell division is no different—chromosomes and organelles must all get divided equally. Today’s images are from a paper showing how mitochondria are positioned during cell division in order to allow equal segregation.

Many years of research have focused on the equal segregation of chromosomes during cell division. Organelles such as mitochondria must also be segregated equally in a dividing cell, and errors in this process can lead to disease. A recent paper identifies the actin motor Myosin-XIX (Myo19) as a key player in mitochondrial partitioning during cell division. Myo19 is localized to mitochondria, and cells depleted of Myo19 have an abnormal distribution of mitochondria. Rohn and colleagues found that cells lacking Myo19 experience stochastic division failure, suggesting that mitochondria are physically preventing successful cell division. The images above show dividing cells labeled to visualize mitochondria (green) and the mitotic spindle (red) in control cells (top two rows) and cells depleted of Myo19 (bottom two rows). Without Myo19, mitochondria moved towards spindle poles at the onset of anaphase, causing an asymmetric distribution at division when compared with control cells.

BONUS!! Here is a rotating 3D reconstruction of an A549 stained to visualize microtubules (green), mitochondria (red), and DNA (blue). Omar Quintero, HighMag friend and a co-author from today’s paper, loves this image: “I like it because it reminds me of the scenes in StarWars where the Rebels are planning their attack on the Death Star.”
video

Rohn, J., Patel, J., Neumann, B., Bulkescher, J., Mchedlishvili, N., McMullan, R., Quintero, O., Ellenberg, J., & Baum, B. (2014). Myo19 Ensures Symmetric Partitioning of Mitochondria and Coupling of Mitochondrial Segregation to Cell Division Current Biology DOI: 10.1016/j.cub.2014.09.045

Copyright ©2014 Elsevier Ltd. All rights reserved.

October 24, 2014

There is a party going on at the ends of microtubules, but I wasn’t invited. That won’t stop me, or countless cell biologists out there, from peeping in the window to check out all of the microtubule shenanigans. Today’s image is from a paper describing how Doublecortin binds to microtubule ends.

The plus end of a microtubule is the primary site for growth and shrinkage, and interaction with several microtubule-associate proteins. Different microtubule end-binding proteins may interact with microtubules using different mechanisms: the end-binding protein EB1 relies on the nucleotide state of the tubulin at the microtubule end, while a recent paper shows how another protein, Doublecortin (DCX), relies on the curvature of microtubule ends for binding. DCX is a neuronal microtubule-associate protein that plays an important role throughout development, yet how it interacted with microtubule ends was previously unclear. Bechstedt and colleagues used single-molecule microscopy to show that DCX (images above, green in merged) binds with higher affinity to curved microtubules (magenta) than to straight microtubules. DCX mutations, which are found in patients with double cortex syndrome, prevent the protein from binding to curved regions of microtubules.

Bechstedt, S., Lu, K., & Brouhard, G. (2014). Doublecortin Recognizes the Longitudinal Curvature of the Microtubule End and Lattice Current Biology, 24 (20), 2366-2375 DOI: 10.1016/j.cub.2014.08.039
Copyright ©2014 Elsevier Ltd. All rights reserved.


October 17, 2014

For years, the prettiest cells to image were flat cells in a dish. Thanks to the tireless work of many, beautiful high-resolution images can now come from tissue within a living organism. Today’s image is from a paper showing improved techniques for imaging fine cellular processes within large volumes, from the lab of recent Nobel prize winner, Eric Betzig. 

A material’s refractive index refers to how light travels through it; the simplest example being how light bends when passed through water. The refractive index heterogeneities stemming from the many cell types, morphologies, and subdomains within a living organism are a challenge to microscopists. As described in a paper from earlier this year, Wang and colleagues improved on previous techniques for imaging within large volumes. Wang and colleagues use adaptive optics (AO), which corrects for the microscope’s aberrations that limit image resolution, in a mode fast enough to correct for the various aberrations within a large sample, without inducing photodamage or photobleaching. The image above shows a 3D rendering from deep within a living zebrafish brain, with oligodendrocytes (magenta) and neuronal nuclei (green) visible.

Wang, K., Milkie, D., Saxena, A., Engerer, P., Misgeld, T., Bronner, M., Mumm, J., & Betzig, E. (2014). Rapid adaptive optical recovery of optimal resolution over large volumes Nature Methods, 11 (6), 625-628 DOI: 10.1038/nmeth.2925
Adapted by permission from Macmillan Publishers Ltd, copyright ©2014