August 19, 2014

Think of life without tubes for a moment. Not only would our huge bodies cease to exist, but our homes’ plumbing would be a mess and my 5-year old’s marble run would be pretty boring. The formation of tubes during development is a fascinating topic. Today’s image is from a paper describing the role of endocytosis in seamless tube formation.

The trachea of the fruit fly is a simple tubular system that functions as the respiratory system of the fly. The star-shaped tracheal terminal cells form seamless tubes that extend the length of long cellular extensions. Schottenfeld-Roames and colleagues recently published a study investigating the mutations in the braided gene. Tracheal terminal cells in braided mutants have tubular cysts and fewer branches, as seen in the images above (top is wild-type; bottom is mutant). braided encodes Syntaxin7, a endocytosis protein that promotes fusion of vesicles to early endosomes. Schottenfeld-Roames and colleagues found that mutations in other early endosome proteins cause similar terminal cell tube defects. Additional data showing increased levels of the apical protein Crumbs in braided terminal cells suggests that early endocytosis regulates levels of Crumbs, which in turn affects tube formation through actin cytoskeleton modulation. The images above show the luminal membrane (green) and an apical protein (magenta) in tracheal tubes. The tubes in braided mutants are cystic and abnormal, and the tube tips are disorganized (higher magnified views of the boxed regions are on the left).

Schottenfeld-Roames, J., Rosa, J., & Ghabrial, A. (2014). Seamless Tube Shape Is Constrained by Endocytosis-Dependent Regulation of Active Moesin Current Biology, 24 (15), 1756-1764 DOI: 10.1016/j.cub.2014.06.029
Copyright ©2014 Elsevier Ltd. All rights reserved.

All the images were acquired by Dr. Jodi Schottenfeld-Roames.

August 14, 2014

Astrocytes used to be the red-headed stepchild of the neurobiology world, but no more! Once considered to be just filler material, astrocytes are now known to function in the development and function of synapses, though the mechanisms are unclear. Today’s stunning image is from a paper showing how astrocytes can stabilize synapses, possibly serving as an important component of learning and memory. 

The synapses of neurons in the central nervous system are dynamic in response to learning and memory. The synapses are enveloped by perisynaptic astrocytic processes (PAPs), which are intricate processes of astrocytes. This close association of PAPs with synapses suggests an important role for astrocytes in synaptic development, transmission, and plasticity—the focus of a recent paper by Bernardinelli and colleagues. In this study, time-lapse imaging of brain slices revealed that long-term potentiation increased PAP motility and astrocyte coverage of the synapse. In vivo imaging of the somatosensory cortex of adult mice after whisker stimulation showed an increase in PAP motility, and later dendritic spine stability. From these results, Bernardinelli and colleagues identify a novel bidirectional interaction between PAPs and synapses, in which synaptic activity regulates PAP plasticity, which in turn regulates PAP coverage of synapses and long-term spine survival. The image above shows CA1 neurons (green) and stratum radiatum astroctyes (red) in mouse hippocampal tissue.

Bernardinelli, Y., Randall, J., Janett, E., Nikonenko, I., König, S., Jones, E., Flores, C., Murai, K., Bochet, C., Holtmaat, A., & Muller, D. (2014). Activity-Dependent Structural Plasticity of Perisynaptic Astrocytic Domains Promotes Excitatory Synapse Stability Current Biology, 24 (15), 1679-1688 DOI: 10.1016/j.cub.2014.06.025
Copyright ©2014 Elsevier Ltd. All rights reserved.

August 7, 2014

No matter how many brilliant discoveries are made by countless brilliant scientists, there will always be a lot of unknowns out there. These unknowns are what keep our mental wheels turning, our imaginations running, and our labs busy. Today’s image is from a paper that describes a newly-discovered process of vascular development called “canalogenesis.”

Schlemm’s canal (SC) is a flattened tube that encircles the anterior portion of the eye and drains fluid from the area. Abnormal drainage contributes to glaucoma, a disease that causes vision loss, yet a description of SC development and SC endothelial cells (SECs) is incomplete. In a recent study, Kizhatil and colleagues developed a new whole-mount procedure and used high-resolution confocal microscopy to study large regions of the SC during development. Kizhatil and colleagues found that the phenotype of SECs is a blend of blood and lymphatic endothelial cells, and that the SC develops through by a newly-discovered process called “canalogenesis.” Canalogenesis has features that are similar to, yet different from, the three well-studied vascular development programs—vasculogenesis, angiogenesis, and lymphangiogenesis. The image above was acquired using the new whole-mount procedure that protects the delicate ocular drainage structures. The SC (blue) is visualized in 3D relative to nearby blood vessels (magenta).

Kizhatil, K., Ryan, M., Marchant, J., Henrich, S., & John, S. (2014). Schlemm's Canal Is a Unique Vessel with a Combination of Blood Vascular and Lymphatic Phenotypes that Forms by a Novel Developmental Process PLoS Biology, 12 (7) DOI: 10.1371/journal.pbio.1001912

July 31, 2014

Do you ever feel nostalgic for a specific paper? Maybe this paper inspired your own research, or maybe it was a paper you immediately knew would be game-changing. Maybe, like today’s TBT paper, it was a great paper about solidly executed research with a memorable giggle-inducing technique. Thanks to a nostalgic HighMag reader and friend, Omar Quintero, we are being re-introduced to gonad sandwiches. 

In mammals, sex determination refers to the changes during early development that lead to the formation of either the testis or ovary. A gene on the Y chromosome called Sry initiates testis formation from the early bipotential gonad, including organizing Sertoli cells into the testis cord structure. In a 1997 paper, Martineau and colleagues investigated the early cell movements that occur after Sry expression, specifically the movement of nearby mesonephric cells to the genital ridge. To see these cell movements, Martineau and colleagues grafted a “blue” mesonephros from a mouse ubiquitously expressing β-galactosidase next to a “white” gonad from a different mouse. The movement of blue cells into the white gonad in these gonad sandwiches revealed that this movement is dependent on a signal induced by the male (XY) gonad that acts as a chemoattractant. Migration does not occur if an XX gonad is used in the sandwich, yet migration can occur whether an XY or XX mesonophros is used. The images above show the different XX and XY combinations used in these experiments, with XY gonads leading to extensive migration of blue cells. 

Martineau, J., Nordqvist, K., Tilmann, C., Lovell-Badge, R., & Capel, B. (1997). Male-specific cell migration into the developing gonad Current Biology, 7 (12), 958-968 DOI: 10.1016/S0960-9822(06)00415-5

Copyright ©1997 Elsevier Ltd. All rights reserved.

July 24, 2014

How many times can you say the word “gonad” in a sentence without giggling? If the answer is none, then I congratulate you on turning thirteen. If the answer is many, then you must be a biologist. Biologists appreciate the value of a good gonad, and so should you. The gonad of the worm C. elegans serves as an important model in which to study tissue organization and development, as you’ll see in the paper that accompanies today’s image.

At the end of cell division, cytokinesis typically results in two separate daughter cells. Some cytokinesis, though, is incomplete and leads to two daughter cells sharing cytoplasm. This shared cytoplasm, or syncytium, can be found in the germ cells of many species from worms to humans. The germline of the worm C. elegans is a polarized tube in which germ cells are arranged around the shared cytoplasmic core and move along a conveyer belt of oocyte production. Amini and colleagues recently reported on the formation of the syncytial C. elegans germline throughout development, and the role of the short Anillin family scaffold protein ANI-2. ANI-2 is localized to the intercellular bridges that connect the germ cells to the shared cytoplasm, and loss of ANI-2 results in destabilization of intercellular bridges and sterility. The defects seen in worms lacking ANI-2 are likely due to a loss of the stability and elasticity of the intercellular bridges that is required to compensate for the stress of cytoplasmic streaming during oogenesis. Images above show the germlines of wild-type and ani-2(-) worms at different larval stages (membranes in green; nuclei in red). Worms lacking ANI-2 have abnormal multinucleated germ cells (arrowheads).

Amini, R., Goupil, E., Labella, S., Zetka, M., Maddox, A., Labbe, J., & Chartier, N. (2014). C. elegans Anillin proteins regulate intercellular bridge stability and germline syncytial organization originally published in the Journal of Cell Biology, 206 (1), 129-143 DOI: 10.1083/jcb.201310117

July 18, 2014

Poor polar bodies typically go the way of that old container of Chinese take-out in your fridge and are eventually dumped. Thanks to a very clever study published in Cell, polar body transfer can prevent the transmission of inherited mitochondrial diseases. Waste not, want not.

The meiotic divisions of an oocyte result in the production of an egg in the extrusion of two very small polar bodies. These polar bodies have the same genetic material as the egg but have only a small number of organelles, including mitochondria. The DNA of mitochondria (mtDNA) can carry mutations that cause a variety of diseases. As mtDNA is maternally inherited due to the abundance of mitochondria in the oocyte, recent studies have focused on the replacement of mutant mtDNA with normal mitochondria to treat these inherited diseases. A recent paper tests the use of polar bodies as the source of donor genomes in a potential new method for mitochondrial replacement. As polar bodies have the same genome as the egg, but does not have mtDNA variants, they can successfully replace the genome in a recipient egg that already has normal mtDNA. Wang and colleagues have shown that polar body genome transfer successfully does just this, and provides a potential new therapy for preventing inherited mitochondrial diseases. The images above show the presence of mitochondria (red) in oocytes and polar bodies. Both polar bodies (PB1 and PB2) have far fewer mitochondria than the ooctyes.

Wang, T., Sha, H., Ji, D., Zhang, H., Chen, D., Cao, Y., & Zhu, J. (2014). Polar Body Genome Transfer for Preventing the Transmission of Inherited Mitochondrial Diseases Cell, 157 (7), 1591-1604 DOI: 10.1016/j.cell.2014.04.042
Copyright ©2014 Elsevier Ltd. All rights reserved.

July 10, 2014

Do your thoughts and feelings have colors? Do you feel red with rage during traffic, or green with envy when your lady swoons over Ryan Gosling? A recent methods paper introduces a very cool technique that allows the visualization and measurement of voltage within an excited neuron.

Biologists build tools that are ideally accurate, fast, and non-damaging to the cells and organisms on which they are used. In a recent paper in Nature Methods, Hochbaum and colleagues describe the improved technique for simultaneous imaging of neuron stimulation and the resulting action potentials. Hochbaum and colleagues engineered a vector, called Optopatch, that uses their actuator (CheRiff) to induce action potentials and their voltage indicators (QuasAr1 and QuasAr2) to visualize and measure membrane voltage. Optopatch allows the measurement of action potentials on a microsecond timescale, without the need for electrodes. In the images above, a neuron expressing Optopatch shows action potential propagation (left to right, arrow is site of action potential initiation).

Hochbaum, D., Zhao, Y., Farhi, S., Klapoetke, N., Werley, C., Kapoor, V., Zou, P., Kralj, J., Maclaurin, D., Smedemark-Margulies, N., Saulnier, J., Boulting, G., Straub, C., Cho, Y., Melkonian, M., Wong, G., Harrison, D., Murthy, V., Sabatini, B., Boyden, E., Campbell, R., & Cohen, A. (2014). All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins Nature Methods DOI: 10.1038/nmeth.3000
Adapted by permission from Macmillan Publishers Ltd, copyright ©2014