Womb with a view

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The BBC is going to be showing a program with images of developing embryos (there are some galleries online) generated from ultrasound, cameras inserted into the uterus, and largely, computer-generated graphics. It’s all very pretty, and I hope it will also be shown in my country, but…these pictures violate all the rules of scientific imaging. The images are clearly generated by imposing artistic decisions derived from the conventions of computer animation work onto the data that was collected—I can’t tell what details in these embryos were actually imaged, and which were added by the CGI guy.

I can tell you that the way they’re rendered as free-floating individuals suspended in great airy spaces lit by a glow through distant membranes like stained glass windows is complete hokum, and the textures just look all wrong. They ought to be slimy, wrapped in membranes, and enveloped closely in maternal tissues. I hope the program includes some honest description of the process of making the images, with before and after photos, so viewers can see how much of the work is interpolated and artificially added.

MADS boxes, flower development, and evolution

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I’ve been writing a fair amount about early pattern formation in animals lately, so to do penance for my zoocentric bias, I thought I’d say a little bit about homeotic genes in plants. Homeotic genes are genes that, when mutated, can transform one body part into another—probably the best known example is antennapedia in Drosophila, which turns the fly’s antenna into a leg.

Plants also have homeotic genes, and here is a little review of flower anatomy to remind everyone of what ‘body parts’ we’re going to be talking about. The problem I’ll be pursuing is how four different, broadly defined regions of the flower develop, and what that tells us about their evolution.

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The sea urchin genome

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Oh happy day, the Sea Urchin Genome Project has reached fruition with the publication of the full sequence in last week’s issue of Science. This news has been all over the web, I know, so I’m late in getting my two cents in, but hey, I had a busy weekend, and and I had to spend a fair amount of time actually reading the papers. They didn’t just publish one mega-paper, but they had a whole section on Strongylocentrotus purpuratus, with a genomics mega-paper and articles on ecology and paleogenomics and the immune system and the transcriptome, and even a big poster of highlights of sea urchin research (but strangely, very little on echinoderm development). It was a good soaking in echinodermiana.

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Dissecting embryos from half a billion years ago

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There is a treasure trove in China: the well-preserved phosphatized embryos of the Doushantuo formation, a sampling of the developmental events in ancient metazoans between 551 and 635 million years ago. These are splendid specimens that give us a peek at some awesomely fragile organisms, and modern technology helps by giving us new tools, like x-ray computed tomography (CT), scanning electron microscopy (SEM), thin-section petrography, synchrotron X-ray tomographic microscopy (SRXTM), and computer-aided visualization, that allow us to dig into the fine detail inside these delicate specimens and display and manipulate the data. A new paper in Science describes a survey of a large collection of these embryos, probed with these new techniques, and rendered for our viewing pleasure…that is, we’ve got pretty pictures!

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Evo-devo is not the whole of biology

Sometimes a plan just comes together beautifully. I’m flying off to London tomorrow, and on the day I get back to Morris, I’m supposed to lead a class discussion on the final chapters of this book we’ve been reading, Endless Forms Most Beautiful. I will at that point have a skull full of jet-lagged, exhausted mush, and I just know it’s going to be a painful struggle. Now into my lap falls a wonderful gift.

There was a review in the NY Review of Books that said wonderful things about Carroll’s work, and in particular about the revolutionary nature of evo-devo. This prompted Jason Hodin, an evo-devo researcher himself (whose work I’ve mentioned before) to write a rebuttal and send it off to NYRB…which they chose not to publish. So he sent it to me, with permission to post it.

(If Pharyngula is going to be second choice to the NY Review of Books, I’m not going to complain.)

Anyway, I’m almost as guilty as Carroll of hawking the wares of the evo-devo bandwagon and traveling roadshow, so this is a welcome balancing corrective. The complete text is below the fold.

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Hox complexity

Here’s a prediction for you: the image below is going to appear in a lot of textbooks in the near future.

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(click for larger image)
Confocal image of septuple in situ hybridization exhibiting the spatial expression of Hox gene transcripts in a developing Drosophila embryo. Stage 11 germband extended embryo (anterior to the left) is stained for labial (lab), Deformed (Dfd), Sex combs reduced (Scr), Antennapedia (Antp), Ultrabithorax (Ubx), abdominal-A (abd-A), Abdominal-B (Abd-B). Their orthologous relationships to vertebrate Hox homology groups are indicated below each gene.

That’s a technical tour-de-force: it’s a confocal image of a Drosophila embryo, stained with 7 fluorescent probes against different Hox genes. You can clearly see how they are laid out in order from the head end (at the left) to the tail end (which extends to the right, and then jackknifes over the top). Canonically, that order of expression along the body axis corresponds to the order of the genes in a cluster on the DNA, a property called colinearity. I’ve recently described work that shows that, in some organisms, colinearity breaks down. That colinearity seems to be a consequence of a primitive pattern of regulation that coupled the timing of development to the spatial arrangements of the tissues, and many organisms have evolved more sophisticated control of these patterning genes, making the old regulators obsolete…and allowing the clusters to break up without extreme consequences to the animal. A new review in Science by Lemons and McGinnis that surveys Hox gene clusters in different lineages shows that the control of the Hox genes is much, much more complicated than previously thought.

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Cellular responses to alcohol

Forgive me, but I’ll inflict a few more zebrafish videos on you. YouTube makes this fun and easy, and I’m going to be giving my students instruction in video micrography next week, so it’s good practice.

This is a more detailed look at what’s going on in the embryo. Using a 40x objective, we zoom in on a patch of cells near the surface of a 4-hour-old embryo—this is a generic tissue called the blastoderm. We just record activity with an 1800-fold time compression for a few hours to see what the cells are doing. The movie below displays typical, baseline activity: the cells are jostling about, you’ll see an occasional mitosis, and sometimes you’ll see a cell vanish out of focus as it moves deeper into the embryo, and sometimes you’ll suddenly see a new cell squirm to the surface. It’s all just a happy, dynamic place with lots of random motion; these can be mesmerizing to watch.

These blastoderm sheets are a kind of cellular testbed for quick assays of the effects of teratogens on embryonic tissues. We just wash the embryo with whatever substance we’re interested in testing, and see if and how the cells react.

Alcohol is a dramatic example. Here’s a blastoderm sheet under stress as it is exposed to 3% ethanol.

Some obvious changes are going on. One is that the surfaces of the individual cells are seething—they are bubbling out and sucking back in little balloons of membrane, a process called blebbing. This is a very typical response to any kind of stress. Apparently, mitosis is another kind of stress: we can reduce the concentration of alcohol so that the cells look normal, except that as they’re about to divide they go into a flurry of blebbing that persists until division is complete.

We had another puzzle to solve. Sometimes, as we were looking at our low magnification recordings of embryos, we’d see the whole blastula or gastrula shudder. They don’t have muscles yet! We didn’t know what was causing pulses of contractile activity to sweep across the whole animal at such a relatively undifferentiated stage.

These movies show what was going on. They’re a real pain to keep in focus, because in addition to the fine blebbing activity in individual cells, the whole surface occasionally dimples and changes shape. What’s happening? Cells are dying somewhat randomly, some on the surface, some deeper in the embryo. Deep cells that die seem to be actively evicted from interior; sometimes the surface will buckle inward (with the image going out of focus), and when it bounces back up, it ejects a load of cellular debris out into the external medium. There’s a particularly dramatic example at the end of this movie, where everything in the lower half goes massively out of focus, and when it bounces back, it carries a large dead cell that sits there briefly, then abruptly pops and disappears.

If you look at that earlier lower resolution movie of ethanol effects, you might notice odd rough blobs on the surface of the embryo, and we think what that is is the extruded debris of deep cells killed by alcohol exposure, thrown up out of the interior to prevent them from interfering with normal development. This is actually a rather cool cellular mechanism that helps embryos survive random glitches in the process of building these massive pools of cells as it grows—it’s a kind of tissue-level garbage disposal service.

Developing under the influence: zebrafish in alcohol

Ah, the evils of strong drink. Or weak drink. You all know that you shouldn’t drink alcohol to excess during pregnancy, and the reason is that it can affect fetal development. We take zebrafish eggs and put them on a real bender: we soak them in various concentrations of alcohol (which are hard to compare with human blood alcohol levels, I’m afraid, but trust me: these are such gross levels of ethanol that mere humans would be dead and pickled. Fish are tough), and let them stew for hours. Since fish development is much, much faster than human development, it’s rather like having a woman start drinking straight Everclear a few weeks after discovering she’s pregnant, and staying snockered throughout the first trimester.

So don’t try this at home, kids.

The animal on the left is a teetotaler control. The one on the right is going to get washed in 3% alcohol at about 4 hours of development. It’ll be obvious; a label will pop up, and also the eggs are embedded in agar to immobilize them, and the agar will go cloudy and dark for a while as the alcohol soaks in.

Even if you aren’t intimately familiar with fish embryology, you should be able to see that the one on the right develops more slowly. Especially at the end, the one on the left will be twitching vigorously and spinning in the chorion, while the lush on the right is much slower. There are also some subtle deformities in tail shape, and you might notice odd schmutzy gunk on the animal’s epidermis…more about that later.

Also, you’ll notice that we started both recordings immediately after fertilization—I was hovering over the tank, and as soon as momma and daddy squirted out the gametes, I scooped them up and slapped ’em down in a dish, to guarantee that everything was starting precisely in synchrony. These movies start a little earlier and go on a little longer than the previous example.

That zebrafish movie annotated and explained, a little

By popular request, here’s a roughly annotated version of that zebrafish development movie.

Stuff to watch for:

  • This movie starts at the 8-16 cell stage. The cells of the embryo proper (blastomeres) are at the top, sitting on a large yolk cell.

  • The pulsing is caused by the synchronous early divisions of all the cells. They lose synchrony at the mid-blastula transition.

  • Epiboly is the process by which the cells migrate downward over the yolk. An arrow will briefly flash, pointing to about 11:00, in the direction of the animal pole (where the future nose will form, sorta). That happens just before the whole animal begins to rotate within the chorion, just to make following everything more difficult.

  • After the animal rolls over, the animal pole is pointing straight up at you, and the migrating cells will form the germ ring, a thickening around the equator of the embryo. Cells will also migrate towards one point along the ring, forming a thickening called the keel. This is where the embryonic axis is forming; cells are migrating into the interior at this point in the process called gastrulation, and this region is roughly equivalent to the dorsal lip of a frog.

  • The whole animal is going to roll over again, this time to its side. The keel is thickening and lengthening towards the animal pole. The body of the fish is going to form along the right side of spherical embryo in this view.

  • While the keel is extending anteriorly, cells are still also migrating to surround the yolk—epiboly continues, with the yolk bulging out a bit until it is finally surrounded and closed off at the blastopore.

  • The head and tail extend. You’ll see the eye forming, so you’ll be able to tell which end is the head end.

  • Along the right side, you’ll also see the tissue form regular little blocks: these are the somites, or body segments.

  • The tail continues to extend and lifts off the surface of the yolk. When there are about 18 somites (the resolution is too low, so don’t try to count them), the animal will begin to twitch.

I’ll load up another one in a bit that will show a hint of the horrible stuff we do to them in the lab: we get the babies drunk and watch deformities develop.