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Strangeness abounds

Wow, they weren't kidding about "early developmental asynchrony" in sea stars! This morning I looked at the embryos that I had started almost 24 hours earlier, and noticed two things right off the bat:

Thing #1:  Within the F1 x M1 (Purple female x Purple male) mating , developmental rates among full siblings were all over the map. Some embryos had progressed to the blastula stage, which is essentially a hollow ball of ciliated cells, while others were still in the early cleavage stages and rather a lot hadn't divided at all. In fact, with 24 hours of hindsight I can see that several of these eggs had not even been fertilized.

Embryos of Pisaster ochraceus, age 24 hrs. 3 June 2015. © Allison J. Gong
Embryos of Pisaster ochraceus, age 24 hrs
3 June 2015
© Allison J. Gong

My first reaction upon looking into the microscope and seeing all these assorted blobs was, "Oh, crap." But then I looked more closely at some of the embryos and realized that they had become blastulae!

Here's a picture of a blastula. This embryo is freely swimming inside its fertilization envelope, although it doesn't have a lot of space (remember that narrow perivitelline space from yesterday? that's all the elbow room it has). The hollow space in the center of the embryo is the blastocoel 'sprout cavity.' Given that the embryo hasn't grown (or even hatched) yet, it's still ~165 µm in diameter, the size of the original egg.

Blastula of Pisaster ochraceus, 3 June 2015. © Allison J. Gong
Blastula of Pisaster ochraceus
3 June 2015
© Allison J. Gong

The stage that precedes the blastula (a hollow ball of cells) is called a morula (a solid ball of cells). The embryo that is partially visible in the bottom of the above photo may be a morula. Imagine the following sequence of events: (1) an egg is fertilized by a sperm, forming a zygote; (2) the zygote undergoes a number of cleavage divisions, with the cells becoming more numerous and smaller in size; (3) at some stage a solid ball of small cells, the morula, is formed; (4) as cell division continues, the cells migrate toward the outside of the sphere, forming a cavity (the blastocoel) in the middle.

The blastula is a ciliated stage, and in this video clip you can see the cilia moving. I shot this video at only 100X magnification to capture as much depth of field as possible, and suggest viewing at full-screen. This should enable you to see the three-dimensional structure of the embryo, and that it is indeed a sphere.

Thing #2:  The F2 x M1 mating (Orange female X Purple male) isn't doing well at all. I looked at several slides and didn't see any embryos that were developing normally. They had all been fertilized, as I could see the fertilization envelope surrounding each egg, but most had not even divided. The ones that had divided were all strange and just plain wrong. Here, see for yourself:

24-hr embryos of Pisaster ochraceus, 3 June 2015. © Allison J. Gong
24-hr embryos of Pisaster ochraceus
3 June 2015
© Allison J. Gong

Many of the eggs are blurry because they're below the focal plane of the microscope. But see how many of them are undeveloped? And how, in the ones that have started dividing, the cells are disorganized and of different sizes? Typical echinoderm cleavage, as I see in echinoids (our local urchins and sand dollars) and in my other crossing of these ochre stars, results in a blastula made up of cells that are all approximately the same size. Most of these embryos, on the other hand, appear to consist of one large cell and a bunch of tiny ones.

I assume that these abnormal-looking-to-me embryos will not hatch, although I could be pleasantly surprised tomorrow. I don't yet have much of an intuition about these Pisaster ochraceus embryos, so this is a huge learning experience for me. I do expect to see hatching in the F1 x M1 cross tomorrow. Fingers crossed!

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