Confounded

Remember that one batch of sand dollar larvae that were looking weird on Monday? Well, they still look weird. In fact, all of the larvae looked the same yesterday as they did on Monday, which seems strange, considering how quickly they galloped through development for the first three weeks of larval life. It's as though they've entered some stasis period during which developmental progress slows way down. Or maybe I just can't see the signs of change.

Pluteus larva of Dendraster excentricus, age 23 days. 15 April 2016 © Allison J. Gong
Pluteus larva of Dendraster excentricus, age 23 days. Mating: F2xM1. Diet: Rhodomonas only
15 April 2016
© Allison J. Gong

If I had seen these larvae for the very first time yesterday, I might not suspect that anything was strange. But having watched them twice weekly since fertilization and knowing how different they looked a week ago, my Potential Weirdness-o-Meter™ is redlining. These larvae have definitely changed in a week, and not in the way that I'm used to echinoid larvae developing. With their much shorter arms and overall stunted appearance, these guys appear to be regressing. However, they aren't dying and they don't really look bad. As I said on Monday, they just look . . . weird.

Remember how I said I'd split this cohort of larvae into two batches and fed them different things? At first I thought this strange appearance was due to the change in diet from a Rhodomonas/Dunaliella mixture to Rhodomonas only. The larva in the photo above was from the Rhodomonas-only jar, and perhaps its odd appearance could be explained by some deficiency in the monoculture diet. Then I continued on my rounds and looked at the larvae from the same mating that were still on the Rhodo/Dun diet.

Pluteus larva of D. excentricus, age 23 days. 15 April 2016 © Allison J. Gong
Pluteus larva of D. excentricus, age 23 days. Mating: F2xM1. Diet: Rhodomonas/Dunaliella mixture.
15 April 2016
© Allison J. Gong
Pluteus larvae of D. excentricus, age 23 days. 15 April 2016 © Allison J. Gong
Pluteus larvae of D. excentricus, age 23 days. Mating: F2xM1. Diet: Rhodomonas/Dunaliella mixture.
15 April 2016
© Allison J. Gong

All the larvae in these photos remained on the mixed diet, and they look pretty much the same as their siblings eating the monoculture diet. So I don't think the change in diet explains the appearance of the larvae.

Okay, then. If it's not the food that accounts for what these larvae look like, maybe it's something about the mating itself. These larvae, from both food treatments, are all full siblings from one mother mated with one father. As full sibs they share, on average, 1/4 of their DNA with each other, which could account for the similarity in their appearances. Perhaps this "strange" look is due more to genetics than to the environment (i.e., food).

I can test this hypothesis by examining larvae from the other crosses. Rather fortuitously, as it turns out, when I spawned the adult sand dollars a little over three weeks ago now, only one male contributed enough sperm for me to use. Three females spawned usable amounts of eggs, so I set up three matings:

  • F1xM1
  • F2xM1
  • F3xM1

The female designated F2 gave the most eggs, and her offspring are the ones that I split into the Rhodo-only and Rhodo/Dun diets. Note that all of the larvae in this little experiment have the same father. This gives me the opportunity to test for maternal effects on development; in other words, having controlled for the effects of different fathers--ha! I make it sound as though I did that on purpose--I can now assume that differences (in growth rate, survivability, and successful metamorphosis if we get that far) between the different matings are at least partially due to differences in egg quality among the three mothers. Or to differing gamete compatibilities between each female and the one male.

So now let's take a look at the larvae from other matings. We'll start with F1xM1:

Pluteus larva of D. excentricus, age 23 days. Mating: F1xM1. Diet: Rhodomonas/Dunaliella mix. 15 April 2016 © Allison J. Gong
Pluteus larva of D. excentricus, age 23 days. Mating: F1xM1. Diet: Rhodomonas/Dunaliella mixture.
15 April 2016
© Allison J. Gong

This larva looks normal to me, or at least what I've come to assume is normal. And wow, that was one filthy cover slip,wasn't it?

The offspring of the F3xM1 mating look very much the same:

Pluteus larva of D. excentricus, age 23 days. Mating: F3xM1. Diet: Rhodomonas/Dunaliella mixture. 15 April 2016 © Allison J. Gong
Pluteus larva of D. excentricus, age 23 days. Mating: F3xM1. Diet: Rhodomonas/Dunaliella mixture.
15 April 2016
© Allison J. Gong
Pluteus larvae of D. excentricus, age 23 days. Mating: F3xM1. Diet: Rhodomonas/Dunaliella mixture. 15 April 2016 © Allison J. Gong
Pluteus larvae of D. excentricus, age 23 days. Mating: F3xM1. Diet: Rhodomonas/Dunaliella mixture.
15 April 2016
© Allison J. Gong

And here's a short video of that same pair of larvae. They look like they're singing a duet. If I were the clever sort I'd dub in some music; alas, I'm not that clever. Does somebody want to do this for me?

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