…and this is a 
Developing under the influence: zebrafish in alcohol
Category: Development • Organisms • Science
Posted on: September 28, 2006 5:39 PM, by PZ Myers
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.
Comments
This is your zebrafish on drugs.
Posted by: QrazyQat | September 28, 2006 5:53 PM
so... what exactly does this tell you? Do the pickled fish perform poorly on whatever fish intelligence test you have?
Posted by: zwa | September 28, 2006 6:18 PM
So don't try this at home, kids.
Already tried. The sea monkeys did not survive. :(
Posted by: Grumpy | September 28, 2006 6:31 PM
That is just too cool. Thanks a lot for sharing. Call me greedy, but I want more!
Posted by: Dave Carlson | September 28, 2006 6:36 PM
I was going to email this but since you're talking about getting fish drunk I'll post it here...what happens when you get a frog drunk and take it scuba diving?
http://www.rsc.org/Publishing/ChemicalBiology/Volume/2006/10/Anaesthetisedbrainsunderpressure.asp
Posted by: Aa | September 28, 2006 6:44 PM
Very nifty! When I did the same experiments (with 2% ethanol), my zebrafishies developed cardiac edemas (big balloons of fluid on the heart)! Is that supposed to happen?
Posted by: Cat | September 28, 2006 8:51 PM
Even without the drunk fish on the right, it's just stunningly cool to see the development of the embryo and all!
Posted by: Bardiac | September 28, 2006 9:03 PM
I congratulate you PZ, you've outdone yourself here...really.
I have taken this opportunity to nominate you as the nutcase academic of the week..and have full confidence that you will survive to take that coveted yearly prize.
http://restraininorder.blogspot.com/2006/09/for-my-next-trick-ill-set-fire-to.html
Posted by: TJSwift | September 28, 2006 9:17 PM
By the way PZ...care to disclose just how much this breakthough in biological science cost the taxpayers of Minnesota?
Round numbers'd be dandy.
Posted by: TJSwift | September 28, 2006 9:32 PM
In case anyone was wondering, this article is 46% Evil, according to the gematriculator .
A little under half of those zebra fish are going straight to Hell. So the teetotaler is probably ok, and the alcoholic will mostly go to Hell. Except for a little sliver which will go to heaven.
You'll have to plug in the URL yourself to see the detailed analysis.
Posted by: llewelly | September 28, 2006 10:17 PM
you whine a lot, TJ, but do have the slightest clue what you're whining about?
tell us genius, why would a developmental biologist use chemicals to study patterns of development?
do you REALLY think what PZ was doing was simply trying to see if there was a negative effect of alcohol on development? If so, methinks you have spent too much time drinking yourself.
you truly are making us all laugh at you, without realizing it, apparently.
Posted by: Ichthyic | September 28, 2006 10:28 PM
Here is something interesting:
http://www.livescience.com/humanbiology/060927_lazarus_cell.html
Posted by: Kagehi | September 29, 2006 12:05 AM
This "TJ" fellow expects to be taken serious helming a blog entitled "Pair 'o Dice." And yes, that is how he spells it, with the apostrophe before the o.
Posted by: darrell | September 29, 2006 12:32 AM
"And yes, that is how he spells it, with the apostrophe before the o."
Well, I have in my time used many a pair "fo" dice. I wonder if he finds *that* embarrassing.
Posted by: plucky punk | September 29, 2006 1:09 AM
I grimace at the thought of having my "pair" used fo' dice.
And more on topic: The video is absolutely awesome. I second Mr. Carlson's comment...does anyone know of a dump site for sweet bio videos?
Posted by: darrell | September 29, 2006 1:23 AM
I thought alcohol would lead to birth defects. Slow birth doesn't indicate that. Also what are the statistics? One sample does not a Nature paper make. Great post though.
Posted by: Rob | September 29, 2006 2:32 AM
I wonder how it could be possible to stabilize the orientation of the embryo so that the movie wouldn't end as one swirling mess....
Posted by: T_U_T | September 29, 2006 4:59 AM
Carrying on from T_U_T's comment:
Even if it weren't possible to physically stabilise the embryoes, it'd certainly be within the scope of current image processing techniques to automatically reorient each frame and remove that spinning.
Posted by: DrFrank | September 29, 2006 5:11 AM
Yes, alcohol causes serious birth defects. We have lots of data, most of it is statistical: tables of the frequency of classes of problems with given exposure times and concentrations. The movies are just to illustrate some of the phenomena.
The major defect is craniofacial -- branchial arches are malformed. This disrupts circulation, and the heart fills with fluid until it goes boom. We've also got data on branchial cartilage morphology.
These were shot with a fairly low power objective -- 10x -- on a Leica research scope. The embryos are about a millimeter in diameter.
Image processing won't fix the spinning. It's not 2D -- it's the animal flexing and kicking itself around. Sometimes it'll be nose up, sometimes on its left side, sometimes its right.
We do have ways to fix it. We can anesthetize it. For this movie, though, we wanted to compare the onset of activity. Another is to remove the chorion and imbed the embryo directly in agar; we sometimes do that to block the rotation you see early in the video. That constrains the shape of the animal, though, and is only good for shorter recordings.
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September 29, 2006 7:54 AM
Just a question: Isn't 3% a bit overkill? 0.3 would perhaps be more relevant? Oops... that was two.
Posted by: baloo | September 29, 2006 8:04 AM
You'd think so, wouldn't you? 3% is the concentration that most reliably gives us recognizable errors in development. Anything below 1%, the fish, near as we can tell, are perfectly normal -- in fact, they do a little better than the controls (we think the alcohol inhibits fungi and microbial parasites; we don't do it routinely because we don't know that there aren't deleterious effects we can't recognize.)
The fish have amazing powers for processing alcohol. Above 3%, we see significant early mortality, but we've actually had a few survive 6 hours in 7% alcohol. Whatever you do, don't get in a drinking contest with a fish, you won't stand a chance.
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September 29, 2006 8:17 AM
I see... As for fish and alcohol you probably know that the crucian carp is the meanest? When it is deprived of oxygen it switches to glycolysis, but instead of stopping the process at the lactate step, it proceeds to ethanol, which is then secreted over the gills. This happens in winter, and as long as it has enough glycogen, it survives the whole winter without oxygen, but with a constant concentration in the blood of around 10mM. Whether it is more resistant to alcohol during development I have no idea, but it could be interesting to find out...
Posted by: baloo | September 29, 2006 8:52 AM
Wll, t's bvsly wrng t xps fsh mbrys t lchl bcs thy rn't vr prt f thr mthrs' bds. Hmn mbrys R prt f thr mthr's bds, hwvr, nd th mthr hs th rght t d t hr bdy whtvr sh wnts, s dn't knw whr y gt ff syng tht wmn shldn't drnk drng prgnncy (n xcss r thrws). Wht r y - sm nt-chc Chrstn fndy, PZ?
Posted by: Jason | September 29, 2006 9:07 AM
"Where he gets off," Jason, is that it affects fetal development. Can't you read?
Posted by: speedwell | September 29, 2006 9:29 AM
brtn ffcts ftl dvlpmnt, t. s PZ gnst tht, t? f crs nt!
Posted by: Jason | September 29, 2006 9:33 AM
Please do not attempt to engage the trolls in discussion.
Posted by: Caledonian | September 29, 2006 9:44 AM
Ys, pls cntn t ddg nd vd my lgcl, chrnt pnts bt yr lck f cnsstncy.
Posted by: Jason | September 29, 2006 9:58 AM
Image processing won't fix the spinning. It's not 2D -- it's the animal flexing and kicking itself around. Sometimes it'll be nose up, sometimes on its left side, sometimes its right.
Aha, of course - I blame the early morning lack of caffeine for not thinking of that ;)
Posted by: DrFrank | September 29, 2006 10:21 AM
why do i think that Jason's mother drank a lot when she was pregnant with him?
Posted by: Ichthyic | September 29, 2006 10:23 AM
And doesn't that affect the development ? Especially development of the brain ?
Posted by: T_U_T | September 29, 2006 10:28 AM
Please don't say things like that. I take birth defects seriously, and most importantly, I don't condemn blameless individuals born with congenital problems. Jsn had to work hard to earn the contempt he receives.
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September 29, 2006 10:31 AM
Yes, that's another reason we don't use the anesthetic in these studies. We don't know what the long-term effect of the anesthetic might be, and we also don't know about possible interactions between MESAB and ethanol.
In the short term, though, we don't see any obvious effects. Activity is very vigorous during the period 18-24hours, though, and that's also when motor pathways are being set up, so I've long wondered if there wouldn't be deficits in behavior if we suppressed that activity during a critical period. Haven't done the experiments, though...maybe some ambitious student will propose testing it sometime.
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September 29, 2006 10:36 AM
Gives new meaning to "drinks like a fish". These videos are really cool. Thank you so much for posting them. I've never seen anything like it.
Posted by: James Allen | September 29, 2006 12:13 PM
"do you REALLY think what PZ was doing was simply trying to see if there was a negative effect of alcohol on development?"
I'm sorry.
Perhaps I am reading something into "We just wash the embryo with whatever substance we're interested in testing, and see if and how the cells react" that isn't there.
PZ,
It occurs to me that the science of dousing living embryos with alcohol has been pretty well fleshed out, but it appears that brake fluid has been completely overlooked...and charcoal lighter fluid, and drano and sulphuric acid and.....well you get the picture.
Can we be looking forward to some very lively film clips, fresh from the lab?
Posted by: TJSwift | September 29, 2006 12:39 PM
TJ, can you please fix the apostrophe in your blog title? It's bugging my latent OCD.
Posted by: plucky punk | September 29, 2006 12:49 PM
The banner graphic was a gift of time from a friend.
I don't have the tools to fix it myself, and I simply cannot bring myself to cast dispersions on someone who's done me a favor...forget the banner, read the posts and become a better informed individual.
Posted by: TJSwift | September 29, 2006 1:15 PM
Really? Then why are you sitting on your hands? Children are being born every day with fetal alcohol syndrome (FAS), which manifests physically, physiologically, and psychologically in ways from subtle to extreme impairment. You could be helping them, and instead you're just sitting on the sidelines sniping.
Since you have it all figured out, and you're not sharing the secret with all the researchers who are trying very hard to understand all the complex ways in which these factors can interact, I can only conclude that you and Jsn don't want to better understand FAS through studying the complexities of teratogenic effects on systems, because you just hate on FAS kids, and don't want us to find treatments for them.
Or perhaps I misunderstood--you could be making the more sophisticated argument that because of the interspecies differences through which knowledge doesn't translate linearly from the bench to the clinic, that instead of on model organisms, we should be cutting open the FAS kids themselves, in order to better understand the effects.
Which one was your point, again?
Posted by: RavenT | September 29, 2006 1:21 PM
Alright! The first serious attempted explaination.
It's ridiculously specious, but I've got to admit; it's got to to look a lot better on a funding request than " We['re]just wash[ing] the embryo with whatever substance we're interested in testing, and see if and how the cells react".
Posted by: TJSwift | September 29, 2006 1:46 PM
No, I am seriously asking, TJ, because there are currently no good treatments for the deleterious effects of FAS. Since you claim the cellular damage and the corresponding cascades in physiological pathways is so thoroughly understood, please share that understanding with us, to give us something to work with.
If you won't share your understanding with those of us who are working to translate bench findings into clinical treatment, we can only speculate what your reason for withholding it might be. I tend to go with your and Jsn hating on FAS kids, but I could be persuaded that you think that we ought to experiment on FAS kids because the model organisms are insufficiently similar for your thorough understanding.
Or there's a third possibility--you're a libertarian who thinks the FAS kids should just wait for the market to deliver a solution. Come to think of it, I could believe that one, too.
Posted by: RavenT | September 29, 2006 1:58 PM
" We['re]just wash[ing] the embryo with whatever substance we're interested in testing, and see if and how the cells react".
hmm, I'm looking, looking....
i don't see where PZ says that's what he's doing.
can you point out where you mined that quote from in the post?
I really can't find it.
you really don't get it do you?
the idea of testing chemical concentrations on embryos is to see specifically which aspects of development are affected and how.
go figure, we actually learn things from that about how development proceeds, and get more clues about mechanisms.
alcohol isn't a random choice, duh.
you might get a clue as to the specifics of what he was looking for at the end of the post, when PZ says:
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.
you take a simplified presentation as being evidence of an overly simplistic experiment.
bad swifty! bad!
Posted by: Ichthyic | September 29, 2006 2:13 PM
Very cool video. I really enjoyed watching the embryo on the right develop, as it was on its side. I often see the later stages of early development at in my lab (those fish look a lot like our chick embryos!), but it's much better to watch in motion.
Posted by: emily sours | September 29, 2006 2:13 PM
The phrase you're looking for is "cast aspersions." "Dispersion" means "to scatter or disseminate." An aspersion is a derogatory remark. "TJ Swift appears to be an illiterate dimwit with delusions of competency" is an example of an aspersion. I hope you're now clear on the difference.
Posted by: Jason L. | September 29, 2006 2:48 PM
PZ Myers: Do I understand correctly that adult fish also have the incredible ethanol metabolism?
Posted by: Keith Douglas | September 29, 2006 9:19 PM
I don't know! I've read other's work that shows that they are much more tolerant of high blood alcohol levels than we are, but I haven't done that kind of work on our adults. I have a breeding colony only, and I do experiments on the profligate quantities of embryos they produce. I'd have to get IACUC approval to do those kinds of experiments on adult fish.
I also like my fish, and am reluctant to do anything that would harm them, such as exposing them to possibly fatal concentrations of alcohol.
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September 29, 2006 9:38 PM