Friday Cephalopod: The Ecstasy Protocol

The story of this experiment where octopuses were given a hit of Ecstasy (MDMA) is all over the interwebs right now, but not very many people have bothered to read the original paper, given the weird twist it’s been given, that these bored scientists were giving their pets drugs to see what they’d do. That isn’t the case at all. The first part of the paper is all about sequence analysis of the serotonin transporter gene SLC6A4 (the gene that is affected by MDMA), and a comparison of the gene in different phyla. This research starts with some serious, detailed work on the functional mechanics of the gene. If you want to study the role of serotonin in mood and behavior in humans, comparative work of this sort is essential if you want to puzzle out which bits and pieces of the gene are important.

The starting point of this work is the phylogeny, which tells us that serotonin is an ancient transmitter, and that many of the elements of its signaling process are evolutionarily conserved. It should also tell you that its history is complicated, because there are a lot of duplications and subtle variants.

(A and B) Maximum-likelihood trees of SLC6A transporters (A) and SLC6A4 serotonin transporters (B) in select taxa. Species are mapped to tree and protein identifiers in Table S3. For a larger version of (A), see Figure S1.
(A) A maximum-likelihood “best tree” for the SLC6A gene family. The maximum-likelihood tree produced by RAxML includes 503 proteins and 21 species, with tree building based on a MAFFT alignment of full-length sequences.
(B) The SLC6A4 gene family, a subtree of the maximum-likelihood “best tree” in (A).

Thus, monoamine transporters may represent an ancient innovation that arose early in bilaterian evolution, with various ancient and more recent duplications in different lineages.

We know, though, from the molecular work that the octopus has an SLC6A4 gene, and further, that the portion of the gene that binds to MDMA has been conserved. The next question, then, is to ask how this gene modulates octopus behavior. That’s when the test of exposing them to MDMA was proposed.

Of course, you don’t just give animals the drug and watch to see what happens. You’ve got to have a hypothesis. In this case, prior observations, much of it informal, in a different model system, Homo sapiens, was used to infer that MDMA exposure might increase social behavior (“I love you, man”), so they designed an experimental setup to directly test that behavior. Here it is.

(A and B) Diagrams illustrating timeline (A) and experimental protocol (B) for three-chambered social approach assay.
(C) Quantification of time spent in each chamber during 30-min test sessions (n = 4; two-way repeated-measures ANOVA: p = 0.0157; post hoc unpaired t test pre, social versus center p = 0.4301, object versus center p = 0.0175; post, social versus center p = 0.0190, object versus center p = 0.1781).
(D–K) Comparisons between pre- versus post-MDMA-treatment conditions (paired t test pre versus post, social time p = 0.0274; object time p = 0.1139; center time p = 0.7658; transitions p = 0.3993).
(L) Photograph of social interaction under the saline (pre) condition.
(M) Photograph of social interaction under the MDMA (post) condition.
Error bars represent the SEM.

The design is straightforward: since you can’t ask an octopus to explain the sensations they feel under MDMA, you give them a choice. They have a 3-chambered box, and they put the octopus they were testing in the center box. On the left, there is an object, a toy they can explore. On the right, there is another octopus, an opportunity to socialize. Will they prefer an inanimate object they can tinker with, or to approach a conspecific?

In observations without any drugs, they determined that, as expected, octopuses are relatively solitary animals — in part because nobody likes the males. Both male and female subjects spent most of their time in the central chamber, but would spend more time in the social chamber if the octopus there was a female, but if it was male, they were suddenly much more interested in hanging out with the object.

Nevertheless, somewhat surprisingly, both male and female subjects did exhibit social approach to a novel female conspecific, a finding that may reflect an adaptation of laboratory raised animals or an incomplete ethological description of the full repertoire of social behaviors in the wild. Although we cannot rule out the possibility that the female versus male social object preference effect is governed by relative size differences between subject and social objects, we think this is unlikely since we observed aversion to a male social object both when the subject was greater and smaller in size.

I’m just going to assume that the male aversion was because males are assholes, which is apparently a phylogenetically ancient trait.

What they clearly saw, though, was that under the influence of MDMA, the subject octopus switched to expressing a far greater interest in exploring the social object, whether it was a male or female. The animals responded to the entactogenic properties of the drug, an interesting observation that suggests that this is also a phylogenetically ancient trait.

A word of caution, in interpreting these data: there has been a tendency lately to cherry-pick examples of complex animal behavior to justify specific human social structures. What this work tells us is that there are conserved biochemical pathways that are regulated to trigger behaviors along a continuum — that diverse animals use serotinergic pathways as a kind of slider control, that can be ramped up to increase cooperative, social behavior, or tuned down to increase aggressive, asocial behavior. That this kind of neural regulative control exists, is conserved, and has deep roots in animal evolution cannot be used to argue that humans are naturally supposed to build capitalist dominance hierarchies any more than it can be used to claim that humans are adapted to live in cooperative communes full of peace, love, and understanding. The pathway is present in animals with diverse behavioral patterns.

Monoamine transporters, including human SERT, DAT, and NET, appear to be a bilaterian innovation, suggesting a possible ancient evolutionary role in nervous system centralization and elaboration, both hallmarks of the Bilateria, and the families have undergone complex patterns of gene duplication and loss throughout the clade over time. Phylogenetic analysis revealed clear orthologs of human SLC6A4 in octopuses, as well as high levels of conservation in the transmembrane domain and amino acid region critical to MDMA binding. Interestingly, we found that SLC6A4 is broadly conserved in the fruit fly, the worm, and most other bilaterian animals but is surprisingly absent in both of the eusocial hymenopteran insects, the honeybee and leaf cutter ant.

I’m sure that warning won’t stop everyone, though. I also expect that there will be some humans using it to argue that we all ought to be taking more E, a position that the research does not endorse at all, either.


One last thing I want to mention is a bit from the methods section (yeah, I read the methods): cephalopods are completely exempt from the ethical regulations for the care of laboratory animals, but the investigators followed them anyway.

Care of invertebrates, like O. bimaculoides, does not fall under United States Animal Welfare Act regulation, and is omitted from the PHS-NIH “Guide for the Care and Use of Laboratory Animals.” Thus, an Institutional Animal Care and Use Committee, a Committee on Ethics for Animal Experiments, or other granting authority does not formally review and approve experimental procedures on and care of invertebrate species, like O. bimaculoides, at the Marine Biological Laboratory. However, in accordance with Marine Biological Laboratory Institutional Animal Care and Use Committee guidelines for invertebrates, our care and use of O. bimaculoides at the Marine Biological Laboratory and at Johns Hopkins University generally followed tenets prescribed by the Animal Welfare Act, including the three ‘Rs’ (refining, replacing, and reducing unnecessary animal research).

You can’t punch a kitten with a meat hammer, but you can do it to an octopus, if you want, which seems backwards to me. No! I mean, you shouldn’t be allowed to punch any animal with a meat hammer in the name of science. But once again, our laws are inconsistent and arbitrary.

Squids from SPAAAAAAAAACE!

I was reading this article with a provocative title: Cause of Cambrian Explosion – Terrestrial or Cosmic?. It set my alarm bells ringing from the title onward.

Look at those authors! So many, yet the paper itself is so empty of data. Most I don’t know. Steele I’ve heard of — he was promoting neo-Lamarckism in the 1980s, and thinks the Cambrian explosion was caused by retroviruses squirting new complex genes into the ancestors of all animals. Brig Klyce I’ve bumped into a few times on the internet…he’s a panspermia fanatic. Milton Wainwright is the guy who used an EM to look for odd blobs and declared they are evidence of alien life. The Wallis’s were part of a time that announced that diatoms came from outer space. Oh, and Chandra Wickramasinghe…yes, we have crossed paths multiple times. He published a lot in the Journal of Cosmology, with an editor, Rhawn Joseph, who really, really doesn’t like me.

Wickramasinghe has been making bank on this nonsensical idea that genes for complex intelligent life have periodically rained down on the Earth from outer space. There is no evidence for it, and no reason to invoke this random phenomenon to explain biology — we have random phenomena enough, thank you very much, and none of them have the extreme weirdness of the space virus explanation.

I guess I have heard of quite a few of the authors! And it’s a most unsavory stew of notorious crackpots.

Let’s take a look at the abstract for this gem of a paper, shall we?

We review the salient evidence consistent with or predicted by the Hoyle-Wickramasinghe (H-W) thesis of Cometary (Cosmic) Biology. Much of this physical and biological evidence is multifactorial. One particular focus are the recent studies which date the emergence of the complex retroviruses of vertebrate lines at or just before the Cambrian Explosion of ~500 Ma. Such viruses are known to be plausibly associated with major evolutionary genomic processes. We believe this coincidence is not fortuitous but is consistent with a key prediction of H-W theory whereby major extinction-diversification evolutionary boundaries coincide with virus-bearing cometary-bolide bombardment events. A second focus is the remarkable evolution of intelligent complexity (Cephalopods) culminating in the emergence of the Octopus. A third focus concerns the micro-organism fossil evidence contained within meteorites as well as the detection in the upper atmosphere of apparent incoming life-bearing particles from space. In our view the totality of the multifactorial data and critical analyses assembled by Fred Hoyle, Chandra Wickramasinghe and their many colleagues since the 1960s leads to a very plausible conclusion — life may have been seeded here on Earth by life-bearing comets as soon as conditions on Earth allowed it to flourish (about or just before 4.1 Billion years ago); and living organisms such as space-resistant and space-hardy bacteria, viruses, more complex eukaryotic cells, fertilised ova and seeds have been continuously delivered ever since to Earth so being one important driver of further terrestrial evolution which has resulted in considerable genetic diversity and which has led to the emergence of mankind.

It’s a moderately long paper, because it’s really easy to layer on thick coats of bullshit when you don’t care about the quality of the evidence. So I’m just going to look at — can you guess? — his second focus, “the remarkable evolution of intelligent complexity (Cephalopods) culminating in the emergence of the Octopus”.

It’s garbage.

There are novelties in cephalopod evolution, and I’ve written about them before. In particular, cephalopods carry out a significant amount of gene editing, that is, they use enzymes to modify a few of the bases in RNA before it is translated into protein. This is not a shocking surprise — it’s not a universal modification of every RNA, but it has been observed in phyla all across the animal kingdom — although some gullible sources claim it is a violation of the central dogma (they’re wrong). But the key thing is that it’s not unique to cephalopods, lots of organisms have the enzymes, so you can’t use it as evidence for the claim that gene editing came from outer space.

In particular, there is no reasonable justification for this claim:

Thus the possibility that cryopreserved Squid and/or Octopus eggs, arrived in icy bolides several hundred million years ago should not be discounted (below) as that would be a parsimonious cosmic explanation for the Octopus’ sudden emergence on Earth ca. 270 million years ago. Indeed this principle applies to the sudden appearance in the fossil record of pretty well all major life forms, covered in the prescient concept of “punctuated equilibrium” by Eldridge and Gould advanced in the early 1970s (1972, 1977); and see the conceptual cartoon of Fig. 6. Therefore, similar living features like this “as if the genes were derived from some type of pre-existence” (Hoyle and Wickramasinghe, 1981) apply to many other biological ensembles when closely examined. One little known yet cogent example is the response and resistance of the eye structures of the Drosophila fruit fly to normally lethally damaging UV radiation at 2537 Å, given that this wavelength does not penetrate the ozone layer and is thus not evident as a Darwinian selective factor at the surface of the Earth (Lutz and Grisewood, 1934) and see Hoyle and Wickramasinghe (1981, p.12e13). Many of these “unearthly” properties of organisms can be plausibly explained if we admit the enlarged cosmic biosphere that is indicated by modern astronomical research e discoveries of exoplanets already discussed. The average distance between habitable planets in our galaxy now to be reckoned in light years e typically 5 light years (Wickramasinghe et al., 2012). Virion/gene exchanges thus appear to be inevitable over such short cosmic distances. The many features of biology that are not optimised to local conditions on the Earth may be readily understood in this wider perspective.

We’ve gone from a few viral genes raining down on Earth and getting incorporated into life, to frozen squid eggs drifting from Alpha Centauri to Earth in icy meteors and somehow crashing into our oceans and surviving to populate the seas. I don’t think the authors understand the word “parsimonious”. If this were true, cephalopods would represent an entirely novel lineage, and more than having a few molecular novelties, they would be completely unrelated to any other animal lineage on the planet. They would not be related to other molluscs. They would not be protostomes. They would not be eukaryotes. They would be totally alien.

The authors even seem to be superficially conscious of this problem. Here is the “conceptual cartoon of Fig. 6”.

This diagram is what you get when you pretend that lineages are made solely of apomorphies, or the derived traits that distinguish each species from other organisms, and close your eyes to the plesiomorphies, or shared similarities. A phylogenetic tree is not “forced”, it is produced by identifying shared traits. The octopus has molecular similarities to snails, and the two together have similarities to other invertebrates, and all of them have shared attributes with all animals. You don’t get to just ignore all that! This is equivalent to saying that octopuses have tentacles, therefore octopuses are from outer space, completely neglecting the fact that octopuses have homologous genes linking them to insects and sea cucumbers and people.

To back up the remarkable assertion that cephalopods fell from space, they present no evidence, other than a flurry of citations of … N. Chandra Wickramasinghe. It’s an embarrassingly masturbatory display. Wickramasinghe and his associates have been churning out these useless, garbage papers for decades, and now they use the volume of shit he has produced as evidence that his shit is valid. He occasionally sprinkles in references to other authors, which he gets wrong: Stephen J. Gould would not recognize figure 6 as an accurate representation of punctuated equilibrium. This is not how science is supposed to work. It’s simply fraudulent.

Wickramasinghe used to be associated with Cardiff University — they fired him and closed his astrobiology ‘department’, which turns out to have been a bit of a Potemkin village anyway. It was run entirely by Wickramasinghe as a part-time employee, and the entirety of the staff were “honorary”, unpaid volunteers.

“It was only costing them between £14,000 and £15,000 (about $24,000) a year to retain me as a part time director of the centre.

“All the other staff, totaling about 12, is honorary research fellows and associates who were not costing the university anything at all. They have brought a huge amount of credit to Cardiff University and so it amazed me that the university would discontinue their support for astrobiology. “What they did to me is a travesty of normal university practice and I still don’t understand the motive. I can’t believe for a moment that they are strapped for £15,000 a year to maintain a centre that has, for good or bad, a very high profile internationally. “We continue to make headlines in various things that we do. Some of our work remains controversial but it is in the nature of science to promote controversy as long as it is intelligent controversy. That’s within the rules of the game. If people agree 100 per cent what they’re doing then science becomes a bit insubstantial. “I just fail to understand why they do this. It could be ageism because, at 71, I’m over the retirement age by a couple of years, but I’ve been around for years and have published many papers. I was Sir Fred Hoyle’s longest-running collaborator from the time I was a student at Cambridge.”

Cardiff claims the closure was entirely due to budgetary reasons, but I rather suspect that, contrary to Wickramasinghe’s claim, his slack work and low standards of evidence have frequently brought discredit to the university.

Don’t cry for Chandra, though. He was snapped up by the University of Buckingham to form a “centre for astrobiology”. I think that might mean he was allowed to host a webpage on their site, because he’s never had a real research unit, and I doubt that he’s been given the funds for one now.

But yeah, if you see his name on anything, or apparently any of the names in that long list of articles, you’ve found a treasure trove of pseudoscience.