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Old 21st January 2017   #5 (permalink)
GeorgeAquatics
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Default Re: "Firefly" Cell-transplanted A. mexicanum being made for the pet trade?

Update from Lloyd Strohl (note: I have spoken to him and he says that his response was not directed towards me for writing this article, rather towards the people who were making rude and misleading suggestions in various Facebook groups)

"Lloyd Strohl II added 2 new photos.
November 24, 2016
I offer the following information regarding the infamous “firefly axolotls”.

Please note that I will not reveal details of the complete process involved, any more than I will explain how to make fluorescent human babies (easy, but very expensive), make dinosaurs from emus (very hard and time-consuming, but relatively cheap with the discovery and implementation of CRISPR-Cas9), or destroy every malaria-transmitting Anopheles mosquito on Earth (moderately easy, moderately expensive, and very tempting) because irresponsible people with a little knowledge are dangerous.

First; There are ONLY SIX FIREFLIES. There are three with the GFP tails, and three with normal, non-GFP tails.

Second; Why did I make them?

The short answer is; I am doing a preliminary investigation into the distribution and activation of melanocytes in leucistic axolotls and, in particular, in mosaics. The long answer follows:

I started this investigation after observing several things that seem to contradict accepted wisdom regarding the effect of the leucistic gene in axies, as well as the cause of mosaicism in these animals. The first conundrum resulted when I bred a homozygous melanistic wild-type animal to a golden albino, which should invariably produce only black/spotted offspring. Two of the offspring were not black, but appeared to be leucistic. As these two grew, broad patches of dark pigment became obvious, and I realized they were actually mosaic. The problem with this is; If the conventional wisdom that mosaicism is due to the fusion of two eggs to form one individual (as has been proven conclusively to occur in many organisms, including humans), then these beautiful little mosaics are impossible. Given that the leucistic gene is recessive to the melanistic or wild-type, there could have been no leucistic eggs to join with the black ones, since the mother did not carry that gene.

I began reading everything I could find regarding the genetics of axolotls. This turned out to be a short and pretty unproductive read, failing to answer any questions regarding mosaicism in axies, because there simply hasn’t been any significant research into this. Dr Malikinski, former head of the IU axolotl colony (and original source of my first two animals) directed me to Dr. Randal Voss, geneticist and director of the colony at the University of Kentucky. Randal was kind enough to provide some information, and suggested that my particular mosaics, at least, may be the result of methylation early in development.

During this time I was fortunate to find several more mosaics among my animals, varying tremendously in appearance and distribution of melanocytes. I noted that melanocytes were clearly visible (under a microscope) in the white patches of these animals, but that the melanocytes failed to grow the branching dendrites typical of these cells in darker patches, and most eventually died or failed to produce any melanin to reveal their presence. Unfortunately, I could not draw any realistic conclusions as to the cause of this, since I could not know exactly what genes had been methylated. For example; were the white patches really leucistic, or albino?

According to common wisdom on the subject, the leucistic axolotls produce melanocytes along the neural crest, as all vertebrates do early in development, but these cells fail to migrate away from the neural crest and spread over the body. If this is true, then how did melanocytes end up in the white patches of my mosaics, and what killed them or shut them off?

One of my animals, in particular, piqued my curiosity. He first caught my eye because the GFP gene is expressed only on his right side – not on the left. By definition, then, he is a mosaic, and what many American axolotl enthusiasts call a “chimera”. He appeared, otherwise, to be a normal wild-type. A he grew, however, small pale patches became noticeable and grew into distinct white spots. Within these spots were melanocytes that were pale and nearly free of dendrites.

I decided that the only way to continue my investigation was to produce mosaic animals synthetically. This way, I could be certain of the genetic components of the cells in each patch of tissue.

I had seen pictures of the black-headed, white-bodied axolotls produced by a team of Japanese researchers back in the ‘90s, and had read research in which axolotl embryos had been fused to produce chimeric animals for a study of a lethal version of the gene that controls myofibrillogenesis.

Using these studies and others on various species of salamanders and frogs as a guide, I set to work to produce chimeras. The university studies had succeeded in producing chimeras, but at very low rates of success. George Malakinski and others cited success rates lower than 2% (ususally much lower), meaning that most fused embryos failed to fuse properly and died or had to be destroyed. While subsidized university studies could invest that sort of time and material, I could not, so I worked with small numbers of embryos, methodically testing different methods, modified ringer’s solutions, and antibiotics at different concentrations until I was confident I could do it reliably.

I will not describe the process I ultimately worked out. It is expensive and time-consuming, but I can now fuse embryos with relative ease and at a very high success rate.

The first one and his dark-tailed leucistic sibling (sharing four parents) were produced as a proof-of-concept for my investigation of the pigment distribution and activation in leucistic axolotls. Actually, the dark-tailed lucy was deliberately produced; The firefly was really just a byproduct of that process. You see, for every firefly there is also a dark-tailed lucy – the actual goal of the embryo fusion. If this investigation were being done in a university or commercial laboratory, the wild-type and melanoid embryos used as the cell donors for this investigation would have been destroyed, since they were not the focus of the investigation. I saw no reason to do that, since I knew that both the “donor” embryo and the recipient could live out their happy, healthy lives with just a little additional effort, so that’s what I did. I am keeping the original two animals (See Pic labelled "Figure A"), which are now about six months old, and the other dark-tailed lucies (See Pic labelled "Figure B") – at least until they’ve matured and fulfilled their purpose.

So, you see, the fireflies are not the goal, but a by-product of my investigations. I have sold them because I am an underpaid, overworked teacher, not a wealthy entrepreneur, and (a) cannot afford to keep them all properly and (b) need cash to buy equipment.

And yes, there will be more produced now and then as I continue my efforts.

One of my students recently told me she had read an article online regarding someone “mass-producing” firefly axolotls for the pet trade. I looked into it, and was a bit startled how far this “discussion” has gone online. The article included pics I had posted on Facebook of my animals. My personal favorite statement was that I had “gone all Breaking Bad”.

Yes, that’s right; My wife is even now piling our ill-gained axolotl cash on pallets in a storage locker because there’s just too much to launder away.

I don’t, on principle, argue with internet Trolls. I realize most of them are (a) incapable of grasping the value of evidence and logical argument, and (b) are simply trying to validate their sad, lonely lives as they sit in their mother’s basement in the cool glow of the six-year-old laptop she gave them for their 40th birthday. On the other hand, I am a teacher and academic at heart, and get very disturbed when I see misinformation spread as truth..."

--- https://www.facebook.com/lloyd.stroh...87705631327797
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