The Glowing Axolotl

g1g5

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Whilst studying for my undergraduate degree, I read an article in Developmental Biology which gave me more than a little bit of a laugh.

http://www.ambystoma.org/AGSC/article.pdf

It essentially outlines the production of GFP expressing strain of Axolotl.

Aequorea victoria Green fluorescence protein GFP is a biological product with a fluorescence emission peak of 509nm (green), resulting from an excitation peak of 395nm (blue). It is a common analytical tool widely used as expression label in molecular biology.

The concept of GFP expressing advanced macroorganisms is not a new one, GFP mice has existed since 1998 [1]. However the thought of a glowing axolotl must bring a smile to any amphibian hobbyist, and has been highly significant in the study of cell fate in developmental biology. A factor unique to the axolotl is its unique regenerative capability, being able to regrow entire lost limbs, an ability it retains into adulthood [2]. This axolotl tm is therefore of some interest in the study of tissue generation.

Anyway enjoy.

[1]
A.-K. Hadjantonakis, M. Gertsenstein, M. Ikawa, M. Okabe, and A. Nagy. Generating green fluorescent mice by germline transmission of green fluorescent ES cells. Mechanism of Development 76, 79-90 (1998).

[2]
I suspect this is linked to Axolotl's neoteny trait. Does anyone know if adult american tiger salamander could regrow limbs?
 

g1g5

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I accept that there are numerous threads on this topic. However most of them ( at least those I have managed to find) are fairly informal and do not really discuss the scientific merits or implications of this paper, beyond generally saying how cool a pet the glowing axolotl would make (whilst indubitably true this is not an academic analysis).

Another interesting point to highlight is how elegantly simple the protocol is. Most of the method used in this paper is standard microbiological techniques and not beyond the skill of a good undergraduate biotech student. The plasmid vector is fairly common (addgene would probably supply it) and sequence of the GFP containing construct, including the associated regulation regions (I believe they used cytoskeleton actin promoter), can be found in any respectable gene bank. Barring a few basic lab apparatus, the only piece of material needed is Axolotl embryonic cells, which is probably less difficult to get one's hands on than an official lab clearance for vertebrate tissue culture (in the UK at least).

Honestly, for anyone with a reasonable background in molecular bioscience and a supply of embryonic axolotls, and the British Government's bureaucratic cr@p not withstanding, following in this paper's footsteps only requires the will to do so.
 

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Another interesting point to highlight is how elegantly simple the protocol is. Most of the method used in this paper is standard microbiological techniques and not beyond the skill of a good undergraduate biotech student. The plasmid vector is fairly common (addgene would probably supply it) and sequence of the GFP containing construct, including the associated regulation regions (I believe they used cytoskeleton actin promoter), can be found in any respectable gene bank. Barring a few basic lab apparatus, the only piece of material needed is Axolotl embryonic cells, which is probably less difficult to get one's hands on than an official lab clearance for vertebrate tissue culture (in the UK at least).

Honestly, for anyone with a reasonable background in molecular bioscience and a supply of embryonic axolotls, and the British Government's bureaucratic cr@p not withstanding, following in this paper's footsteps only requires the will to do so.

Old topic, I know. But from my experience, the only reason why some molecular biology/biotech methods appear "easy" is because most undergraduate students are provided with fail-proof experiments. Not only that, most scientific articles do ramble on about the time they spent optimising the technique. Once again, this is just based on previous experience.

- Jay
 

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Putting GFP into organisms became something of a fad/"me too" thing before they got to axolotls. It has its uses but I don't think the procedure in and of itself sparked much interest simply because it's nothing really new. I'm a little surprised it even got into Developmental Biology rather than a lower tier journal.
 
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