22 May 2012

Never send a sibling to do a clone’s job: Part 1

Today, a guest post from Kyle McLea, who has been carrying the Marmorkrebs banner for this round of #SciFund.



As a crustacean biologist who has chiefly studied crabs and lobsters, I’ve been fascinated by the marbled crayfish (Marmorkrebs) for years and for a number of reasons that I’ll detail below. But the first thing that attracted me was simply this.

Really? A crayfish that clones itself?

When Doctor Zen asked me to guest blog on the Marmorkrebs blog I was sure I had a lot to say about my interest in these fascinating critters, but I also wanted to speak to the wider interest of the scientific community. In short: Why should scientists (and the public) care?

To which I say: Never send a (mere) sibling to do a clone’s job.

Let me explain.

Marmorkrebs are a parthenogenetic crayfish. All known examples are female and reproduce themselves entirely without sex. This makes it the first example of a decapod crustacean (and there ~15,000 known species of crabs, lobsters, shrimp, and crayfish!) that only reproduces asexually. There have been a couple of other examples of crayfish that have been found to be genetically identical or to sometimes reproduce asexually, but marbled crayfish were the first found to procreate this way exclusively.

So, in a word, we’re dealing with clone crayfish.

To anyone who’s been following the molecular biology revolution over the past 30ish years, the word clone has both amazing power and amazing misuse and confusion. Clone can mean a lot of things. In this case, I’m not talking about cloning genes (experimentally extracting pieces of nucleic acid, sequencing them, and using them to assemble recombinant—new—combinations of DNA sequence for introduction to a different organism) or about using somatic-cell nuclear transfer to clone embryos (like Dolly the sheep). What I’m saying is that mother and daughter marbled crayfish should be genetically identical, or nearly so, much like identical twins in humans.

It turns out that scientists have thought a lot about the benefits of studying genetically identical individuals in a population, pretty much for as long as there has been a science of genetics. Case in point for inbred (nearly genetically identical) organisms:

“Just as the purity of the chemical assures the pharmacist of the proper filling of the doctor’s prescription, so the purity of the mouse stock can assure a research scientist of a true and sure experiment...In experimental medicine today... the use of in-bred genetic material... is just as necessary as the use of aseptic and anti-septic precautions in surgery.” —C.C. Little, 1936

Now Little may have held some ill-conceived notions about eugenics and the role of tobacco in causing cancer, but on the importance of genetically-identical (isogenic) and inbred laboratory strains he was a pioneer. His work to build up the Jackson Laboratory in Bar Harbor, Maine (the mecca of 5000 unique strains of mice), was a key part of the rise of defined strains and breeds of laboratory animals that continues today.

Those laboratory strains serve as “models” for various human diseases and for particular functions of human physiology, such as immunity or heart function. Among the model organisms listed at NIH, for instance, everything from yeast to mouse and Daphnia to zebrafish can help us to learn about different aspects of human biology and beyond.

But in each case the normal biological variation within an organism confounds us and complicates our study. Unless we want to see the full extent of biological variance (and sometimes we do—safety and efficacy testing of drugs on different populations, anyone?), having organisms that are as close to identical as possible is (often) the goal.

For mice and rats, you might have to backcross (do parent/offspring matings) for more than 20 generations to have a sufficiently inbred line to call it “genetically identical” (and I suspect some gene variation may still exist). Now, mice breed fast, but 20 generations is still real time (years) in the life of a scientist. In the mean time, scientists resort to use of siblings, littermates, or much less inbred animals. These animals have many more differences at the genetic level.

I know you’re seeing where I’m going with this… if an organism started out genetically identical, you’d have a great edge in using this organism to study any number of interesting things. Bringing us back to Marmorkrebs.

Lines of the marbled crayfish are believed to be genetically identical because they do not participate in the normal exchange and shuffling process that accompanies sexual reproduction. But of course, mutations happen to us all. So they’re not likely to be absolutely 100% identical between individuals, but much closer than anything else in the world except for natural identical twins/triplets/etc.

So, I say, send in the clone crayfish. With these animals, we don’t need to compare results with a genetically-different sibling or with an unrelated animal. This unique animal can not only be a useful biological model, but we also don’t have any easily-reared decapod crustacean that can really compete with marbled crayfish, if you take into account their genetic identity.

While they might not be a great model organism for general human physiology (they are crustaceans, after all), there are some specific ways they can help us understand human physiology (e.g., nerves) and they have a lot of other potential uses as a model organism.


Kyle, scientist at Colorado State University and science blogger at By Way of Science. Come back next week for Part 2!

15 May 2012

How many names do you need?

The question of what to call Marmorkrebs in the scientific literature has bubbled up again. When I started the blog, there was no proper scientific name, and I suggested using the name “Marmorkrebs,” because it was distinctive. I thought the matter was relatively settled, scientifically speaking, when Martin and colleagues proposed Procambarus fallax f. virginalis as a scientific name for Marmorkrebs.

A paper by Johnson and colleagues poses a strange puzzle. For some unknown reason, they coin an entirely new name for Marmorkrebs: Procambarus sp. malgasy. Nobody else has used this terminology, although Jones and colleagues do refer to “Malagasy Procambarus” in their paper on Marmorkrebs in Madagascar. Maybe it was meant to be Procambarus sp. Malgasy, with the “Malgasy” purely as an descriptive adjective. Then someone at the editing or proofing stage changed the formatting to resemble a species name.

Even so, it doesn’t explain why they wouldn’t refer to Marmorkrebs as “P. fallax f. virginalis.” They have clearly read the paper by Martin and colleagues – it’s in the list of references.Plus, the analysis by Johnson and colleagues supports that Marmorkrebs is most closely related to P. fallax.

Reference

Jones JPG, Rasamy JR, Harvey A, Toon A, Oidtmann B, Randrianarison MH, Raminosoa N, Ravoahangimalala OR. 2009. The perfect invader: A parthenogenic crayfish poses a new threat to Madagascar’s freshwater biodiversity. Biological Invasions 11(6): 1475-1482. http://dx.doi.org/10.1007/s10530-008-9334-y

Johnson GT, Elder JF, Jr., Thompson SM, Hightower P, Bechler D. 2011. Phylogeny of the freshwater crayfish subfamily Cambarinae based on 16S rDNA gene analysis. Current Trends in Ecology 2: 97-113. http://www.researchtrends.net/tia/abstract.asp?in=0&vn=2&tid=66&aid=3396

Martin P, Dorn NJ, Kawai T, van der Heiden C, Scholtz G. 2010. The enigmatic Marmorkrebs (marbled crayfish) is the parthenogenetic form of Procambarus fallax (Hagen, 1870). Contributions to Zoology 79(3): 107-118. http://dpc.uba.uva.nl/ctz/vol79/nr03/art03

12 May 2012

SciFunded... again!

A big congratulations to Kyle McLea, whose #SciFund Marmorkrebs research project, Crayfish Clone Wars, is now funded.

More impressive, Kyle not only hit his target early, he kicked it a few times and knocked it over for good measure.

Just because a project has met its target, however, does not mean that you can’t still give a donation. You can continue to fuel #SciFund projects until the end of May!

For more crustacean goodness you can support on #SciFund, consider:

A Climate for Castrators? by Alyssa Gehman, which is about the mud crab, Eurypanopeus depressus, and its parasite, Loxothylacus panopeus.

Beach of the Goliath Crabs by Zen Faulkes (me!), which is about the sand crab, Lepidopa benedicti.

08 May 2012

Johnson and colleagues, 2011

Johnson GT, Elder JF, Jr., Thompson SM, Hightower P, Bechler D. 2011. Phylogeny of the freshwater crayfish subfamily Cambarinae based on 16S rDNA gene analysis. Current Trends in Ecology 2: 97-113. http://www.researchtrends.net/tia/abstract.asp?in=0&vn=2&tid=66&aid=3396

Abstract

Freshwater crayfish have been a mainstay in biological experiments as a model species ever since Huxley’s seminal publication The Crayfish. Crayfish have been used in research ranging from vision pigment studies to neural physiology. Non-native species have been introduced on four continents due to their immense economic value. Although crayfish taxonomy is reasonably well resolved at the highest levels, there are many problems at the levels of genus and species. New exploration, technology and methodology have led to the discovery of not only new species but to a phylogenetic complexity that would not have been imagined in Huxley’s era. This complexity is caused by the conservatism of some morphological characters, high intraspecific diversity and convergence. The ambiguity of crayfish taxonomy is particularly evident for species native to South Georgia and North Florida, which are centers of crayfish diversity. Molecular phylogenetic analyses were employed to provide insight into three aspects of crayfish phylogeny. Using partial data from the 16S ribosomal gene, we determined: (a) the evolutionary relationships of a previously unanalyzed species, Procambarus spiculifer, (b) relationships within the genus Procambarus, and (c) the phylogeny of the entire subfamily Cambarinae. The resulting maximum likelihood tree produced phylogenies that were significantly different from the traditional systematic representation of relationships within the subfamily. Specifically, we show that the subfamily Cambarinae should not be divided into three distinct clades according to the genera Procambarus, Cambarus, and Orconectes. While most members of the genus Procambarus cluster within a single monophyletic clade, the genus Orconectes comprises a parayphyletic grouping that appears to also include members of the genus Cambarus.

Keywords: Procambarus spiculifer • Cambaridae • 16s rDNA • astacidae • paratacidae • phylogenetics

Note: Marmorkrebs is referred to in this paper as “Procambarus sp. malgasy.”

01 May 2012

SciFund 2 begins!

SciFund is back! Hosted once again by the fine folks at RocketHub, there are a whole mess of scientific research projects for you to support!

Look for Kyle McLea’s Marmorkrebs project, Crayfish Clone Wars!

You should go to RocketHub now and support scientific research!

25 April 2012

Martin and Scholtz, 2012

Martin P, Scholtz G. 2012. A case of intersexuality in the parthenogenetic Marmorkrebs (Decapoda: Astacida: Cambaridae). Journal of Crustacean Biology 32(3): 345-350. http://dx.doi.org/10.1163/193724012X629031

Abstract

We describe an intersex specimen of the Marmorkrebs, the only obligate parthenogenetic freshwater crayfish with an all-female population. The individual was a fully functional female which possessed male-like first pleopods. Nevertheless, it reproduced successfully and the offspring were normally developed parthenogenetic females, lacking any trace of male traits. The general rarity of aberrant sexual traits in freshwater crayfishes, in particular in Procambarus, is discussed. We suggest that a dysfunction of the sex determining system, which controls the anlagen of the androgenic glands during development, caused the partial male-like phenotype of this Marmorkrebs specimen. The application of this organism for investigations of sex determination and differentiation is recommended.

Keywords: aberrant secondary sexual characteristics • androgenic gland • gonochorism • gynandromorphy • hermaphroditism

23 April 2012

Soedarini and colleagues, 2012

Soedarini B, Klaver L, Roessink I, Widianarko B, van Straalen NM, van Gestel CAM. 2012. Copper kinetics and internal distribution in the marbled crayfish (Procambarus sp.). Chemosphere 87(4): 333–338. http://dx.doi.org/10.1016/j.chemosphere.2011.12.017

Abstract

Metal pollution e.g. copper, in water bodies occurs worldwide. Although copper is an essential trace metal, at certain levels it is still considered as pollutant. The aim of this study was to investigate the effect of exposure concentration on copper bioaccumulation in marbled crayfish (Procambarus sp.) by determining uptake and elimination kinetics. Crayfish were exposed to sub-lethal copper concentrations (average measured concentrations of 0.031 and 0.38 mg Cu L−1) for 14 d and transferred to copper-free water for another 14 d. At different time points during the uptake and elimination phases copper concentrations were measured in five organs (exoskeleton, gills, muscle, ovaries and hepatopancreas). At 0.031 mg Cu L−1, copper levels in the crayfish organs were not significantly increased compared to the control animals, suggesting effective regulation. Exposure to 0.38 mg Cu L−1 did lead to not significantly increased copper levels in muscles and ovaries, while the gills and exoskeleton, which are in direct contact with the water, showed significantly higher copper concentrations. In these four organs, copper showed fast uptake kinetics with equilibrium reached within 10 d of exposure. Copper accumulation was highest in the hepatopancreas; uptake in this storage organ steadily increased with time and did not reach equilibrium within the 14-d exposure period. Copper accumulation levels in the marbled crayfish found in this study were hepatopancreas > gills > exoskeleton > muscle.

Keywords: copper • toxicokinetics • bioaccumulation • hepatopancreas • marbled crayfish