How the bat got its wing.

by Stephen F. Matheson
Originally posted on Quintessence of Dust, May 2008.

Nothing can be more hopeless than to attempt to explain this similarity of pattern in
members of the same class, by utility or by the doctrine of final causes. The hopelessness
of the attempt has been expressly admitted by Owen in his most interesting work on the
'Nature of Limbs.' On the ordinary view of the independent creation of each being, we can
only say that so it is;—that it has so pleased the Creator to construct each animal and
plant.

The explanation is manifest on the theory of the natural selection of successive slight
modifications,—each modification being profitable in some way to the modified form, but
often affecting by correlation of growth other parts of the organisation. In changes of this
nature, there will be little or no tendency to modify the original pattern, or to transpose
parts. The bones of a limb might be shortened and widened to any extent, and become
gradually enveloped in thick membrane, so as to serve as a fin; or a webbed foot might
have all its bones, or certain bones, lengthened to any extent, and the membrane
connecting them increased to any extent, so as to serve as a wing: yet in all this great
amount of modification there will be no tendency to alter the framework of bones or the
relative connexion of the several parts.

– from On the Origin of Species, 1st Edition (1859), Charles Darwin

The wing of a bat is an amazing thing. It's not just a wing; it's clearly a modified mammalian limb. A
bat looks like a lot like a rodent with really long, webbed fingers on elongated arms.

Image from Animal Diversity Web at the University of Michigan.

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Recent genetic analyses have yielded a fairly solid outline of the evolutionary history of bats, which
have left a somewhat poor fossil record in which the earliest fossil bats look pretty much like modern
bats. It seems that bats arose relatively quickly during evolution, acquiring their distinctive feature –
powered flight – in a few million years. No transitional forms have yet been found, which is a shame,
because this particular evolutionary transition is the kind that is otherwise reasonably approachable
for the detailed study of how changes in form come about.

The fossils can't yet show us how paws gave rise to wings, but that doesn't mean we can't test specific
hypotheses regarding the paths that evolution could have taken. In fact, developmental biologists have
enormous resources that can be brought to bear on the question, by virtue of decades of research on
the development and genetics of the wingless terrestrial bat better known as the mouse. In early 2008,
an interesting report described one kind of genetic change that can lead to bat-like bodies, and the
findings put some new wind in the sails of evo-devo.

Two of the more remarkable aspects of bat wing structure are the forelimbs and the forelimb digits,
what humans would call the arms and the fingers. Both are dramatically elongated in the adult animal,
despite getting off to a very typical start during early development. Check it out: in the picture below,
bat and mouse limbs are compared with the image scaled so that body lengths are comparable.

Image from Figure 1 of Cretekos et al., cited below.

Developmental biologists have some pretty good ideas about how this might arise physiologically:
certain growth factors (called bone morphogenetic proteins, or BMPs) are known to control limb
growth, and some BMPs seem to be turned up in developing bat fingers. But the genetic mechanisms
underlying these processes are unknown.

Enter Chris Cretekos and colleagues, then working in a group in Houston headed by Richard
Behringer. They set out to examine the genetic underpinnings of the elongation of the forelimbs
(arms) of bats, using the formidable tools of mouse developmental genetics. And, clearly, they also
sought to directly test one of the central hypotheses of evo-devo: that changes in regulatory DNA
sequences (as opposed to changes within the genes themselves) are a potent source of variation in
evolution. Consider the beginning of their abstract:

Natural selection acts on variation within populations, resulting in modified organ

morphology, physiology, and ultimately the formation of new species. Although variation
in orthologous proteins can contribute to these modifications, differences in DNA
sequences regulating gene expression may be a primary source of variation.

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 – From C.J. Cretekos et al., "Regulatory divergence modifies limb length between
mammals, Genes & Development 22:141-151, 15 Jan. 2008
Besides their expertise in mouse genetics, the authors brought two major assets to their study: 1) they
had already carefully mapped the development of the short-tailed fruit bat (Carollia perspicillata,
"our model Chiropteran"); and 2) they knew a lot about the genetic control of limb length in other
mammals. In particular, they knew that the protein Prx1 is known to influence limb elongation, by
controlling the expression of other genes. So they hypothesized that changes in the activity or level of
Prx1 might underlie the difference in limb length between bats and mice, and they were well-equipped
to do the experiments.

First, the authors examined the Prx1 gene in the two species, and found that the overall structure of
the gene is very similar in both mice and bats, and that the actual coding sequences of the two genes
are almost completely identical. (Aligning the coding sequences showed that more than 99% of the
amino acids are the same in both species.) In other words, the part of the Prx1 gene that codes for
protein is almost certainly not a source of variation between mice and bats. This could mean that Prx1
doesn't have anything to do with the difference between forelimb length in these two species, or it
could mean the the difference is generated, at least in part, by variation in the regulation of the gene.
Cretekos et al. postulated that altered Prx1 regulation might be involved, and designed a cool
experiment to address this possibility.

They already knew that the Prx1 gene in mice contains known regulatory elements in particular
locations within the gene. (Such elements are often located in the DNA sequences that precede the
coding region.) When they looked at the bat gene, they found similar elements in the same location,
but these elements showed some intriguing variation: when the two regions were aligned, they shared
only 67% identity, meaning that a third of the DNA bases were different in mouse and bat. They did
some nifty cell biology to show that this region did function as a regulator of the expression of Prx1,
then did something that biologists could only dream about before the genomic era: they altered the
mouse genome by replacing the mouse regulatory region with the corresponding region from the bat
genome. In other words, they gave a mouse a piece of a bat's genome, without actually changing the
coding sequence of any gene.

The result was dramatic, although it won't sound that way at first. The mice with the bat DNA
displayed forelimbs that were 6% longer than normal. Why is this a dramatic result? Well, first of all,
think about a 6% change in a major structural attribute. If adult males in a certain country average
5'10" in height, a 6% increase would mean an increase of more than 4 inches. But more importantly,
the Prx1 gene is known to account for about 12% of forelimb length – mice that lack the gene
altogether show a 12% reduction in forelimb length. That 6% change reflects a huge change in Prx1
activity, a change that was completely due to alterations in regulatory DNA sequences without any
change in coding sequence.

If that's not impressive enough, the authors went on to examine the importance of this regulatory
region in mice, by deleting it altogether. The result was very surprising, but very interesting: limb
length in mice was completely unaffected by the loss of this chunk of regulatory DNA. (The region
we're discussing is 1000 bases in length.) This means that the Prx1 gene of both bats and mice
contains a regulatory region that is completely dispensable for normal development but that can be
altered to generate significant changes in limb length, which points to significant evolutionary
potential in genetic regions that seem unimportant. Here's how the authors say it:

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 Maintenance of redundant enhancers for essential developmental control genes would
allow changes in expression pattern to arise from mutations that alter regulatory activity
while preserving the required gene function.

So, why is this significant? Here are two aspects of the story that are worth highlighting.

1. The results provide strong (and rare) experimental support for the ideas of the evo-
devo school. The currently-heated debate over the merits of evo-devo is focused on the central evo-
devo claim that morphological evolution (i.e., evolutionary changes in form) is driven to a large extent
by changes in the regulation of gene expression, and less so by changes in the structures of the
proteins that are encoded. To simplify, evo-devo postulates that significant evolutionary change – like
that discussed here – is more likely a result of the varied use of a protein toolkit than a result of
modification of the toolkit itself. Cretekos et al. have presented a case in point, and one that is
considered outstanding in that it documents a morphological gain; many previous examples showed
only losses.

2. The results provide a sharp picture of what Darwin's vision of "successive slight
modifications" means in terms of developmental biology. In this case, the modifications (of a
redundant regulatory region) can yield significant anatomical remodeling without altering protein
structure at all.

The article was a notable advance for evo-devo and for evolutionary science, but soon there will surely
be many others like it. Desperate or ignorant creationists will always find a way to avoid facing the
explanatory power of common descent, but scientists are just plugging away, and for every blog post
by a creationist ignoramus, there are 30 unheralded publications in the biological literature that
advance our understanding of common descent and the mechanisms that generate biological novelty.
And they're fun to read.

Article(s) discussed in this post:

Cretekos, C.J. et al. (2008) Regulatory divergence modifies limb length between mammals.
Genes & Development 22:141-151.