Signal ‘remodels’ the body from larva to fly

WASHINGTON U.-ST. LOUIS (US) — New research helps explain how biochemical signals prompt the dramatic body changes in a fruit fly’s lifecycle.

Fruit flies go through three main life phases: the larva, the pupa, and the adult. Earlier work had shown that the larval and adult forms are patterned by the same “signaling systems,” or chains of biochemicals that transfer a signal from receptors on the surface of cells to target genes within cell nuclei.

Genes whose expression is induced by pulses of a steroid hormone are key to fruit fly metamorphosis. In this adult male fly, one of these steroid-sensitive genes, E93, is expressed in every tissue (clockwise from top): mouth parts, eye, thorax, abdomen, leg, wing, and antenna. (Credit: Ian and Dianne Duncan)

What scientists didn’t understand was how the same signaling systems could orchestrate the formation of a larva in one case and the adult fly in the other.

In the latest issue of Proceedings of the National Academy of Sciences, Ian and Dianne Duncan, biologists at Washington University in St. Louis show that a gene expressed only in the pupal stage redirects signaling systems so that they activate a different set of target genes than in earlier stages.

This gene is itself controlled by a steroid hormone that turns on many other genes as well. So, insect metamorphosis, triggered by a hormone, resembles puberty, the human analog of metamorphosis, which is also triggered by hormones.

A wholesale change

In 2011, scientists at the University of Cambridge published a paper in the journal Proceedings of the National Academy of Sciences (PNAS).  that described what happened when they artificially turned on a regulatory gene at different stages of a fly’s metamorphosis.

Using microarrays that detected the products of gene activity, they found that at every stage of metamorphosis, the regulatory gene subtly increased or decreased the expression of hundreds of downstream genes. But the kicker was that at different stages of metamorphosis, different downstream genes were turned on.

“They found 870-some target genes,” Ian Duncan says, “and among those 870, roughly 200 were induced in the larva, more than 400 were induced in the prepupa, and 350 were induced in the pupa, but the thing is, the genes controlled at each stage were almost completely different. So they realized there were global changes of rules from one stage to the next.”

“It’s as if two teams were playing soccer,” Dianne Duncan says, “and at halftime the referee comes out and hands out a new set of rules. Now you’ve got the same players, the same field, the same goals, but the teams are playing hockey not soccer. The rules are different, so the game is different.”

The Cambridge researchers ended their article in PNAS by saying that more research was needed to understand how this repurposing of signaling pathways happens. The Duncans, in reply, are publishing their study in the same journal.

Throwing two switches

The Duncans focused on a gene called E93 that is turned on by steroid pulses, but only at the pupal stage. “It is required for all patterning and production of new structures in the pupa, but it doesn’t play any role in making the larva,” Ian Duncan says.

To understand in detail what E93 was doing in the fly, the Duncans chose a simple and well understood patterning process: the activation of a target gene called Distal-less that makes dark spots near the fly’s leg bristles called bracts.

The target gene is activated by a well-studied signaling pathway—the epidermal growth factor receptor (EGFR) pathway. “The EGFR signal pathway is used all over the place in the fly,” Ian Duncan says. “It’s used for different things at different times.”

To demonstrate that E93 had to be activated before the EGFR pathway could turn on the target gene, the Duncans looked at flies with mutated E93 genes.

“In the mutant that doesn’t have this gene, cells don’t respond to the signaling pathway, and bracts fail to form,” Ian Duncan says.

Further experiments showed that E93 and EGFR signaling are both needed to turn on the target gene Distal-less. E93 tells Distal-less when to turn on and EGFR signaling tells it where to turn on. Having to throw two switches ensures that the target gene is activated at only the right time and place.

Turning on bracts at the wrong time

As the “two switch” idea suggests, it isn’t possible to activate Distal-less at the wrong stage by artificially turning on either E93 or EGFR signaling. Only turning on both E93 and EGFR signaling activates Distal-less.

“If both E93 and EGFR signaling are active, the Distal-less gene will turn on in the larva even though it’s the wrong time,” Ian Duncan says.

“This finding was important because it showed that E93 can make cells behave as though they’re in a pupa even if they’re not. E93 says it’s the right time.”

Bracts and brains

Bracts aren’t very sexy, and in fact, the Duncans say, nobody knows what they do. They’re just a convenient anatomical feature to study. They plan next to study the effect of E93 on the fruit fly brain.

“We know enough now to know that E93 strongly affects brain remodeling in the pupa. Presumably E93 is doing the same thing in the nervous system that it is doing in the leg: it’s affecting the responsiveness of genes,” Ian Duncan says.

And there is again a tantalizing parallel to human physiology. Not only are the frontal lobes remodeled during puberty, that’s also when diseases such as bipolar disorder and schizophrenia tend to manifest.

“There’s so much cell death and rewiring during this period,” Dianne Duncan says, “it’s astonishing that we get through it as well as we do.”

“There’s even a human homolog of E93 called LCOR (ligand-dependent co-repressor) that is also involved in steroid signaling,” she says. Scientists don’t know yet what LCOR does biologically, but future research may address its function.

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