a-new-approach-that-eliminates-handling-for-studying-aggression-loser

A New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in <em>Drosophila melanogaster</em>

Aggressive behavior in Drosophila melanogaster is composed of the sequential expression of stereotypical behavioral patterns (for analysis see 1). This ...

Harvard Medical School

The myelodysplastic syndromes and myeloproliferative disorders are associated with deregulated production of myeloid cells. The mechanisms underlying these disorders are not well defined.

Mutation in TET2 in myeloid cancers.

The steroid hormone ecdysone influences Drosophila lifespan. Longevity is extended in mutants deficient for ecdysone synthesis or mutants of the ecdysone receptor (EcR). However, the underlying mechanisms remain unclear. Here we conditionally inactivated EcR by RNA interference or expression of dominant negative forms, using the RU486 inducible system. A mild ubiquitous inactivation of EcR during adulthood was sufficient to slow the aging of male flies, whereas a stronger EcR inactivation decreased longevity. Surprisingly, ubiquitous inactivation of EcR strongly decreased female lifespan. This deleterious effect was suppressed in sterile ovo(D1) mutant females, suggesting that EcR represses a negative signal for lifespan produced in ovaries. These results reveal a complex adult and sex-specific control of lifespan by steroid signalling in Drosophila.

The steroid hormone receptor EcR finely modulates Drosophila lifespan during adulthood in a sex-specific manner.

Aversive olfactory memory is formed in the mushroom bodies in Drosophila melanogaster. Memory retrieval requires mushroom body output, but the manner in which a memory trace in the mushroom body drives conditioned avoidance of a learned odor remains unknown. To identify neurons that are involved in olfactory memory retrieval, we performed an anatomical and functional screen of defined sets of mushroom body output neurons. We found that MB-V2 neurons were essential for retrieval of both short- and long-lasting memory, but not for memory formation or memory consolidation. MB-V2 neurons are cholinergic efferent neurons that project from the mushroom body vertical lobes to the middle superiormedial protocerebrum and the lateral horn. Notably, the odor response of MB-V2 neurons was modified after conditioning. As the lateral horn has been implicated in innate responses to repellent odorants, we propose that MB-V2 neurons recruit the olfactory pathway involved in innate odor avoidance during memory retrieval.

Mushroom body efferent neurons responsible for aversive olfactory memory retrieval in Drosophila.

It is broadly accepted that long-term memory (LTM) is formed sequentially after learning and short-term memory (STM) formation, but the nature of the relationship between early and late memory traces remains heavily debated [1-5]. To shed light on this issue, we used an olfactory appetitive conditioning in Drosophila, wherein starved flies learned to associate an odor with the presence of sugar [6]. We took advantage of the fact that both STM and LTM are generated after a unique conditioning cycle [7, 8] to demonstrate that appetitive LTM is able to form independently of STM. More specifically, we show that (1) STM retrieval involves output from γ neurons of the mushroom body (MB), i.e., the olfactory memory center [9, 10], whereas LTM retrieval involves output from αβ MB neurons; (2) STM information is not transferred from γ neurons to αβ neurons for LTM formation; and (3) the adenylyl cyclase RUT, which is thought to operate as a coincidence detector between the olfactory stimulus and the sugar stimulus [11-14], is required independently in γ neurons to form appetitive STM and in αβ neurons to form LTM. Taken together, these results demonstrate that appetitive short- and long-term memories are formed and processed in parallel.

Parallel processing of appetitive short- and long-term memories in Drosophila.

A fundamental duty of any efficient memory system is to prevent long-lasting storage of poorly relevant information. However, little is known about dedicated mechanisms that appropriately trigger production of long-term memory (LTM). We examined the role of Drosophila dopaminergic neurons in the control of LTM formation and found that they act as a switch between two exclusive consolidation pathways leading to LTM or anesthesia-resistant memory (ARM). Blockade, after aversive olfactory conditioning, of three pairs of dopaminergic neurons projecting on mushroom bodies, the olfactory memory center, enhanced ARM, whereas their overactivation conversely impaired ARM. Notably, blockade of these neurons during the intertrial intervals of a spaced training precluded LTM formation. Two pairs of these dopaminergic neurons displayed sustained calcium oscillations in naive flies. Oscillations were weakened by ARM-inducing massed training and were enhanced during LTM formation. Our results indicate that oscillations of two pairs of dopaminergic neurons control ARM levels and gate LTM.

Slow oscillations in two pairs of dopaminergic neurons gate long-term memory formation in Drosophila.

The remodelling of neurons during their development is considered necessary for their normal function. One fundamental mechanism involved in this remodelling process in both vertebrates and invertebrates is axon pruning. A well-documented case of such neuronal remodelling is the developmental axon pruning of mushroom body γ neurons that occurs during metamorphosis in Drosophila. The γ neurons undergo pruning of larval-specific dendrites and axons at metamorphosis, followed by their regrowth as adult-specific dendrites and axons. We recently revealed a molecular cascade required for this pruning. The nuclear receptor ftz-f1 activates the expression of the steroid hormone receptor EcR-B1, a key component for γ remodelling, and represses expression of Hr39, an ftz-f1 homologous gene. If ectopically expressed in the γ neurons, HR39 inhibits normal pruning, probably by competing with endogenous FTZ-F1, which results in decreased EcR-B1 expression. The mushroom bodies are a bilaterally symmetric structure in the larval and adult brain and are involved in the processing of different types of olfactory memory. How memory is affected in pruning-deficient adult flies that possess larval-stage neuronal circuitry will help to explain the functional role of neuron remodelling. Flies overexpressing Hr39 are viable as adults and make it possible to assess the requirement for wild-type mushroom body pruning in memory. While blocking mushroom body neuron remodelling impaired memory after short-term courtship conditioning, long-term memory was normal. These results show that larval pruning is necessary for adult memory and that expression of courtship short-term memory and long-term memory may be parallel and independent.

Mushroom body neuronal remodelling is necessary for short-term but not for long-term courtship memory in Drosophila.

One of the challenges facing memory research is to combine network- and cellular-level descriptions of memory encoding. In this context, Drosophila offers the opportunity to decipher, down to single-cell resolution, memory-relevant circuits in connection with the mushroom bodies (MBs), prominent structures for olfactory learning and memory. Although the MB-afferent circuits involved in appetitive learning were recently described, the circuits underlying appetitive memory retrieval remain unknown. We identified two pairs of cholinergic neurons efferent from the MB α vertical lobes, named MB-V3, that are necessary for the retrieval of appetitive long-term memory (LTM). Furthermore, LTM retrieval was correlated to an enhanced response to the rewarded odor in these neurons. Strikingly, though, silencing the MB-V3 neurons did not affect short-term memory (STM) retrieval. This finding supports a scheme of parallel appetitive STM and LTM processing.

Two pairs of mushroom body efferent neurons are required for appetitive long-term memory retrieval in Drosophila.

In Drosophila, prior fighting experience influences the outcome of later contests: losing a fight increases the probability of losing second contests, thereby revealing "loser" effects that involve learning and memory. In these experiments, to generate and quantify the behavioral changes observed as consequences of losing fights, we developed a new behavioral arena that eliminates handling. We compared two commonly used fly handling procedures with this new chamber and demonstrated that handling influences aggressive behavior and prevents "loser" effect formation. In addition, we induced and observed novel aspects of learning associated with aggression such as the formation of robust winner effects.

Handling alters aggression and "loser" effect formation in Drosophila melanogaster.

Aggressive behavior in Drosophila melanogaster serves to acquire or defense vital resources such as food, territory or access to mates. Flies learn from previous fighting experience and modify and adapt their behavior to new situations, suggesting that learning and memory play a major role in agonistic encounters. Prior fighting experience influences the outcome of later contests: losing a fight increases the probability of losing second contests, revealing the formation of a "loser" effect. In a recent publication, we developed a new behavioral arena that eliminates handling of flies prior to, during and after fights to study the learning and memory associated with aggression. We compared two handling procedures commonly used in laboratories to study aggression with the new chambers and demonstrated that handling negatively influences aggression and prevents "loser" effect formation. In addition, we observed new aspects of behavior such as the formation of robust winner effects.

Learning and memory during aggression in Drosophila: handling affects aggression and the formation of a "loser" effect.

Understanding how the various memory components are encoded and how they interact to guide behavior requires knowledge of the underlying neural circuits. Currently, aversive olfactory memory in Drosophila is behaviorally subdivided into four discrete phases. Among these, short- and long-term memories rely, respectively, on the γ and α/β Kenyon cells (KCs), two distinct subsets of the ∼2,000 neurons in the mushroom body (MB). Whereas V2 efferent neurons retrieve memory from α/β KCs, the neurons that retrieve short-term memory are unknown. We identified a specific pair of MB efferent neurons, named M6, that retrieve memory from γ KCs. Moreover, our network analysis revealed that six discrete memory phases actually exist, three of which have been conflated in the past. At each time point, two distinct memory components separately recruit either V2 or M6 output pathways. Memory retrieval thus features a dramatic convergence from KCs to MB efferent neurons.

Article	Year
Mutation in TET2 in myeloid cancers. The New England journal of medicine\| PubMed ID: 19474426	2009
The steroid hormone receptor EcR finely modulates Drosophila lifespan during adulthood in a sex-specific manner. Mechanisms of ageing and development\| PubMed ID: 19486910	2009
Mushroom body efferent neurons responsible for aversive olfactory memory retrieval in Drosophila. Nature neuroscience\| PubMed ID: 21685917	2011
Parallel processing of appetitive short- and long-term memories in Drosophila. Current biology : CB\| PubMed ID: 21962716	2011
Slow oscillations in two pairs of dopaminergic neurons gate long-term memory formation in Drosophila. Nature neuroscience\| PubMed ID: 22366756	2012
Mushroom body neuronal remodelling is necessary for short-term but not for long-term courtship memory in Drosophila. The European journal of neuroscience\| PubMed ID: 22571719	2012
Two pairs of mushroom body efferent neurons are required for appetitive long-term memory retrieval in Drosophila. Cell reports\| PubMed ID: 24209748	2013
Handling alters aggression and "loser" effect formation in Drosophila melanogaster. Learning & memory (Cold Spring Harbor, N.Y.)\| PubMed ID: 25593291	2015
Learning and memory during aggression in Drosophila: handling affects aggression and the formation of a "loser" effect. Journal of nature and science\| PubMed ID: 25789340	2015
Two independent mushroom body output circuits retrieve the six discrete components of Drosophila aversive memory. Cell reports\| PubMed ID: 25981036	2015

Severine Trannoy

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