A behavioral and genetic dissection of two forms of olfactory plasticity in Caenorhabditis elegans: adaptation and habituation

Continuous presentation of an olfactory stimulus causes a decrement of the chemotaxis response in the nematode Caenorhabditis elegans. However, the differences between the learning process of habituation (a readily reversible decrease in behavioral response) and other types of olfactory plasticity such as adaptation (a decrement in response due to sensory fatigue, which cannot be dishabituated) have not been addressed. The volatile odorant diacetyl (DA) was used within a single paradigm to assess the distinct processes of olfactory adaptation and habituation. Preexposing and testing worms to 100% DA vapors caused a chemotaxis decrement that was not reversible despite the presentation of potentially dishabituating stimuli. This DA adaptation was abolished in worms with an odr-10 mutation (encoding a high-affinity DA receptor on the AWA neuron), even though naive chemotaxis remained unaffected. Conversely, DA adaptation remained intact in odr-1 mutants (defective in AWC neuron-mediated olfactory behavior), even though naive chemotaxis to DA decreased. Surprisingly, exposure to vapors of intermediate concentrations of DA (0.01% and 25%) did not cause worms to exhibit any response decrement. In contrast to preexposure to high DA concentrations, preexposure to low DA concentrations (0.001%) produced habituation of the chemotaxis response (a dishabituating stimulus could reverse the response decrement back to baseline levels). The distinct behavioral effects produced by DA preexposure highlight a concentration-dependent dissociation between two decremental olfactory processes: adaptation at high DA concentrations versus habituation at low DA concentrations.     

Protection from premature habituation requires functional mushroom bodies in Drosophila

Diminished responses to stimuli defined as habituation can serve as a gating mechanism for repetitive environmental cues with little predictive value and importance. We demonstrate that wild-type animals diminish their responses to electric shock stimuli with properties characteristic of short- and long-term habituation. We used spatially restricted abrogation of neurotransmission to identify brain areas involved in this behavioral response. We find that the mushroom bodies and, in particular, the α/β lobes appear to guard against habituating prematurely to repetitive electric shock stimuli. In addition to protection from premature habituation, the mushroom bodies are essential for spontaneous recovery and dishabituation. These results reveal a novel modulatory role of the mushroom bodies on responses to repetitive stimuli in agreement with and complementary to their established roles in olfactory learning and memory.     

Distinct neural mechanisms mediate olfactory memory formation at different timescales

Habituation is one of the oldest forms of learning, broadly expressed across sensory systems and taxa. Here, we demonstrate that olfactory habituation induced at different timescales (comprising different odor exposure and intertrial interval durations) is mediated by different neural mechanisms. First, the persistence of habituation memory is greater when mice are habituated on longer timescales. Second, the specificity of the memory (degree of cross-habituation to similar stimuli) also depends on induction timescale. Third, we demonstrate a pharmacological double dissociation between the glutamatergic mechanisms underlying short- and long-timescale odor habituation. LY341495, a class II/III metabotropic glutamate receptor antagonist, blocked habituation only when the induction timescale was short. Conversely, MK-801, an N-methyl-D-aspartate (NMDA) receptor antagonist, prevented habituation only when the timescale was long. Finally, whereas short-timescale odor habituation is mediated within the anterior piriform cortex, infusion of MK-801 into the olfactory bulbs prevented odor habituation only at longer timescales. Thus, we demonstrate two neural mechanisms underlying simple olfactory learning, distinguished by their persistence and specificity, mediated by different olfactory structures and pharmacological effectors, and differentially utilized based solely on the timescale of odor presentation.     

Regulation of distinct attractive and aversive mechanisms mediating benzaldehyde chemotaxis in Caenorhabditis elegans

Olfactory-mediated chemotaxis in nematodes provides a relatively simple system to study biological mechanisms of information processing. Analysis of the kinetics of chemotaxis in response to 100% benzaldehyde revealed an initial attractive response that is followed by a strong aversion to the odorant. We show that this behavior is mediated by two genetically separable attraction- and aversion-mediating response pathways. The attraction initially dominates behavior but with prolonged exposure habituation leads to a behavioral change, such that the odorant becomes repulsive. This olfactory habituation is susceptible to dishabituation, thereby re-establishing the attractive response to the odorant. Re-examination of the putative olfactory adaptation mutant adp-1(ky20) revealed that the phenotype observed in this line is due to a supersensitivity to a dishabituating stimulus, rather than a defect in the adaptation to odorants per se. A modified benzaldehyde chemotaxis assay was developed and used for the isolation of a mutant with a specific defect in habituation kinetics, expressed as a persistence of the attractive response.     

Contextual control of long-term habituation in rats

This study examined contextual control of long-term habituation and whether such effects are dependent on the habituating response system. Habituation of the acoustic startle response transferred from the home cage to the testing context, whereas habituation of lick suppression was context specific (Experiments 1 and 2). Contextual control of habituation was demonstrated between 2 experimental contexts for lick suppression to a tone (Experiment 3) and bar-press suppression to a light (Experiment 4). Experiment 5 extinguished habituation of lick suppression and the orienting response to a tone with 27 exposures to the habituation context. Context specificity of both responses also was found. Previous failures to demonstrate contextual control of habituation may be due to the choice of response system and to less sensitive procedures to detect response recovery. The habituation mechanism for startle is independent from the process or processes that underlie habituation in other response systems, but the nature of these mechanisms is not yet known.     

Long-term habituation to spatial novelty in blind cave fish (Astyanax hubbsi): role of the telencephalon and its subregions

Blind cave fish, when released into a novel environment, show a typical exploratory behavior characterized by high swim speed along walls shortly after release. This behavior wanes during prolonged exposure and thus may reflect habituation to novelty. As the hippocampus of mammals, which plays a crucial role in spatial learning, is part of the telencephalon, the possible involvement of this brain structure of fish was investigated in exploratory behavior. Ablation of the whole telencephalon or bilateral removal of dorsal parts of the hemispheres reduced activity; in contrast, unilateral lesions of one hemisphere, bilateral lesions of dorsal and dorsoventral parts, and removal of olfactory bulbs increased activity. However, the time course of habituation in a novel environment remained unchanged, except for ablated animals in which there was virtually no habituation in swim speed. These data suggest that the telencephalon of fish may not participate in long-term habituation to spatial novelty but, rather, support the notion of the telencephalon being involved in generation of arousal.     

An exploratory investigation of coffee and lemon scents and odor identification

Fragrance sellers often provide coffee beans to their customers as a "nasal palate cleanser," to reduce the effects of olfactory adaptation and habituation. To test this idea, college students smelled three fragrances multiple times, rating odors each time. After completing nine trials, participants sniffed coffee beans, lemon slices, or plain air. Participants then indicated which of four presented fragrances had not been previously smelled; Coffee beans did not yield better performance than lemon slices or air.     

Heart-rate and electrodermal orienting responses to visual stimuli differing in complexity

Orienting response (OR) theory predicts that amount of information in the stimulus, or stimulus complexity, should be an important determinant of OR elicitation and habituation, more intense and more slowly habituating ORs being expected to complex than to simple stimuli. This prediction was tested in an experiment where subjects were exposed to simple and complex visual stimuli in randomized order, while heart-rate and skin conductance were measured. Complex stimuli evoked a more pronounced deceleratory heartrate response than did simple stimuli. However, the two conditions did not differ in rate of habituation of this response. For skin conductance responses, on the other hand, the complex stimulus took more trials than the simple stimulus to reach habituation, whereas the two conditions did not differ in response magnitude. Thus, the hypothesis of more intense orienting to complex stimuli was supported by the heart-rate data, and that of slower habituation to complex stimuli, of the skin conductance data.     

Extraversion and neuroticism in orienting reflex dishabituation

The present study examined the effects of extraversion (E) and neuroticism (N) as individual difference dimensions and of stimulus intensity on electrodermal orienting response habituation, test response, and dishabituation. Subjects were 162 college students selected from a screening sample of 860 as representing extreme groups of high, medium, and low E crossing with high, medium, and low N. Subjects in all nine resulting groups were habituated to criterion on auditory stimuli of 60, 80, and 100 db. A novel (test) stimulus and a dishabituating standard stimulus followed criteron habituation at each intensity. Results indicated that response amplitude for initial, test, and dishabituation responses was a direct function and that habituation rate (trials-to-criterion) was an inverse function of stimulus intensity. Extraverts habituated more rapidly than introverts at high intensity and gave smaller test and dishabituation responses. Some results differed when regression coefficients defined habituation rate and when first stimulus intensities alone were examined. Correlations among initial, test, and dishabituation responses and between each of these and the two measures of habituation rate are also reported. Results were interpreted as partially supportive of Eysenck's theory.     

Effects of stimulus duration and intensity on the rate and specificity of behavioural habituation

Tone stimuli were presented to rats once per day until habituation was reached. Neither the duration nor the intensity of the stimulus affected the rate of habituation. Habituation was specific to both these stimulus parameters as shown by the distraction produced by either an increase or a decrease in the original stimulus value. It is suggested that some of the discrepancies in the literature might be resolved if a distinction is drawn between habituation of orienting responses and the decrement of more specific responses.     

Synaptic adaptation and odor-background segmentation

Habituation is a form of non-associative memory that plays an important role in filtering stable or redundant inputs. The present study examines the contribution of habituation and cortical adaptation to odor-background segmentation. Segmentation of target odorants from background odorants is a fundamental computational requirement for the olfactory system. Recent electrophysiological data have shown that odor specific adaptation in piriform cortex neurons, mediated at least partially by synaptic adaptation between the olfactory bulb outputs and piriform cortex pyramidal cells, may provide an ideal mechanism for odor-background segmentation. This rapid synaptic adaptation acts as a filter to enhance cortical responsiveness to changing stimuli, while reducing responsiveness to static, potentially background stimuli. Using previously developed computational models of the olfactory system, we here show how synaptic adaptation at the olfactory bulb input to the piriform cortex, as demonstrated electrophysiologically, creates odor specific adaptation. We show how this known feature of olfactory cortical processing can contribute to adaptation to a background odor and to odor-background segmentation. We then show in a behavioral experiment that the odor-background segmentation is perceptually important and functions at the same time-scale as the synaptic adaptation observed between the olfactory bulb and cortex.     

Auditory habituation in the fetus and neonate: an fMEG study

Habituation – the most basic form of learning – is used to evaluate central nervous system (CNS) maturation and to detect abnormalities in fetal brain development. In the current study, habituation, stimulus specificity and dishabituation of auditory evoked responses were measured in fetuses and newborns using fetal magnetoencephalography (fMEG). An auditory habituation paradigm consisting of 100 trains of five 500 Hz tones, one 750 Hz tone (dishabituator) and two more 500 Hz tones, respectively, were presented to 41 fetuses (gestational age 30–39 weeks) and 22 newborns or babies (age 6–89 days). A response decrement between the first and fifth tones (habituation), an increment between the fifth tone and the dishabituator (stimulus specificity) and an increment between the fifth (last tone before the dishabituator) and seventh tones (first tone after the dishabituator) (dishabituation) were expected. Fetuses showed weak responses to the first tone. However, a significant response decrement between the second and fifth tones (habituation) and a significant increment between the fifth tone and the dishabituator (stimulus specificity) were found. No significant difference was found for dishabituation nor was a developmental trend found at the group level. From the neonatal data, significant values for stimulus specificity were found. Sensory fatigue or adaptation was ruled out as a reason for the response decrement due to the strong reactions to the dishabituator. Taken together, the current study used fMEG to directly show fetal habituation and provides evidence of fetal learning in the last trimester of pregnancy.     

Napping promotes inter-session habituation to emotional stimuli

The effects of a daytime nap on inter-session habituation to aversive visual stimuli were investigated. Healthy young adult volunteers viewed repeated presentations of highly negative and emotionally neutral (but equally arousing) International Affective Picture System (IAPS) photographs during two afternoon sessions separated by 2.5h. Half of the photographs were shown at both sessions (Repeated Sets) and half differed between sessions (Novel Sets). For each stimulus presentation, evoked skin conductance response (SCR), heart-rate deceleration (HRD) and corrugator supercilii EMG response (EMG), were computed and range corrected using respective maximum session-1 responses. Following each presentation, subjects rated each photograph on dimensions of pleasantness and arousability. During the inter-session interval, Nap subjects had a 120-min polysomnographically monitored sleep opportunity, whereas Wake subjects watched a non-stimulating video. Nap and Wake subjects did not differ in their subjective ratings of photographs. However, for Repeated-Set photographs, Nap subjects demonstrated greater inter-session habituation in SCR and EMG but a trend toward lesser inter-session habituation in HRD. These group differences were absent for Novel-Set photographs. Group differences across all measures were greater for negative stimuli. Occurrence of SWS during the nap was associated with greater inter-session habituation of EMG whereas occurrence of REM was associated with lesser inter-session habituation of SCR to negative stimuli. Sleep may therefore promote emotional adjustment at the level of somatic responses. Physiological but not subjective inter-session habituation to aversive images was enhanced by a daytime nap.     

Skin conductance,reaction time,and observational ratings in learning-disabled children

Rates of GSR habituation, reactiontime responses, and observational ratings in normal and learningdisabled children were evaluated. Compared to normals, children with learning disabilities demonstrated faster rates of GSR habituation in a task involving active attention, slower reactiontime responses, and more characteristics of “minimal brain dysfunction” on a rating scale. Within the learningdisability group those subjects with a family history of reading problems showed fewer GSR, reactiontime, and rating responses characteristic of “minimal brain dysfunction.”     

The sensitization of humor responses to cartoons

It was hypothesized that a humor response to a cartoon in a series depends upon arousal or increased salience, both of which may be produced by preceding cartoons. A humor response, however, should eventually decline owing to habituation. Three experiments were conducted in order to determine if responding to cartoons with the same theme depends upon the cartoon's serial position. Facial responses and ratings of funniness increased over the first few cartoons, reflecting the effect of arousal or salience. It appears that the initial cartoons make the person more sensitive to later cartoons. A downward trend in responding was more apparent for facial responses than for funniness ratings. The evidence for habituation was weak, however, and did not seem occur to the experimenter-defined cartoon theme. Cartoons even representing the same theme may be too variable to permit habituation. It was suggested that cartoon researchers be aware of the effect of a cartoon's serial position on humor responses.The investigators would like to thank Michael Davis, Michael Mullins, and Elizabeth Weaver for their assistance in data collection and analyses.     

Behavioural arousal and its habituation in the squirrel fish, Holocentrus rufus: the role of the telecephalon

Squirrel fish were captured, maintained for 2 days and then tested for their responses to stimuli in laboratory aquaria. A tap on the side of the aquarium or a shadow moving overhead elicited an arousal response involving a reduction in opercular movements, movement of the pectoral fins, and erection of the spiny dorsal fin. This latter response, when regularly provoked by a moving shadow at 15-s intervals, decreased in duration and habituated. When fish were restricted in a wire mesh trough and the stimulus was the onset of illumination to one eye at 15-s intervals, responses were quantitatively similar although shorter in duration. A series of stimuli delivered to one eye until habituation followed by a series to the other eye reduced the duration of initial responses and the number of trials for habituation of responses to stimuli delivered to the second eye. Responses were equivalent regardless of which eye (left or right) was first presented with stimuli. Section of the telencephalic anterior commissure considerably restricted the transfer of information about experience of stimuli presented to the first eye. Commissural section, combined with unilateral lesions in the posterior-medial telencephalon, severely impaired habituation of responses to the contralateral eye. Lateral lesions had no effect. The results are discussed in relation to the visual projection in Holocentrus and telecephalic involvement in arousal and its habituation in teleosts as a whole.     

Olfactory aversion conditioning and overeating: a review and some data

The present study evaluated the efficacy of olfactory aversion conditioning in the management of overeating problems. 42 overweight female subjects were assigned to one of three treatment conditions: olfactory aversion therapy, attention placebo control, and no-contact control. One experimenter administered the 8-wk. treatment phase. The aversion therapy procedure entailed the pairing of selected target foods (CSs) with noxious odors (UCSs). There were 25 pairings of the CS and UCSs during each weekly session. Four noxious odors were employed, one each week, to prevent habituation to the UCS. The attention-placebo control procedure was identical except that "air" was substituted for the putative UCS of the aversion therapy condition. At the end of the treatment period the aversion therapy group had lost 4.7 lb.; the attention placebo controls had lost 3.6 lb. and the no-contact controls 0.5 lb. The difference between the aversion therapy group and the no-contact controls was significant and that between the attention placebo group and the no-contact controls approached significance. At a follow-up 8 wk. after the end of the treatment period the weights of all groups had risen to pretreatment levels and there were no differences between them. These results indicate that olfactory aversion therapy is not an efficient technique in promoting weight-loss.     

Habituation in songbirds

Songbirds respond to initial playback of a recorded conspecific song in numerous ways, from changes in gene expression in the brain to changes in overt physical activity. When the same song is presented repeatedly, responses have been observed to habituate at multiple levels: molecular, cellular and organismal. Core criteria of habituation have been established at each level, although in no case have all the formal parameters been rigorously measured. At the level of overt behavior, classical field studies showed that territorial birds respond to the song of a potential challenger with a variety of behaviors, and many (but not all) of these behaviors decline with repeated stimulus presentation. More recent laboratory studies have defined analogous responses to song presentation in the zebra finch (Taeniopygia guttata), the dominant species in current molecular and neurobiological research and one that does not use song for territorial defense. Studies in the zebra finch have also demonstrated activation followed by habituation of responses measured at both electrophysiological and molecular (gene expression and signal transduction) levels. In all cases, habituation is specific for a very particular stimulus – an individual song presented in a particular context. There are strong correlations between habituation measurements made at these different levels, but some dissociations have also been observed, implying that molecular, electrophysiological and behavioral habituations are not equivalent manifestations of a single core process.     

Odor-specific habituation arises from interaction of afferent synaptic adaptation and intrinsic synaptic potentiation in olfactory cortex

Segmentation of target odorants from background odorants is a fundamental computational requirement for the olfactory system and is thought to be behaviorally mediated by olfactory habituation memory. Data from our laboratory have shown that odor-specific adaptation in piriform neurons, mediated at least partially by synaptic adaptation between the olfactory bulb outputs and piriform cortex pyramidal cells, is highly odor specific, while that observed at the synaptic level is specific only to certain odor features. Behavioral data show that odor habituation memory at short time constants corresponding to synaptic adaptation is also highly odor specific and is blocked by the same pharmacological agents as synaptic adaptation. Using previously developed computational models of the olfactory system we show here how synaptic adaptation and potentiation interact to create the observed specificity of response adaptation. The model analyzes the mechanisms underlying the odor specificity of habituation, the dependence on functioning cholinergic modulation, and makes predictions about connectivity to and within the piriform neural network. Predictions made by the model for the role of cholinergic modulation are supported by behavioral results.Filtering sensory input is critical for information processing tasks such as background segmentation, and shifting processing power away from redundant, stable, or repetitive stimuli toward dynamic, novel stimuli. A critical aspect of this filtering however, is stimulus specificity. Under most circumstances it may be most beneficial to selectively filter the redundant stimulus, while maintaining responsiveness to different, though perhaps highly similar stimuli.In the olfactory system, short-term habituation to stable or repeated odorants involves a metabotropic glutamate receptor (mGluR)-dependent depression of afferent synapses to the piriform cortex (Best and Wilson 2004). Blockade of group III mGluR receptors prevents cortical adaptation odors (Best and Wilson 2004), and reduces short-term habituation of odor-evoked reflexes (Best et al. 2005) and odor investigation (Yadon and Wilson 2005; Bell et al. 2008; McNamara et al. 2008). This short-term habituation is highly odor specific, with minimal cross-adaptation of piriform cortical single-unit responses or cross-habituation of behavioral responses to similar odors, including between mixtures and their components (Wilson 2000; Cleland et al. 2002). Interestingly, there is an experience-dependent component to short-term habituation odor specificity. The odor specificity is most pronounced for familiar odors, with very brief (<20 sec) exposure to odors producing more generalization, and longer exposures (>50 sec) sufficient to permit strong odor specificity in cortex adaptation (Wilson 2003).The homosynaptic nature of afferent synaptic depression underlying cortical adaptation (Wilson 1998; Best and Wilson 2004) may contribute to this odor specificity. However, the experience dependence suggests that there may be an additional process involved. In fact, theoretical views of piriform cortical function suggest that the cortex learns previous patterns of input via potentiation of intracortical association fiber synapses (Hasselmo et al. 1990; Barkai et al. 1994; Haberly 2001; Linster et al. 2003). This autoassociative process essentially creates a template of previous network activity, against which new input patterns can be compared, allowing enhanced discrimination between similar patterns, as well as completion of degraded patterns (Barkai et al. 1994; Barnes et al. 2008). In support of this hypothesis, previous work has demonstrated that disruption of normal synaptic potentiation in association fiber synapses through blockade of cholinergic muscarinic receptors (Patil et al. 1998; Linster et al. 2003), reduces odor specificity of cortical adaptation (Wilson 2001b), prevents the effects of odor experience on subsequent behavioral cross-habituation (Fletcher and Wilson 2002), and disrupts odor discrimination (Linster et al. 2001).The present series of studies further explored the role of combined afferent synaptic depression and intracortical association fiber synaptic potentiation on the specificity of cortical adaptation and odor habituation. Using a computational model of the olfactory system (Linster et al. 2007), the results suggest that activity-dependent association fiber plasticity is necessary to account for the specificity of odor habituation. Furthermore, in behavioral experiments blockade of cholinergic muscarinic receptors during habituation enhances generalization of odor habituation, consistent with the modeling and with previous electrophysiological results.     

Fetal habituation performance: Gestational age and sex effects

Habituation is the decrement in response to repeated stimulation. Fetal habituation performance may reflect the functioning of the central nervous system (CNS) prenatally. However, basic characteristics of the prenatal habituation phenomena remain unclear, such as the relationship with gestational age (GA) and fetal sex. The current study investigated the relationship between habituation performance and both fetal sex and advancing gestational age, by comparing habituation performance at 31 and 35 weeks GA within a sample of 74 singleton pregnancies, all with normal neonatal outcomes. A vibroacoustic stimulus was used to habituate the fetus and motor responses were monitored via ultrasound. The criterion for habituation was defined as no response to five consecutive stimulus presentations. Prenatal habituation performance was found to demonstrate a developmental trend, with an increased proportion of fetuses habituating with advancing gestational age and requiring fewer stimulus presentations to satisfy the habituation criterion. Fetal sex also appears to have a moderating influence on gestational changes in habituation performance. Hence, the ongoing maturation of the fetal CNS does appear to be reflected in fetal habituation performance, indicating the need to consider the composition of fetal samples in terms of both gestational age and sex. Variability in interfetal performance was found to be high and clinical applications remain speculative.