Use of an Immediate Swallow Protocol to Assess Taste and Aroma Integration in fMRI Studies

Perception of flavor is a complex process involving the integration of taste and aroma. Few functional magnetic resonance imaging (fMRI) studies have assessed the crossmodal interactions which result in flavor perception, and all previous studies have used a retro-nasal aroma delivery with a delayed swallow, which delays retro-nasal aroma release, and thus, alters taste and aroma integration. In this paper, we assess crossmodal interactions in flavor processing using an immediate swallow fMRI paradigm in 13 healthy volunteers. We compare unimodal taste (sucrose) and unimodal retro-nasal aroma stimuli, with a congruent taste and aroma combination (flavor), to assess crossmodal flavor interactions using an immediate swallow paradigm. Subtraction and conjunction analysis methods are described, and the use of a control stimulus is addressed. Subtraction analysis was found to reveal areas of anterior insula, frontal operculum, anterior cingulate, and orbitofrontal cortex, whilst the conjunction analysis revealed additional active areas in oral somatosensory areas (SI), rolandic operculum and posterior cingulate, supporting the hypothesis that taste, olfactory, and tactile sensations are integrated to produce a flavor percept.     

Taste-olfactory convergence, and the representation of the pleasantness of flavour, in the human brain

The functional architecture of the central taste and olfactory systems in primates provides evidence that the convergence of taste and smell information onto single neurons is realized in the caudal orbitofrontal cortex (and immediately adjacent agranular insula). These higher-order association cortical areas thus support flavour processing. Much less is known, however, about homologous regions in the human cortex, or how taste-odour interactions, and thus flavour perception, are implemented in the human brain. We performed an event-related fMRI study to investigate where in the human brain these interactions between taste and odour stimuli (administered retronasally) may be realized. The brain regions that were activated by both taste and smell included parts of the caudal orbitofrontal cortex, amygdala, insular cortex and adjoining areas, and anterior cingulate cortex. It was shown that a small part of the anterior (putatively agranular) insula responds to unimodal taste and to unimodal olfactory stimuli, and that a part of the anterior frontal operculum is a unimodal taste area (putatively primary taste cortex) not activated by olfactory stimuli. Activations to combined olfactory and taste stimuli where there was little or no activation to either alone (providing positive evidence for interactions between the olfactory and taste inputs) were found in a lateral anterior part of the orbitofrontal cortex. Correlations with consonance ratings for the smell and taste combinations, and for their pleasantness, were found in a medial anterior part of the orbitofrontal cortex. These results provide evidence on the neural substrate for the convergence of taste and olfactory stimuli to produce flavour in humans, and where the pleasantness of flavour is represented in the human brain.     

Superadditive opercular activation to food flavor is mediated by enhanced temporal and limbic coupling

Food perception is characterized by a transition from initially separate sensations of the olfactory and gustatory properties of the object toward their combined sensory experience during consumption. The holistic flavor experience, which occurs as the smell and taste merge, extends beyond the mere addition of the two chemosensory modalities, being usually perceived as more object‐like, intense and rewarding. To explore the cortical mechanisms which give rise to olfactory–gustatory binding during natural food consumption, brain activation during consumption of a pleasant familiar beverage was contrasted with presentation of its taste and orthonasal smell alone. Convergent activation to all presentation modes was observed in executive and chemosensory association areas. Flavor, but not orthonasal smell or taste alone, stimulated the frontal operculum, supporting previous accounts of its central role in the formation of the flavor percept. A functional dissociation was observed in the insula: the anterior portion was characterized by sensory convergence, while mid‐dorsal sections activated exclusively to the combined flavor stimulus. psycho‐physiological interaction analyses demonstrated increased neural coupling between the frontal operculum and the anterior insula during flavor presentation. Connectivity was also increased with the lateral entorhinal cortex, a relay to memory networks and central node for contextual modulation of olfactory processing. These findings suggest a central role of the insular cortex in the transition from mere detection of chemosensory convergence to a superadditive flavor representation. The increased connections between the frontal operculum and medial temporal memory structures during combined olfactory–gustatory stimulation point to a potential mechanism underlying the acquisition and modification of flavor preferences. Hum Brain Mapp 36:1662–1676, 2015. © 2014 Wiley Periodicals, Inc.     

The Representation of Information About Taste and Odor in the Orbitofrontal Cortex

Complementary neurophysiological recordings in macaques and functional neuroimaging in humans show that the primary taste cortex in the rostral insula and adjoining frontal operculum provides separate and combined representations of the taste, temperature, and texture (including viscosity and fat texture) of food in the mouth independently of hunger and thus of reward value and pleasantness. One synapse on, in the orbitofrontal cortex, these sensory inputs are for some neurons combined by learning with olfactory and visual inputs, and these neurons encode food reward in that they only respond to food when hungry and in that activations here correlate with subjective pleasantness and with individual differences in and cognitive modulation of the hedonic value of food. Information theory analysis shows a robust representation of taste in the orbitofrontal cortex, with an average mutual information of 0.45 bits for each neuron about which of six tastants (glucose, NaCl, HCl, quinine-HCl, monosodium glutamate, and water) was present, averaged across 135 gustatory neurons. The information increased with the number of neurons in the ensemble, but less than linearly, reflecting some redundancy. There was less information per neuron about which of six odors was present from orbitofrontal olfactory neurons, but the code was robust in that the information increased linearly with the number of neurons, reflecting independent information encoded by different neurons. Although some neurons were sharply tuned to individual tastants, the average encoding was quite distributed.     

Swallowing intentional off-state in aging and Alzheimer's disease: preliminary study

Frontal cortical activation is elicited when subjects have been instructed not to initiate a sensorimotor task. The goal of this preliminary fMRI study was to examine BOLD response to a "Do Not Swallow" instruction (an intentional "off-state") in the context of other swallowing tasks in 3 groups of participants (healthy young, healthy old, and early Alzheimer's disease (AD)). Overall, the older group had larger, bilaterally active clusters in the cortex, including the dorsomedial prefrontal cortex during the intentional swallowing off-state; this region is commonly active in response inhibition studies. Disease-related differences were evident where the AD group had significantly greater BOLD response in the insula/operculum than the old. These findings have significant clinical implications for control of swallowing across the age span and in neurodegenerative disease. Greater activation in the insula/operculum for the AD group supports previous studies where this region is associated with initiating swallowing. The AD group may have required more effort to "turn off" swallowing centers to reach the intentional swallowing off-state.     

A bold view of the lactating brain: functional magnetic resonance imaging studies of suckling in awake dams

Functional magnetic resonance imaging (fMRI) has been used to investigate the responsiveness of the maternal rat brain to pup-suckling under various experimental paradigms. Our research employing the lactating rat model has explored the cortical sensory processing of pup stimuli and the effect of suckling on the brain's reward system. Suckling was observed to increase blood-oxygen-level-dependent (BOLD) signal intensity in the midbrain, striatum and prefrontal cortex, which are areas that receive prominent dopaminergic inputs. The BOLD activation of the reward system occurs in parallel with the activation of extensive cortical sensory areas. The observed regions include the olfactory cortex, auditory cortex and gustatory cortex, and could correspond to cortical representations of pup odours, vocalisations and taste that are active during lactation. Activation patterns within reward regions are consistent with past research on maternal motivation and we explore the possibility that exposure to drugs of abuse might be disruptive of maternal neural responses to pups, particularly in the prefrontal cortex. Our ongoing fMRI studies support and extend past research on the maternal rat brain and its functional neurocircuitry.     

Impact of Olfaction on Taste, Trigeminal, and Texture Perceptions

Research on perceptual interactions has mainly been conducted on olfaction and taste. Few studies have extended the investigation to trigeminal and texture perceptions. The objective of the study was to explore systematical interactions between olfaction, taste, texture, and trigeminal perceptions with a specific focus on the role of olfaction. Ingredients inducing olfactory (odorant), taste (citric acid), and trigeminal (cooling agent) perceptions were systematically varied in a viscous model system. A panel assessed sensory properties of the products within the same odorant type under two different conditions: with and without noseclip. Olfaction strongly influenced taste and trigeminal perceptions but also modulated perceptual taste/taste and taste/trigeminal interactions. But results involving texture perception were inconclusive since the evaluation condition (with and without wearing a noseclip) might have impacted mastication and swallowing behaviors. These findings highlight the multiplicity and overlapping of olfactory/trigeminal/taste perceptual interactions in complex food systems.     

Chemosensory processing in children with attention-deficit/hyperactivity disorder

BackgroundIn attention-deficit/hyperactivity disorder (ADHD) not only deficits in dopamine-related cognitive functioning have been found but also a lower dopamine-sensitive olfactory threshold. The aim of the present study was to proof that only olfactory but not trigeminal sensitivity is increased in ADHD. Structural magnetic resonance imaging (MRI) was used to show increased olfactory bulb (OB) volume- a structure which is strongly shaped by olfactory performance through the mechanism of neuroplasticity (e.g. synaptogenesis). To elucidate whether cortical mechanisms are involved in altered olfaction in ADHD, functional MRI (fMRI) was introduced.MethodsA total of 18 boys with ADHD and 17 healthy controls (aged 7–12) were included in the study. Olfactory as well as trigeminal detection thresholds were examined. OB sizes were measured by means of structural MRI and an analysis of effective functional (fMRI) coupling of primary olfactory cortex was conducted. The frontal piriform cortex (fPIR) was chosen as seed region because of its importance in processing both trigeminal and olfactory stimuli as well as having profound influence on inner OB-signaling.ResultsIncreased olfactory sensitivity as well as an increase in OB volume was found in ADHD. There were no group differences in sensitivity towards a trigeminal stimulus. Compared to healthy controls, the fPIR in ADHD was more positively coupled with structures belonging to the salience network during olfactory and, to a lesser extent, during trigeminal stimulation.ConclusionsOlfactory functioning is superior in subjects with ADHD. The observed increase in OB volume may relate to higher olfactory sensitivity in terms of neuroplasticity. During the processing of chemosensory stimuli, the primary olfactory cortex in ADHD is differently coupled to higher cortical structures which might indicate an altered top-down influence on OB structure and function.     

Networks involved in olfaction and their dynamics using independent component analysis and unified structural equation modeling

The study of human olfaction is complicated by the myriad of processing demands in conscious perceptual and emotional experiences of odors. Combining functional magnetic resonance imaging with convergent multivariate network analyses, we examined the spatiotemporal behavior of olfactory‐generated blood‐oxygenated‐level‐dependent signal in healthy adults. The experimental functional magnetic resonance imaging (fMRI) paradigm was found to offset the limitations of olfactory habituation effects and permitted the identification of five functional networks. Analysis delineated separable neuronal circuits that were spatially centered in the primary olfactory cortex, striatum, dorsolateral prefrontal cortex, rostral prefrontal cortex/anterior cingulate, and parietal‐occipital junction. We hypothesize that these functional networks subserve primary perceptual, affective/motivational, and higher order olfactory‐related cognitive processes. Results provided direct evidence for the existence of parallel networks with top‐down modulation for olfactory processing and clearly distinguished brain activations that were sniffing‐related versus odor‐related. A comprehensive neurocognitive model for olfaction is presented that may be applied to broader translational studies of olfactory function, aging, and neurological disease. Hum Brain Mapp 35:2055–2072, 2014. © 2013 Wiley Periodicals, Inc .     

Activation of multiple cortical areas in response to somatosensory stimulation: combined magnetoencephalographic and functional magnetic resonance imaging.

We combined information from functional magnetic resonance imaging (fMRI) and magnetoencephalography (MEG) to assess which cortical areas and in which temporal order show macroscopic activation after right median nerve stimulation. Five healthy subjects were studied with the two imaging modalities, which both revealed significant activation in the contra- and ipsilateral primary somatosensory cortex (SI), the contra- and ipsilateral opercular areas, the walls of the contralateral postcentral sulcus (PoCS), and the contralateral supplementary motor area (SMA). In fMRI, two separate foci of activation in the opercular cortex were discerned, one posteriorly in the parietal operculum (PO), and one anteriorly near the insula or frontal operculum (anterior operculum, AO). The activation sites from fMRI were used to constrain the solution of the inverse problem of MEG, which allowed us to construct a model of the temporal sequence of activation of the different sites. According to this model, the mean onset latency for significant activation at the contralateral SI was 20 msec (range, 17-22 msec), followed by activation of PoCS at 23 msec (range, 21-25 msec). The contralateral PO was activated at 26 msec (range, 19-32 msec) and AO at 33 msec (range, 22-51 msec). The contralateral SMA became active at 36 msec (range, 24-48 msec). The ipsilateral SI, PO, and AO became activated at 54-67 msec. We conclude that fMRI provides a useful means to constrain the inverse problem of MEG, allowing the construction of spatiotemporal models of cortical activation, which may have significant implications for the understanding of cortical network functioning.     

Identifying human parieto-insular vestibular cortex using fMRI and cytoarchitectonic mapping

The parieto-insular vestibular cortex (PIVC) plays a central role in the cortical vestibular network. Although this region was first defined and subsequently extensively studied in nonhuman primates, there is also ample evidence for a human analogue in the posterior parietal operculum. In this study, we functionally and anatomically characterize the putative human equivalent to macaque area PIVC by combining functional magnetic resonance imaging (fMRI) of the cortical response to galvanic vestibular stimulation (GVS) with probabilistic cytoarchitectonic maps of the human parietal operculum. Our fMRI data revealed a bilateral cortical response to GVS in posterior parieto-insular cortex. Based on the topographic similarity of these activations to primate area PIVC, we suggest that they constitute the functionally defined human equivalent to macaque area PIVC. The locations of these activations were then compared to the probabilistic cytoarchitectonic maps of the parietal operculum (Eickhoff et al. [2005a]: Cereb Cortex, in press; Eickhoff et al. [2005c]: Cereb Cortex, in press), whereby the functionally defined PIVC matched most closely the cytoarchitectonically defined area OP 2. This activation of OP 2 by vestibular stimulation and its cytoarchitectonic features, which are similar to other primary sensory areas, suggest that area OP 2 constitutes the human equivalent of macaque area PIVC.     

Identification of human gustatory cortex by activation likelihood estimation

Over the last two decades, neuroimaging methods have identified a variety of taste-responsive brain regions. Their precise location, however, remains in dispute. For example, taste stimulation activates areas throughout the insula and overlying operculum, but identification of subregions has been inconsistent. Furthermore, literature reviews and summaries of gustatory brain activations tend to reiterate rather than resolve this ambiguity. Here, we used a new meta-analytic method [activation likelihood estimation (ALE)] to obtain a probability map of the location of gustatory brain activation across 15 studies. The map of activation likelihood values can also serve as a source of independent coordinates for future region-of-interest analyses. We observed significant cortical activation probabilities in: bilateral anterior insula and overlying frontal operculum, bilateral mid dorsal insula and overlying Rolandic operculum, and bilateral posterior insula/parietal operculum/postcentral gyrus, left lateral orbitofrontal cortex (OFC), right medial OFC, pregenual anterior cingulate cortex (prACC) and right mediodorsal thalamus. This analysis confirms the involvement of multiple cortical areas within insula and overlying operculum in gustatory processing and provides a functional "taste map" which can be used as an inclusive mask in the data analyses of future studies. In light of this new analysis, we discuss human central processing of gustatory stimuli and identify topics where increased research effort is warranted.     

An fMRI Study of the Interactions Between the Attention and the Gustatory Networks

In a prior study, we showed that trying to detect a taste in a tasteless solution results in enhanced activity in the gustatory and attention networks. The aim of the current study was to use connectivity analyses to test if and how these networks interact during directed attention to taste. We predicted that the attention network modulates taste cortex, reflecting top-down enhancement of incoming sensory signals that are relevant to goal-directed behavior. fMRI was used to measure brain responses in 14 subjects as they performed two different tasks: (1) trying to detect a taste in a solution or (2) passively perceiving the same solution. We used psychophysiological interaction analysis to identify regions demonstrating increased connectivity during a taste attention task compared to passive tasting. We observed greater connectivity between the anterior cingulate cortex and the frontal eye fields, posterior parietal cortex, and parietal operculum and between the anterior cingulate cortex and the right anterior insula and frontal operculum. These results suggested that selective attention to taste is mediated by a hierarchical circuit in which signals are first sent from the frontal eye fields, posterior parietal cortex, and parietal operculum to the anterior cingulate cortex, which in turn modulates responses in the anterior insula and frontal operculum. We then tested this prediction using dynamic causal modeling. This analysis confirmed a model of indirect modulation of the gustatory cortex, with the strongest influence coming from the frontal eye fields via the anterior cingulate cortex. In summary, the results indicate that the attention network modulates the gustatory cortex during attention to taste and that the anterior cingulate cortex acts as an intermediary processing hub between the attention network and the gustatory cortex.     

Cortical spatial aspects of optical intrinsic signals in response to sucrose and NaCl stimuli

To examine whether cortical taste neurons use spatial codes for discriminating taste information, we investigated the spatial aspects of optical intrinsic signal (OIS) responses in the gustatory insular cortex (GC) elicited by the administration of two essential tastants, sucrose and NaCl, on the tongue. OIS responses to sucrose appeared in the rostral part of the GC, whereas those to NaCl appeared in the central part of the GC. Local anesthetization of the tongue abolished OIS responses, and the administration of distilled water elicited no OIS response. Thus, taste information elicited by sucrose and NaCl from the peripheral sensory organs is segregated in the GC, suggesting that the information from two essential tastants is assembled as spatial codes in the primary cortical taste area through the process of taste quality perception.     

Methods for olfactory fMRI studies: Implication of respiration

Human olfactory system is under‐studied using fMRI compared with other sensory systems. Because the perception (intensity, threshold, and valence) and detection of odors are tightly involved with respiration, the subject's respiration pattern modulates and interacts with the experimental paradigm, which presents difficulties for olfactory fMRI data acquisition, post‐processing, and interpretation. Based on our investigation on the interactions of odor presentation and subject's respiration, we propose a respiration‐triggered event‐related olfactory fMRI technique and a data post‐processing method that effectively captures precise onsets of olfactory blood‐oxygen‐level‐dependent (BOLD) signal in the primary olfactory cortex. We compared the olfactory BOLD signals from seventeen normal healthy adults with diverse respiratory patterns and showed that the subjects' respiratory patterns modulated the olfactory stimulation paradigm, which significantly confounded the BOLD signal. The presented experimental technique provides a simple and effective means for generating reliable olfactory fMRI results. Hum Brain Mapp 35:3616–3624, 2014. © 2013 Wiley Periodicals, Inc .     

Neural correlates of evaluative compared with passive tasting

We used functional magnetic resonance imaging to test the hypothesis that the nature of the neural response to taste varies as a function of the task the subject is asked to perform. Subjects received sweet, sour, salty and tasteless solutions passively and while evaluating stimulus presence, pleasantness and identity. Within the insula and overlying operculum the location of maximal response to taste vs. tasteless varied as a function of task; however, the primary taste cortex (anterior dorsal insula/frontal operculum – AIFO), as well as a more ventral region of anterior insula, responded to taste vs. tasteless irrespective of task. Although the response here did not depend upon task, preferential connectivity between AIFO and the amygdala (bilaterally) was observed when subjects tasted passively compared with when they performed a task. This suggests that information transfer between AIFO and the amygdala is maximal during implicit processing of taste. In contrast, a region of the left lateral orbitofrontal cortex (OFC) responded preferentially to taste and to tasteless when subjects evaluated pleasantness, and was preferentially connected to earlier gustatory relays (caudomedial OFC and AIFO) when a taste was present. This suggests that processing in the lateral OFC organizes the retrieval of gustatory information from earlier relays in the service of computing perceived pleasantness. These findings show that neural encoding of taste varies as a function of task beyond that of the initial cortical representation.     

Umami: a delicious flavor formed by convergence of taste and olfactory pathways in the human brain

Umami taste is produced by glutamate acting on a fifth taste system. However, glutamate presented alone as a taste stimulus is not highly pleasant, and does not act synergistically with other tastes (sweet, salt, bitter and sour). We show here that when glutamate is given in combination with a consonant, savory, odour (vegetable), the resulting flavor can be much more pleasant. Moreover, we showed using functional brain imaging with fMRI that the glutamate taste and savory odour combination produced much greater activation of the medial orbitofrontal cortex and pregenual cingulate cortex than the sum of the activations by the taste and olfactory components presented separately. Supralinear effects were much less (and significantly less) evident for sodium chloride and vegetable odour. Further, activations in these brain regions were correlated with the pleasantness and fullness of the flavor, and with the consonance of the taste and olfactory components. Supralinear effects of glutamate taste and savory odour were not found in the insular primary taste cortex. We thus propose that glutamate acts by the nonlinear effects it can produce when combined with a consonant odour in multimodal cortical taste-olfactory convergence regions. We propose the concept that umami can be thought of as a rich and delicious flavor that is produced by a combination of glutamate taste and a consonant savory odour. Glutamate is thus a flavor enhancer because of the way that it can combine supralinearly with consonant odours in cortical areas where the taste and olfactory pathways converge far beyond the receptors.     

Flavor and the Formation of Category-Specific Processing in Olfaction

The perception of flavor occurs when objects, such as food and drink, are placed in the mouth. Although the sensation that ensues depends upon inputs from multiple sensory modalities, due to a combination of oral referral and common sensory qualities (e.g., odors and tastes can both be sweet), it is experienced as a unitary flavor perception. In this paper, it is proposed that neural processing within the somatomotor mouth area of the Rolandic operculum mediates oral referral and causes the neural binding of multimodal inputs to create a flavor percept. It is further proposed that unimodal taste and unimodal smell neurons alter the selectivity of bimodal taste/smell cells only if the binding mechanism in the somatomotor mouth area is active. The encoded flavor object is thus represented by a bounded pattern of response that includes the sculpted bimodal cells as well as the unimodal responses distributed across the insula, operculum, anterior cingulate cortex, and orbitofrontal cortex. Once an odor is encoded in this way, the odor acquires the ability to reactivate this encoded percept, whether experienced orthonasally or retronasally. Finally, it is proposed that one manifestation of this process is the existence of category-specific processing in olfaction.     

The evaluation of brain activity in response to taste stimuli--a pilot study and method for central taste activation as assessed by event-related fMRI

BACKGROUND: Brain pathways contribute to the regulation of appetite behaviors, and advancements in brain imaging offer new opportunities in determining whether disturbances of these pathways play a role in pathological feeding behaviors in humans. We developed a standardized method for the assessment of brain activation in response to taste stimuli. METHODS: Five healthy control women were positioned in a 1.5 T GE magnet resonance (MR) scanner for functional MR imaging (fMRI). They received 1.0 cm3 samples of 1 M glucose solution or artificial saliva (25 mM KCl, 2 mM NaHCO3). Fluid challenges were delivered by a programmable syringe pump (J-Kem Scientific, St. Louis, MO). E-Prime software (Psychology Software Tools Inc., Pittsburgh, PA) coordinated taste stimulation with MR scanning. Data were analyzed using NeuroImaging software (NIS). RESULTS: Healthy women showed increased orbitofrontal cortex activation when glucose was compared to artificial saliva. In addition, mesial and lateral temporal cortical regions contrasted glucose from artificial saliva. CONCLUSIONS: This study demonstrates a design for the systematic study of brain activation after taste stimulation using fMRI and computer controlled stimulus delivery. The results are consistent with previous studies, showing activation in higher order brain centers that are involved in emotional coding of taste experience.     

Functional MRI at 4.7 tesla of the rat brain during electric stimulation of forepaw, hindpaw, or tail in single- and multislice experiments

Stimulation of peripheral nerves activates corresponding regions in sensorimotor cortex. We have applied functional magnetic resonance imaging (fMRI) techniques to monitor activated brain regions by means of measuring changes of blood oxygenation level-dependent contrast during electric stimulation of the forepaw, hindpaw, or tail in rats. During alpha-chloralose anesthesia, artificial respiration, and complete muscle relaxation, stimulations were delivered at 3 Hz via subcutaneous bipolar electrodes with 500-microseconds-current pulses of 0.2-2.0 mA. Single- or multislice gradient echo images were collected during recording sessions consisting of five alternating rest and stimulation periods. Stimulation of the right and left forepaws and hindpaws repeatedly led to robust activation of the contralateral sensorimotor cortex. There was a significant correlation (P < 0.05) between current pulse strength and amount of activation of the sensory cortex during forepaw stimulation. The center of the main cortical representation of the forepaw was situated 3.4 mm lateral to the midline and 5 mm posterior to the rhinal fissure. The main representation of the hindpaw was 2.0 mm lateral to the midline and 6 mm posterior to the rhinal fissure. Tail stimulation gave rise to a strikingly extended bilateral cortical activation, localized along the midline in medial parietal and frontal cortex 4 and 5 mm posterior to the rhinal fissure. In conclusion, the experiments provide evidence that peripheral nerve stimulation induces a fMRI signal in the respective division of the somatosensory cortex in a stimulus-related manner. The marked cortical activation elicited by tail stimulation underlines the key importance of the tail.