Brainstem changes in 5-HT1A receptor availability during migraine attack.

BACKGROUND: Among serotonin receptors, 5-HT(1A) receptors are implicated in the regulation of central serotoninergic tone and could be involved in the abnormal brain 5-HT turnover suspected in migraineurs. The aim of this study was to investigate 5-HT(1A) receptors' availability during migraine attacks. METHODS: Ten patients suffering from odor-triggered migraine attacks and 10 control subjects were investigated using positron emission tomography (PET) and [(18)F]MPPF PET tracer, a selective 5-HT(1A) antagonist. All subjects underwent calibrated olfactory stimulations prior to the PET study. RESULTS: Four patients developed a migraine attack during the PET study. In these patients, statistical parametrical mapping and region of interest analyses showed an increased [(18)F]MPPF binding potential (BP(ND)) in the pontine raphe when compared to headache-free migraineurs and control subjects. This ictal change was confirmed at the individual level in each of the four affected patients. In comparison with the headache-free migraineurs, patients with a migraine attack also showed significantly increased [(18)F]MPPF BP(ND) in the left orbitofrontal cortex, precentral gyrus and temporal pole. No significant change in [(18)F]MPPF BP(ND) was observed between headache-free migraineurs and controls. CONCLUSIONS: Our results emphasize the role of 5HT(1A) receptors in the pontine raphe nuclei during the early stage of migraine attacks.

Subliminal fear priming potentiates negative facial reactions to food pictures in women with anorexia nervosa.

BACKGROUND: To investigate hedonic reactivity and the influence of unconscious emotional processes on the low sensitivity to positive reinforcement of food in anorexia nervosa (AN). METHOD: AN and healthy women were exposed to palatable food pictures just after a subliminal exposure to facial expressions (happy, disgust, fear and neutral faces), either while fasting or after a standardized meal (hunger versus satiety). Both implicit [facial electromyographic (EMG) activity from zygomatic and corrugator muscles, skin conductance, heart rate, and videotaped facial behavior] and explicit (self-reported pleasure and desire) measures of affective processes were recorded. RESULTS: In contrast to healthy women, the AN patients did not display objective and subjective indices of pleasure to food pictures when they were in the hunger states. Pleasure to food cues (liking) was more affected than the desire to eat (wanting) in AN patients. Subliminal 'fear faces' increased corrugator muscle reactivity to food stimuli in fasting AN patients, as compared to controls. CONCLUSIONS: The results suggest that unconscious fear cues increase the negative appraisal of alimentary stimuli in AN patients and thus contribute to decreased energy intake.

Cerebral imaging and olfactory disorders: a review.

Cerebral imaging and olfactory disorders: a review. Olfactory disorders are often misjudged and rarely given due clinical consideration. Nevertheless, they occur in a wide range of neurological disorders, and their evaluation can help in diagnosis. Whereas psychophysical tests have been used to evaluate olfactory dysfunction in numerous diseases, functional brain imaging using olfactory stimuli is an emergent technique and few studies have been published to date. After a reminder of cerebral imaging and analysis techniques and a rapid description of our actual knowledge of olfactory processes in healthy subjects, the current review focuses on cerebral imaging studies performed on patients with neurological disorders and presenting olfactory dysfunction. Neurological disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, migraine, multiple chemical sensitivity and schizophrenia are examined.

Olfactory hypersensitivity in migraineurs: a H(2)(15)O-PET study.

Olfactory hypersensitivity (OHS) may occur during migraine attacks and seems to be very specific to this form of headache. OHS is also observed during migraine-free periods and is associated with the presence of odour-triggered attacks. Yet the pathophysiology of OHS remains unknown. The aim of our study was to evaluate olfactory processing in migraineurs with OHS and to investigate whether regional cerebral blood flow (rCBF) associated with olfactory stimulation is modified in these patients compared with controls. Eleven migraineurs with OHS and 12 controls participated in a H(2)(15)O-positron emission tomography study, including three scans in which odours were delivered and three scans where only odourless air was delivered. rCBF during olfactory condition was compared with that for the odourless baseline condition. Between-group analyses were performed using voxel-based and region-of-interest analyses. During both olfactory and non-olfactory conditions, we observed higher rCBF in the left piriform cortex and antero-superior temporal gyrus in migraineurs compared with controls. During odour stimulation, migraineurs also showed significantly higher activation than controls in the left temporal pole and significantly lower activation in the frontal (left inferior as well as left and right middle frontal gyri) and temporo-parietal (left and right angular, and right posterior superior temporal gyri) regions, posterior cingulate gyrus and right locus coeruleus. These results could reflect a particular role of both the piriform cortex and antero-superior temporal gyrus in OHS and odour-triggered migraine. Whether these rCBF changes are the cause or a consequence of odour-triggered migraines and interictal OHS remains unknown. Further comparisons between migraineurs with and without OHS are warranted to address this issue. The abnormal cerebral activation patterns during olfactory stimulation might reflect altered cerebrovascular response to olfactory stimulation due to the migraine disease, or an abnormal top-down regulation process related to OHS.

[Olfaction and neurological diseases: a review of the literature]. / Olfaction et pathologies neurologiques: revue de la littérature.

Olfactory disorders are often misjudged and rarely rated in the clinical setting. They are nevertheless described in a wide range of neurological disorders, and their evaluation can be useful for diagnosis. Usually irreversible olfactory dysfunction is a well-known complication after head trauma. Severe changes in olfactory tests are observed in Parkinson's disease. Dysfunction is present at disease onset and evidenced with all behavioral tests. Regarding other parkinsonian syndromes, olfactory performances are severely impaired in Lewy body disease, less pronounced in multiple system atrophy and usually preserved in corticobasal degeneration. Olfactory deficits are an early feature in Alzheimer's disease and worsen with disease progression. Rarely reported by patients, they must be searched for with olfactory tests. Though epilepsy is mainly known for its olfactory hallucinatory disorders, alterations of olfactory abilities are also described, especially in mesial temporal epilepsy. Disorders of olfactory perception are finally reported in patients with multiple sclerosis and migraine. After a reminder of anatomical data on the olfactory system, and the different methods of testing used to rate olfactory performances, the current review focuses on the type of olfactory dysfunction and damaged brain areas of the olfactory system encountered in the main neurological diseases.

Stereology: a method for analyzing images.

This review deals with notions of shape, sizes, numbers, densities and orientation in space, all basic concepts in stereology. With the initiation by Delesse in 1847, but mainly since the beginning of the XXth century, many stereological methods have been published allowing us to relate two-dimensional measurements easily obtainable on flat histological images with three-dimensional characteristics of the structure analysed. Looking at these methods, the neurobiologist, generally impermeable to concepts of sampling, statistical bias, efficiency, cost of effort and distribution-free, is discountenanced and continues old laboratory usages and customs. Furthermore, for the last ten years, the advent of a plethora of new powerful tools, considered as assumption-free and more efficient than the previous ones, increase the risk proportionately the disarray of the potential user. The purpose of this review is to present synthetically all traditional and actual aspects of stereology in order to guide the reader in the labyrinth of this speciality. The necessarily short exposition is compensated by many references to which the beginner or the initiated can refer.

Emotional responses to pleasant and unpleasant olfactory, visual, and auditory stimuli: a positron emission tomography study.

Neural correlates of responses to emotionally valenced olfactory, visual, and auditory stimuli were examined using positron emission tomography. Twelve volunteers were scanned using the water bolus method. For each sensory modality, regional cerebral blood flow (rCBF) during presentation of both pleasant and unpleasant stimuli was compared with that measured during presentation of neutral stimuli. During the emotionally valenced conditions, subjects performed forced-choice pleasant and unpleasant judgments. During the neutral conditions, subjects were asked to select at random one of a two key-press buttons. All stimulations were synchronized with inspiration, using an airflow olfactometer, to present the same number of stimuli for each sensory modality. A no-stimulation control condition was also performed in which no stimulus was presented. For all three sensory modalities, emotionally valenced stimuli led to increased rCBF in the orbitofrontal cortex, the temporal pole, and the superior frontal gyrus, in the left hemisphere. Emotionally valenced olfactory and visual but not auditory stimuli produced additional rCBF increases in the hypothalamus and the subcallosal gyrus. Only emotionally valenced olfactory stimuli induced bilateral rCBF increases in the amygdala. These findings suggest that pleasant and unpleasant emotional judgments recruit the same core network in the left hemisphere, regardless of the sensory modality. This core network is activated in addition to a number of circuits that are specific to individual sensory modalities. Finally, the data suggest a superior potency of emotionally valenced olfactory over visual and auditory stimuli in activating the amygdala.

5-Hydroxytryptamine action in the rat olfactory bulb: in vitro electrophysiological patch-clamp recordings of juxtaglomerular and mitral cells.

The olfactory bulb, first relay of olfactory pathways, is densely innervated by serotoninergic centrifugal fibers originating from the raphe nuclei. Although serotonin innervation was reported to be involved in olfactory learning in mammals, the action of this neurotransmitter on its putative cellular targets has been never described through unitary recordings. This lack of data initiated the present study where the effects of 5HT on juxtaglomerular and mitral cells are analyzed using whole-cell recordings on olfactory bulb slices. Serotonin depolarizes 34% of 525 JG cells. A multivariate statistical analysis of juxtaglomerular cells characteristics shows that the serotonin responsive cell group can be individualized regarding their tonic discharge-mode in response to a direct current injection, their lower expression of hyperpolarization-activated cation current and their low membrane capacities. The use of ion channel blockers and ramp voltage protocol indicate that serotoninergic depolarization of juxtaglomerular cells may be due to a nonselective cation current with a reversal potential of -44 mV. Pharmacological tests with serotonin receptor antagonists and agonists reveal that 5HT action on juxtaglomerular cells would be mainly mediated by 5HT2C receptors. In mitral cells, serotonin acts on 49.1% of the 242 tested cells, inducing two types of responses. A first subset of mitral cells (26.8%, n=65) were hyperpolarized by serotonin. This response would be indirect and mediated by action of GABA on GABAA receptors since it was antagonized by bicuculline. The involved GABAergic neurons are hypothesized to be juxtaglomerular and granular cells, on which serotonin would act mainly via 5HT2C and via 5HT2A receptors respectively. The second subset of mitral cells (22.3%, n=54) were directly depolarized by serotonin acting through 5HT2A receptors. Our data on serotonin action on juxtaglomerular cells and mitral cells reveal a part of functional mechanisms whereby serotonin can act on olfactory bulb network. This is expected to enrich the understanding of its determining role in olfactory learning.

A stimulation method using odors suitable for PET and fMRI studies with recording of physiological and behavioral signals.

A design for a semi-automatic olfactometric system is described for PET and fMRI experiments. The olfactometer presents several advantages because it enables the use of an 'infinite' number of odorants and the synchronization of stimuli with breathing. These advantages mean that the subject is recorded while breathing normally during olfactory judgment tasks. In addition, the design includes a system for recording the behavioral (rating scale) and physiological (breathing, electrodermal reaction (ED), plethysmography (PL)) signals given by the subject. Both systems present the advantage of being compatible with fMRI magnetic fields since no ferrous material is used in the Faraday cage and signals are transmitted via an optical transmission interface to an acquisition system.

Morphometric modifications associated with early sensory experience in the rat olfactory bulb: I. Volumetric study of the bulbar layers.

Current models of sensory coding in the olfactory bulb are based on the notion of topographical specificity in the processing of stimuli. Part of this hypothesis comes from studies of patterns of radiolabelled 2-deoxyglucose uptake, and local morphometric variations of the mitral cell size observed following prolonged exposure to an odor. The present study explored the possibility that exposing young rats to a long-term stimulation with an odor would induce spatially distributed volumetric variations of the bulbar layers. Three groups of 5 rats have been studied: (1) stimulated with ethyl acetoacetate from birth to 1 month of age, (2) unilaterally deprived following early cauterization of one nostril, and (3) normal animals of same age. Fourteen frontal histological sections uniformly distributed along the rostrocaudal axis of the olfactory bulb were used for this study. Areas of the bulbar layers were measured with the aid of an image analyser and the volume of the corresponding layers deduced by computation. Following complete sensory deprivation, the volume of all bulbar layers, except the periventricular core, was homogeneously reduced along the rostrocaudal axis of the olfactory bulb. Following long-term stimulation with ethyl acetoacetate, volume reduction was significantly higher in anterior and middle regions than in the posterior part of the olfactory bulb. It is assumed that neuronal pathways activated by ethyl acetoacetate stimulation are mainly located in the posterior part of the olfactory bulb. Functional interpretations of these results are discussed with respect to the spatial dimension in olfactory coding.

Morphometric modifications associated with early sensory experience in the rat olfactory bulb: II. Stereological study of the population of olfactory glomeruli.

The present study explores the local variations of size and number of olfactory glomeruli induced by the exposure of young rats to long-term stimulation with a single odor. Three groups of 5 rats were used that were either: (1) stimulated with ethyl acetoacetate from birth to 1 month of age, (2) unilaterally deprived following early occlusion of one nare, or (3) normal animals of the same age. Areas and coordinates of all glomerular profiles were measured in 14 coronal sections uniformly distributed along the rostrocaudal axis of the olfactory bulb. A distribution-free stereological method was applied to compute the size and number of glomeruli either along the bulbar rostrocaudal extent or in the bulbar coronal plane. Following complete sensory deprivation or long-term stimulation with ethyl acetoacetate, the mean diameter of glomeruli was significantly reduced everywhere, except in the ventrolateral and ventromedial regions of the posterior olfactory bulb in rats reared with a single odor. In both of these areas, the number of glomeruli was either significantly increased following long duration exposure or significantly reduced following unilateral deprivation. Thus these results show that selective modifications of the olfactory environment during postnatal maturation induce morphometric variations in specific areas of the glomerular layer. These data are discussed with respect to the concept of the topographical coding of odor quality at the level of the glomeruli and plasticity of the olfactory system during postnatal development.

Morphometric study of the glomerular population in the mouse olfactory bulb: numerical density and size distribution along the rostrocaudal axis.

A morphometric study of the glomerular population in the olfactory bulb of the mouse has been carried out by using stereological methods. On the basis of the assumption that the glomerular population is a polydispersed system of spheres, glomerular profile distributions obtained from profile measurements were subjected to a mathematical unfolding procedure to obtain the actual glomerular size distribution. We used a distribution-free method to account for the combined effects of overprojection due to section thickness and truncation (two missing profile mechanisms). Results proved better than those obtained directly from profile measurements without stereological analysis. Several new findings were obtained. First, significant variations of the glomerulus sizes were found along the rostrocaudal axis. The glomeruli are larger in the middle region of the olfactory bulb, whereas their numerical density decreases in the same region. Moreover, the profile density is homogeneous along the rostrocaudal axis. In other words, the relative surface occupied by the periglomerular cells in the glomerular layer is invariant. As a consequence, it may be concluded that the variations in size and numerical density are inversely correlated. Thus, since the glomeruli are larger in the middle region, their number per unit volume is logically smaller in this same area. Finally, the computerization of all these data has led us to estimate the number of glomeruli (1,810 +/- 41) in the olfactory bulb of the mouse. In order to get a comparative idea of their advantages and disadvantages, other standard stereological methods were used in the present study to determine this number. Functional interpretations of the variations of the size and numerical density along the rostrocaudal axis of the olfactory bulb are discussed with respect to ontogenetic and morphofunctional data obtained elsewhere.

An olfactometric cage suitable for short duration stimulations of unrestrained small animals.

An olfactometric cage was constructed in order to perform experiments on olfactory perception in unrestrained rats. Olfactory stimuli were measured with a flame ionization detector (FID). A cylindrical cage was divided into 2 compartments, an upper expansion chamber (B) and a lower stimulation chamber (A) with a perforated metal plate. Before entering B, the odours were diluted with purified air in a mixing chamber. The odorized air was then delivered to a small chamber on top of the cage and distributed by a thin slit surrounding B. A homogeneous distribution of holes in the metal plate provided a laminar flow characterized by a parabolic front. This flow was suitable for long duration exposures of animals of odours. However, for short stimulations (10 sec), variations of amplitude and delay were observed in the horizontal plane of the olfactometric cage. To flatten this parabolic front a new distribution of holes was calculated. A partially turbulent air flow was obtained in A. creating some fluctuations in the time-course of stimulations. However, the amplitude and the delay of the signals were reasonably constant in all parts of the cage floor. It is possible to use this apparatus for other purposes than those originally intended, e.g. behavioural studies.

Lesions of the olfactory pathways affecting neophobia and learned aversion differentially.

The contribution of ascending olfactory pathways in neophobia and learned aversion to the same food was investigated in male rats bearing lesions of both olfactory peduncles, or one olfactory peduncle and the opposite lateral olfactory tract or anterior limb of the anterior commissure. The animals were fed on usual stock diet (S) offered as a choice with novel vanilla food (V) on test days: during neophobia (N), then before and after aversive conditioning (Aa, At). Daily food intake was measured, and the preference was expressed as V/(V + S). Experiment 1 included a neophobia test, before aversive conditioning (3 mEq/kg LiCl, i.p.). In Experiment 2, aversion only was studied (0.9 mEq/kg). In the neophobia test, the preference ratio was 7% in unoperated controls, and 43-52% in the 3 lesioned groups. The same controls had preference ratios equal to 64% and 22%, before and after aversive learning. Similar drops were observed for any lesioned group in Expt. 1. The decrease was less obvious, although significant, in rats of Expt. 2 with asymmetric lesions; those with both olfactory peduncles cut through maintained the same preference ratio (48%) before and after LiCl treatment. The data are interpreted assuming that: (1) lateral olfactory tract and anterior commissure both contribute to information processing in neophobia and aversion; (2) olfactory cues subserve neophobia prepotently; and (3) one cannot account for the sensory determinism of neophobia and aversion calling for a single mechanism.

Effect of local 6-OHDA and 5,6-DHT injections into the rat olfactory bulb on neophobia and learned aversion to a novel food.

This study investigated the role of the noradrenergic (NA) and the serotoninergic (5-HT) centrifugal fibers on the olfactory bulb (OB). For this purpose, behavioral responses largely depending on olfactory information were measured in rats after discrete bilateral injections into the OB with either 6-hydroxydopamine (6-OHDA) or 5,6-dihydroxytryptamine (5,6-DHT) solutions. The evaluation of the level of endogenous 5-HT in the OB was measured by high performance liquid chromatography. The NA fibers appeared lesioned after the 6-OHDA treatment only, whereas the 5-HT level was significantly reduced in 5,6-DHT treated bulbs only (34% of the control value). The animals were fed on usual stock-diet (S) and had the choice between S and a novel food (V) on neophobia test, and again on the following day after aversive conditioning (0.9 mEq/kg LiCl i.p.). The behavioral results showed that the drug administrations were followed by a significant effect in one case only: the intake of V was significantly enhanced in 6-OHDA treated animals during neophobia test. Thus, neophobia was reduced by 6-OHDA. The data are interpreted as follows: (1) the lesion of the bulbar NA or 5-HT fibers does not induce total anosmia; and (2) the NA innervation of the OB could play a role in the modulation of behavioral responses to biological odorants.

A comparative study of 2-deoxyglucose patterns of glomerular activation in the olfactory bulbs of C57 BL/6J and AKR/J mice.

We have studied the patterns of glomerular activation evoked in the olfactory bulbs of C57BL/6J and AKR/J mice by olfactory stimulations with amyl acetate or isovaleric acid. Patterns of glomerular activation were obtained with the 2-deoxyglucose method and subsequently compared using a computer-assisted image analysis. The mice of both inbred strains stimulated with amyl acetate or isovaleric acid were characterized by areas of high 2-deoxyglucose uptake in their glomerular layer. Statistical comparisons of the patterns demonstrated that they were odor-specific. Although C57BL/6J mice have been found to be specifically anosmic to isovaleric acid, our findings indicate that their olfactory system is topographically activated by olfactory stimulations with this odorant. However, patterns of glomerular activity evoked by isovaleric acid stimulations are not similar in the two studied strains.

Specificity of spatial patterns of glomerular activation in the mouse olfactory bulb: computer-assisted image analysis of 2-deoxyglucose autoradiograms.

We have developed a computer-assisted method for analyzing the 2-deoxyglucose (2-DG) autoradiograms of mice olfactory bulbs. The purpose of the study was to numerize the maps of glomerular activation in order to achieve a statistical comparison of the glomerular patterns evoked by different stimuli. The spatial distribution of glomerular activation was displayed on unfolded representations of the glomerular layer which were built up using glomerular optical densities (OD) measured systematically within 13 sections per bulb. Each bulbar sample was converted into an 'OD profile'. A matrix composed of 18 OD profiles was submitted to a principal component analysis. The first factor which accounted for 28% of the variance separated unambiguously two clusters corresponding to the bulbs issued from animals stimulated with amylacetate and isovaleric acid, respectively. The second and third factors which accounted for 14% and 12% of the variance segregated the control group (animals exposed to pure air) from the odor-stimulated ones. It was demonstrated that the cluster separation was actually due to the specific spatial distribution of the most-labelled glomeruli. A particular attention was paid to the well-delineated glomerular activation evoked by isovaleric acid. The results demonstrate the specificity and reliability of the glomerular 2-DG patterns. The method should be useful for further comparisons of patterns elicited by larger sets of odorant compounds.

A re-estimation of the number of glomeruli and mitral cells in the olfactory bulb of rabbit.

Although morphological characteristics of mammalian olfactory bulb (OB) are well documented in rodents (rat, mice), only one study has been performed in rabbit, which is also commonly used in olfactory research. The study carried out by Allison and Warwick in 1949 led to surprising results still quoted in recent literature. The present study re-examined this question in young rabbit OB, while it was also conducted with rat as control. In five animals of each species, areas and coordinates of glomeruli and mitral profiles were measured in 10 coronal sections uniformly distributed along the rostrocaudal axis of the OB, and a distribution-free stereological method was used to compute values along this axis. For glomeruli, the estimated number was 4200 in rat and 6300 in rabbit. While this estimation matched with those found in literature in rat, it strikingly differed from the Allison and Warwick's value of 1900 only. For mitral cell number, we found 59,600 while the preceding study found 45,000 only. In contrast to the number of glomeruli, the mitral cell number in rat and rabbit were very close. Indeed, results showed 56, 200 in rat. The results suggest that the numbers of olfactory glomeruli and mitral cells has been previously underestimated in rabbit, and that the number of glomeruli changes as a function of species. In addition, both the results of the present study and reports in the literature suggest the number of mitral cells to be rather similar in mouse, rat and rabbit. As a consequence, the glomerular/mitral cell ratio is likely to varied in a wide range across species.

Olfactory bulb responsiveness to an aversive or novel food odor in ;the unrestrained rat.

The mechanisms subserving neophobia and learned aversion have been investigated by recording multiunit olfactory bulb discharges either in hungry rats following food deprivation or in satiated rats. Under the two conditions, rats were stimulated with the smell of their familiar maintenance diet or that of a novel food or of control food-unrelated odor. Responses to the odor of the novel food were tested, following a pairing of the first or the second intake of that food with a LiCl injection, or following its first intake paired with a NaCl control injection. All rats exhibited enhanced level of discharges when they were stimulated in the hungry state with the smell of the familiar food and not when stimulated with the non-alimentary control odor. The hunger to satiety modulation of olfactory bulb discharges, also exhibited in rats tested with the smell of the novel food, previously paired with NaCl, was absent after a LiCl-induced taste aversion to this odor. The small, although significant, modulation observed when the conditioning of aversion occurred with the less novel food is consistent with the view that learned safety prevails upon learned harmfulness. Results are discussed in terms of relations of olfactory bulb electrical responses to odors with food palatability, neophobia and learned aversion.