Decision-making in a changing world: a study in autism spectrum disorders.

To learn to deal with the unexpected is essential to adaptation to a social, therefore often unpredictable environment. Fourteen adults with autism spectrum disorders (ASD) and 15 controls underwent a decision-making task aimed at investigating the influence of either a social or a non-social environment, and its interaction with either a stable (with constant probabilities) or an unstable (with changing probabilities) context on their performance. Participants with ASD presented with difficulties in accessing underlying statistical rules in an unstable context, a deficit especially enhanced in the social environment. These results point out that the difficulties people with ASD encounter in their social life might be caused by impaired social cues processing and by the unpredictability associated with the social world.

Brain correlates of automatic visual change detection.

A number of studies support the presence of visual automatic detection of change, but little is known about the brain generators involved in such processing and about the modulation of brain activity according to the salience of the stimulus. The study presented here was designed to locate the brain activity elicited by unattended visual deviant and novel stimuli using fMRI. Seventeen adult participants were presented with a passive visual oddball sequence while performing a concurrent visual task. Variations in BOLD signal were observed in the modality-specific sensory cortex, but also in non-specific areas involved in preattentional processing of changing events. A degree-of-deviance effect was observed, since novel stimuli elicited more activity in the sensory occipital regions and at the medial frontal site than small changes. These findings could be compared to those obtained in the auditory modality and might suggest a "general" change detection process operating in several sensory modalities.

Schizophrenia and frontal cortex: where does it fail?

Schizophrenia is characterized by cognitive, social, and emotional impairments and by psychotic symptoms. Neuroimaging studies have reported abnormalities within the prefrontal cortex and it has been hypothesized that schizophrenia results from poor or miswired anatomical/functional connections. We have compared the functional connectivity within the frontal cortex in control and schizophrenic subjects during the realization of a Continuous Performance Task. The connectivity pattern within the frontal cortex was uncovered by the analysis of the correlation matrix computed from the fMRI time series in frontal areas for 14 schizophrenic patients and 14 control subjects. In control subjects, the right dorsolateral prefrontal cortex (DLFCr) activity correlated i) positively with the left dorsolateral prefrontal cortex and the posterior part of the supplementary motor area, ii) negatively with the medial and anterior/inferior part of the frontal cortex. In the schizophrenic group, these relations were abolished or strongly lowered. The negative relation between the DLFCr and the medial frontal cortex has been proposed to play a key role in setting a harmonious balance between the direction of attention to the external world and the expression of the individual believes and self-referential activities, and therefore, the impaired relation of right DLFCr with other frontal areas could explain a distorted perception of external world in relation with internal motivations.

Generalized least-squares method applied to fMRI time series with empirically determined correlation matrix.

Functional magnetic resonance imaging (fMRI) time series analysis and statistical inferences about the effect of a cognitive task on the regional cerebral blood flow (rCBF) are largely based on the linear model. However, this method requires that the error vector is a gaussian variable with an identity correlation matrix. When this assumption cannot be accepted, statistical inferences can be made using generalized least squares. In this case, knowledge of the covariance matrix of the error vector is needed. In the present report, we propose a method that needs stationarity of the autocorrelation function but is more flexible than autoregressive model of order p (AR(p)) models because it is not necessary to predefine a relation between coefficients of the correlation matrix. We tested this method on sets of simulated data (with presence of an effect of interest or not) representing a time series with a monotonically decreasing autocorrelation function. This time series mimicked an experiment using a random event-related design that does not create correlation between scans. The autocorrelation function is empirically determined and used to reconstitute the correlation matrix as the toeplitz matrix built from the autocorrelation function. When applied to simulated time series with no effect of interest, this method allows the determination of F values corresponding to the accurate false positive level. Moreover, when applied to time series with an effect of interest, this method gives a density function of F values which allows the rejection of the null hypothesis. This method provides a flexible but interpretable time domain noise model.

A neural network elicited by parametric manipulation of the attention load.

We used a parametric experimental design to identify the rCBF variations related to a continuous variation of the attention load. The experiment involved goal-directed visual tasks. The length of time during which the subject's attention was engaged toward the external stimulus was taken as the factor of interest. The neural network revealed areas that positively (left cerebellum, bilateral MT/V5 complex and superior parietal lobule, right inferior temporal lobe and dorsolateral prefrontal cortex) or negatively (precuneus, anterior cingulate and medial superior frontal cortex) correlate with the attention load. Results demonstrate that the activity of these areas varies continuously as a function of the variation in the attention load.

How the brain perceives causality: an event-related fMRI study.

Detection of the causal relationships between events is fundamental for understanding the world around us. We report an event-related fMRI study designed to investigate how the human brain processes the perception of mechanical causality. Subjects were presented with mechanically causal events (in which a ball collides with and causes movement of another ball) and non-causal events (in which no contact is made between the balls). There was a significantly higher level of activation of V5/MT/MST bilaterally, the superior temporal sulcus bilaterally and the left intraparietal sulcus to causal relative to non-causal events. Directing attention to the causal nature of the stimuli had no significant effect on the neural processing of the causal events. These results support theories of causality suggesting that the perception of elementary mechanical causality events is automatically processed by the visual system.

Does perception of biological motion rely on specific brain regions?

Perception of biological motions plays a major adaptive role in identifying, interpreting, and predicting the actions of others. It may therefore be hypothesized that the perception of biological motions is subserved by a specific neural network. Here we used fMRI to verify this hypothesis. In a group of 10 healthy volunteers, we explored the hemodynamic responses to seven types of visual motion displays: drifting random dots, random dot cube, random dot cube with masking elements, upright point-light walker, inverted point-light walker, upright point-light walker display with masking elements, and inverted point-light walker display with masking elements. A gradient in activation was observed in the occipitotemporal junction. The responses to rigid motion were localized posteriorly to those responses elicited by nonrigid motions. Our results demonstrate that in addition to the posterior portion of superior temporal sulcus, the left intraparietal cortex is involved in the perception of nonrigid biological motions.

New aspects of motion perception: selective neural encoding of apparent human movements.

Perception of apparent motion operates somewhat differently for objects and human figures. Depending on the interstimulus interval, the latter d may give rise to either perception of a direct path (i.e. biologically impossible) or indirect path (i.e. biologically possible). Here, PET was used to investigate whether a change in brain activity accompanies this perceptual shift. We found neural encoding of apparent motion to be a function of the intrinsic properties of the stimulus presented (object vs human) as well as the kind of human movement path perceived (biomechanically possible vs impossible). Motor and parietal cortex were only involved for possible motion which suggests that these regions are selectively activated to process actions which conform to the capabilities of the observer.

Susceptibility of differentiated thyrocytes in primary culture to undergo apoptosis after exposure to hydrogen peroxide: relation with the level of expression of apoptosis regulatory proteins, Bcl-2 and Bax.

Thyrocytes, that generate and use hydrogen peroxide (H2O2) to synthesize thyroid hormones, undergo apoptosis, as do most cell types, when exposed in vitro to H2O2. We have studied 1) the kinetics and the amplitude of the apoptotic response to H2O2 and 2) the relationship between the extent of the apoptosis-inducing effect of H2O2, the H2O2 degradation activity, and the level of expression of apoptosis regulatory proteins, Bcl-2 and Bax, in pig thyrocytes in primary culture. Cells were seeded at high density to obtain confluent monolayers and were cultured in the presence of TSH to maintain the expression of differentiation. H2O2 (10-300 microM) induced the appearance of cells with fragmented DNA (terminal transferase deoxy-UTP-fluorescein isothiocyanate nick end labeling-positive cells) at a maximum of 3-4 h after H2O2 addition and then the detachment of apoptotic cells from the cell monolayer. The proportion of detached cells increased with H2O2 concentration and amounted to up to 30% of the initial cell number after 24 h. The transient effect of H2O2 was related to its rapid degradation by cells and culture medium components (rate constant, approximately 0.1 min(-1)). Iterative additions of H2O2 produced cumulative apoptotic waves. The amplitude of the apoptotic response of thyrocytes to H2O2 progressively increased with the time of culture, up to 4-fold from days 1-8. This was not related to a change in the capacity of thyrocytes to degrade H2O2. During the same period of culture, the Bcl-2 cell content progressively decreased, whereas that of Bax concomitantly increased; thus, the Bcl-2/Bax ratio varied from about 6 on day 1 to 0.5 on day 10. These data show that the susceptibility of thyrocytes to undergo apoptosis increases with the time of culture and that the pronounced changes in the apoptotic status ofthyrocytes might be linked to coordinate modifications of the level of expression of pro- and antiapoptotic regulatory proteins.

H2O2 induces apoptosis of pig thyrocytes in culture.

Apoptosis might be involved in the reduction of the thyroid cell population in physiopathological situations such as goitre involution and autoimmune deleterious processes. Up to now, little attention has been paid to the apoptotic phenomenon in the normal thyroid gland the specialized metabolism of which is expected to generate reactive oxygen species. Indeed, thyroid cells have the capacity to synthesize H2O2. In this study, we have analyzed the capacity of H2O2 to trigger apoptosis of pig thyrocytes in culture to try to determine whether thyrocytes exhibit a particular resistance to apoptosis induced by an oxidative stress. We show that exposure of thyrocytes cultured as monolayers to exogenous H2O2 induced cell death with characteristics of apoptosis. The effect of H2O2 was concentration-dependent; apoptotic cells were already observed after exposure to 50 micro M H2O2. At high concentrations (millimolar range), H2O2 exerted toxic effects leading to rapid cell disruption. Within the first hour after the onset of exposure to 50-300 micro M H2O2, early signs of apoptosis, i.e. DNA fragmentation, appeared in a low (0.1-1%) but definite fraction of thyrocytes. The proportion of adherent cells exhibiting DNA fragmentation remained fairly constant after 6, 15 and 24 h. During the 24-h period, an increasing number of cells detached from the culture dish and up to 30-40% of cells in suspension displayed apoptotic features. The fraction of cells that lost contact with the culture dish amounted to up to 25% 24 h after addition of 300 micro M H2O2. In conclusion, as reported for other cell types, low H2O2 concentrations are capable of triggering apoptosis in thyrocytes cultured as monolayers. Thyrocytes that undergo apoptosis secondarily lose contact with neighbour cells and the substratum; cell detachment from the monolayer probably happens within 1-2 h after initiation of DNA fragmentation. Our data show that the apoptotic commitment can take place many hours after initiation of the oxidative stress.

Calcium is transported into the lumen of pig thyroid follicles by fluid phase basolateral to apical transcytosis.

The lumen of thyroid follicles contains a high concentration of thyroglobulin, the thyroid prohormone and a high concentration of calcium (Ca2+). As thyroglobulin binds Ca2+, intraluminal Ca2+ is expected to be in free and protein-bound forms. In the present work, we have investigated the mechanism(s) by which Ca2+ could enter the lumen of thyroid follicles. 45Ca2+ uptake studies were carried out on reconstituted pig thyroid follicles (RTF) and pig thyroid cell monolayers (TCM) in primary culture, representing experimental systems with two compartments (cells + lumina) and one compartment, respectively. 45Ca2+ accumulation in RTF was rapid during the first hour of incubation and then slowly increased. Analysis of the uptake data with a "two compartments" model gave two kinetic constant values: k = 1.71 +/- 0.28 hr(-1) and k(-2) = 0.20 +/- 0.05 hr(-1) (n = 10). The slow uptake process accounted for 20-50% of the total RTF-associated Ca2+ after 24 hr. 45Ca2+ uptake by TCM was rapid and reached a stable level within 1-2 hr. Experimental data fitted with a "single compartment" model and gave a k(-1) value of 1.64 +/- 0.15 hr(-1) (n = 10) which was not statistically different from the k(-1) obtained for 45Ca2+ uptake by RTF. We then compared the kinetics of 45Ca2+ uptake by RTF with the kinetics of transport of fluid phase markers: [14C]-sucrose and Lucifer Yellow from the medium to the lumen of RTF. [14C]-sucrose and Lucifer Yellow uptakes by RTF appeared as slow processes compatible with the entry in a single compartment with k values of 0.32 +/- 0.06 hr(-1) (n = 3) and 0.23 +/- 0.015 hr(-1) (n = 3), respectively. These values were not significantly different from the k(-2) value obtained for 45Ca2+ uptake by RTF. These data suggest that thyroid follicles would possess two independent Ca2+ compartments: cells and lumen, and that the entry of Ca2+ into the lumen of follicles probably could take place by fluid phase basolateral to apical transcytosis.

Relationships between pathological diagnosis and clinical parameters in acromegaly.

From our series of 185 somatotropic adenomas with acromegaly, we found that sparsely granulated adenomas were more frequent (56%) than densely granulated ones. Immunocytochemistry detected growth hormone (GH) plurihormonal adenomas in 68% of patients. GH-alpha-subunit (alpha SU) and GH-alpha SU-prolactin (PRL) were more frequent (38%) than GH monohormonal adenomas (32%). The colocalization of GH and alpha SU in the same cell was obvious in many tumors. In contrast, colocalization of GH and PRL was demonstrated in only 25% of GH-PRL adenomas. The relationships between age, sex, tumor size, GH and PRL plasma levels, granularity, and percentage of GH-, alpha SU-, and PRL-immunoreactive cells were established in 105 acromegalic patients by three statistical methods, mainly by a principal component analysis. Correlations were found between the percentage of alpha SU- and GH-immunoreactive cells, and between densely granulated character and the percentage of GH-immunoreactive cells. Tumor size was not correlated with alpha SU, but was positively correlated with PRL plasma levels. Patients' age and percentage of GH-immunoreactive cells were inversely related to tumor size. Plurisecretion and sparsely granulated aspect are not related to age and tumor size.

Cell-to-cell communication in the anterior pituitary: evidence for gap junction-mediated exchanges between endocrine cells and folliculostellate cells.

The ability of rat anterior pituitary cells to communicate through gap junctions (GJ) was studied using a fluorescent molecule, Lucifer Yellow (LY), which freely passes through GJ channels. The probe was introduced into the cell cytoplasm by using either the cut-end loading method on intact tissue, or cell microinjection on cultured cells. The identification of communicating cells was performed by immunofluorescence labeling of specific hormones in endocrine cells and of S100 protein in folliculostellate (FS) cells. Rat anterior pituitary cells in their physiological organization, i.e. in the intact tissue, exhibited a high level of coupling through GJ. LY-labeled cells were found up to 300-microns apart from its site of introduction. The communicating cells were primarily PRL cells, GH cells, and FS cells. Only a few LH, TSH, and ACTH cells were labeled with LY. Anterior pituitary cells, isolated from the rat tissue by mild protease treatment and cultured for 3 days, reestablished functional GJ as demonstrated by microinjection of LY into individual cells. By immunolabeling of specific hormones and/or S100 protein, we found a GJ coupling between FS cells, and between FS cells and endocrine cells, including PRL cells. The communication between FS cells was by far the most frequent. In conclusion, we demonstrate the presence of functional GJ between anterior pituitary cells of the same type and between anterior pituitary cells having distinct differentiated functions.

Quantitative evaluation of the distribution of cell density. Application to human pituitary corticotroph cells.

OBJECTIVE: An image analysis program was developed to analyze cell density in a whole organ. The method was applied to the evaluation of adrenocorticotrophic hormone (ACTH-storing cells in normal human anterior pituitary in order to help the pathologist to evaluate hyperplasia on small surgical biopsies. STUDY DESIGN: ACTH cells were labeled using the immunoperoxidase technique. Data were systematically acquired on a composite image that made possible two-dimensional cell pattern representation in sequential sections of the organ and measurements of cell density in selected histologic parts. RESULTS: ACTH-storing cells were located more in the anterior than posterior part and more in the lower than in the upper pituitary part. The probability of observing high-density value even in normal cases was precisely evaluated zone by zone. Moreover, a model was proposed to evaluate mean cell density from small surgical pieces. CONCLUSION: The same methodology could be applied to other endocrine cell types in the thyroid, pancreas or intestinal tract when hyperplasia is considered the cause of certain clinical syndromes.

Mapping of corticotropic cells in the normal human pituitary.

We accomplished the first mapping of corticotropic cells in the whole human adult pituitary. Corticotropic cells were identified by immunocytochemistry (ICC) and quantified by image analysis on 12 pituitaries obtained from people who had died suddenly. An overall view of each pituitary was given by 15-21 sections (mean 18 sections) at 300-micron intervals on six slides. Each section was systematically treated by indirect immunoperoxidase using an anti-ACTH[17-39] polyclonal antiserum. All the measures were done with a x 6.3 objective lens, each field (0. 5 mm2) being considered as the unit area. The mean pituitary density (surface of labeled cells/total surface) of corticotropic cells (9.5 +/- 3.0% per 0. 5 mm2) is significantly higher in men (11.5 +/- 5.1%) than in women (7.0 +/- 1.3%). This difference is due to an inverse relationship between the corticotropic cell density and the weight of the pituitary, which is higher in women than in men. The mean diameter of corticotropic cells is 14.9 micron and their total number per pituitary is approximately 10(7) cells. We confirmed that the spatial distribution of corticotropic cells is nonuniform: they are mainly distributed in the anteromedian part of the anterior lobe. In addition, our results demonstrated that the inferior part of the pituitary contained three times more corticotropic cells than the superior part (mean density 18.0% vs 6.0%) and the anterior part twice as many as the posterior part (mean density 12.3% vs 6.8%). On the horizontal plane, the pituitary was divided into eight zones, in which the mean of area was 2.5-21.0%. The maximal cell density may reach 40-60%. The use of this map should help the pathologist to recognize if there is corticotropic hyperplasia in a small pituitary fragment surgically removed from a patient with Cushing's disease. On the basis of this study, we put forward some criteria for diagnosing corticotropic hyperplasia.

Gap junctions and cell polarity: connexin32 and connexin43 expressed in polarized thyroid epithelial cells assemble into separate gap junctions, which are located in distinct regions of the lateral plasma membrane domain.

Epithelial cells of the thyroid gland present an uncommon connexin expression pattern, they coexpress connexin32 and connexin43. In the present work, we have analyzed the membrane distribution of these two connexins to determine: (i) whether they co-assemble in the same gap junctions or form separate gap junctions; and (ii) whether their location is somehow related to the thyroid cell polarity. Immunofluorescence analyses of the localization of the two connexins in thyroid tissue sections revealed that connexin32 and connexin43 are located in different regions of the plasma membrane. We further analyzed the location of each of the two connexins with regard to that of the tight junction-associated protein, ZO1. Laser scanning confocal microscope observations of connexin32 or connexin43 and ZO1 double-immunolabelled thyroid cells, gave evidence for a separate localization of gap junctions made of each of these two connexins. Connexin32 gap junctions appeared as fluorescent spots scattered over the lateral membrane domain, while connexin43 gap junctions formed a meshed network superimposable with that of tight junctions in the subapical region of the cells. Western blot analyses of the distribution of connexins in thyroid plasma membrane subfractions obtained by ultracentrifugation on a sucrose gradient led to the identification of membrane sub-populations enriched in either connexin32 gap junctions or connexin43 gap junctions. Connexin32 gap junctions and connexin43 gap junctions were found to differ in their resistance to solubilization by N-lauroylsarcosine. Increasing concentrations of this detergent from 0.12% to 0.42% caused a progressive solubilization of connexin43 while connexin32 remained membrane-bound.(ABSTRACT TRUNCATED AT 250 WORDS)

Incorporation of arachidonic and docosahexaenoic acids into phospholipids of rat brain membranes.

The incorporation of [3H]arachidonic acid (20:4n-6) into rat brain membranes and its mobilization in response to norepinephrine, a relevant neuromediator were studied. The most efficient [3H]20:4n-6 incorporation was in inositol glycerophospholipids (PI) where it reached a plateau after 10 min incubation, while this incorporation was very weak in choline glycerophospholipids (PC). In contrast, the esterification of docosahexaenoic acid, another polyunsaturated fatty acid occurring at high level in brain, was similar in PI and PC, the incorporation in PI being 8-fold lower than that of 20:4n-6. The newly esterified [3H]20:4n-6 was exclusively found in the 1,2-diacyl subclasses of PI and PC. The bulk of incorporation was in the 18:0/20:4n-6 molecular species of 1,2-diacyl-glycerophosphoinositol and in 16:0/20:4n-6 + 18:1/20:4n-6 molecular species of 1,2-diacyl-glycerophosphocholine, which agrees with the usual location of 20:4n-6 in brain phospholipid classes. Upon norepinephrine treatment, [3H]20:4n-6 was not released from PC, but was dose-dependently decreased in PI, the release being significant from 10(-5) M of the agonist. These results suggest that 20:4n-6 exhibits a high specific turnover in brain PI and is mobilized from this class upon relevant neuromediator stimulation. The acellular system used preserved the specificity of enzymes catalyzing the polyunsaturated fatty acid incorporation and release and could be helpful for studying their turn over in brain.

S-adenosyl-l-methionine inhibits phosphoinositide metabolism in the rat brain synaptosomal suspensions.

S-adenosyl-l-methionine (AdoMet) has been reported to affect events linked to noradrenergic neurotransmission. In the present work, we studied the effect of AdoMet on norepinephrine (NE)-stimulated inositol phosphate production in 3H-inositol-labelled crude synaptosomal suspensions of rat brain. AdoMet (50-1000 microM) decreased both the synthesis of labelled polyphosphoinositide (30-50%) and the release of inositol mono- and bisphosphate (40-50%). The AdoMet effect was not dependent on NE concentration (10-1000 microM), suggesting that the inhibition of inositol phosphate release was not the result of a modification of the norepinephrine binding to its receptor sites. S-adenosyl-L-homocysteine (AdoHcy) (1 mM) an inhibitor of methyltransferase activities, partially inhibited (70%) the AdoMet (0.1 mM) effect, indicating that the methylation processes cannot explain all the effects observed. We conclude that, in addition to previously reported effects of AdoMet on NE transport, AdoMet may reduce NE-linked intracellular signalling.

Dynamic analysis of drug action on in vitro reconstituted thyroid follicle by microinjection of tracer molecules and videomicroscopy.

Thyroid cells isolated from the gland by trypsinization are capable in culture of reconstituting histiotypic structures, the thyroid follicles. This morphological differentiation requires the presence of the main thyroid regulator; thyrotropin. We have analyzed some structural and functional aspects of in vitro reconstituted thyroid follicles (RTF) using microinjection of fluorescent probes and videomicroscopy. This experimental approach allowed to visualize biological processes and actions of drugs, signalling factors, etc. in living cells. We describe here some examples of what can be studied with this powerful still-undervalued method. Microinjection of a cell-impermeant fluorescent probe of either high or low molecular mass into the lumen of RTF allowed to check the tightness of this compartment and therefore to analyze the control of tight junctions assembly. A small cell-impermeant probe like Lucifer Yellow microinjected into a cell was used to demonstrate and then to study the regulation of cell to cell communication via gap junctions. The presence of calcium in the lumen of RTF was detected by microinjection of a properly designed probe: Calcium Green which becomes fluorescent in the presence of the ligand. The lumen to cell transport or endocytosis of thyroglobulin, the thyroid prohormone, which is stored into the lumen of the follicles, is currently studied by microinjection of TRITC-labeled thyroglobulin. Coupled to image processing and videorecorder systems, kinetic analysis and quantitative measurements can be performed.

Distribution of tritium labeled 12(S) hydroxy-eicosatetraenoic acid (12-HETE) in the rat.

The in vivo metabolism of 12-(S)-Hydroxy-eicosatetraenoic acid (12-HETE), the end-lipoxygenase product of arachidonic acid in platelets, has been investigated in the rat. Fifty microcuries of 5,6-[3H]-12-HETE (50 Ci/mmol) were injected to anesthetized rats and the radioactivity was followed in plasma. At the end of the experiment, various organs of the animal were removed and the radioactivity attached to them was determined. The label of the plasma plateaued to approximately one third of the initial radioactivity ten minutes after the injection. Among the various organs tested (brain, heart, intestine, kidney, liver, lungs, spleen, testis/uterus) the kidney was far the most active to accumulate 12-HETE and/or its labeled metabolites, and no radioactivity could be detected in urine during the course of the experiment. The analysis of lipid extracts from the various tissues revealed that 12-HETE was not accumulating in its unesterified form but was likely bound to phospholipids. We conclude that, although the label providing from the initial 12-HETE did not completely disappear from plasma, circulating 12-HETE cannot be considered as a circulating marker of cell activation.