Increased production of inflammatory cytokines and activation of microglia in the fetal brain of preeclamptic mice induced by angiotensin II.

Preeclampsia (PE) is a hypertensive obstetric disorder with poor prognosis for both the mother and offspring. Infants born to mothers with PE are known to be at increased risk of developing higher brain dysfunction, such as autism. However, how maternal PE can affect the environment in the fetal brain has not been fully elucidated. Here, we examined the impact of PE on the fetal brain in a mouse model of PE induced by angiotensin II (Ang II), focusing on changes in the inflammatory condition. We confirmed that pregnant mice which were continuously administered Ang II exhibited PE phenotypes, including high blood pressure, proteinuria, and fetal growth restriction. Quantitative RT-PCR analysis demonstrated that the brain of fetuses on embryonic day 17.5 (E17.5) in the Ang II-administered pregnant mice showed increased expression of cytokines, interleukin (IL)- 6, IL-17a, tumor necrosis factor-α, interferon-γ, IL-12, IL-4, and IL-10. Immunohistochemical analysis over a wide area, from the tip of the frontal lobe to the posterior cerebral end, on E17.5 revealed that the microglia in the fetal brain of the Ang II-administered group displayed higher solidity and circularity than those of the control group, indicating that the microglia had transformed to an amoeboid morphology and were activated. Our findings suggest that maternal PE may cause altered inflammatory conditions in the fetal brain, which might be associated with the pathological mechanism connecting maternal PE and brain dysfunction in the offspring.

PKCδ deficiency inhibits fetal development and is associated with heart elastic fiber hyperplasia and lung inflammation in adult PKCδ knockout mice.

Protein kinase C-delta (PKCδ) has a caspase-3 recognition sequence in its structure, suggesting its involvement in apoptosis. In addition, PKCδ was recently reported to function as an anti-cancer factor. The generation of a PKCδ knockout mouse model indicated that PKCδ plays a role in B cell homeostasis. However, the Pkcrd gene, which is regulated through complex transcription, produces multiple proteins via alternative splicing. Since gene mutations can result in the loss of function of molecular species required for each tissue, in the present study, conditional PKCδ knockout mice lacking PKCδI, II, IV, V, VI, and VII were generated to enable tissue-specific deletion of PKCδ using a suitable Cre mouse. We generated PKCδ-null mice that lacked whole-body expression of PKCδ. PKCδ+/- parental mice gave birth to only 3.4% PKCδ-/- offsprings that deviated significantly from the expected Mendelian ratio (χ2(2) = 101.7, P < 0.001). Examination of mice on embryonic day 11.5 (E11.5) showed the proportion of PKCδ-/- mice implanted in the uterus in accordance with Mendelian rules; however, approximately 70% of the fetuses did not survive at E11.5. PKCδ-/- mice that survived until adulthood showed enlarged spleens, with some having cardiac and pulmonary abnormalities. Our findings suggest that the lack of PKCδ may have harmful effects on fetal development, and heart and lung functions after birth. Furthermore, our study provides a reference for future studies on PKCδ deficient mice that would elucidate the effects of the multiple protein variants in mice and decipher the roles of PKCδ in various diseases.

Emotional discrimination during viewing unpleasant pictures: timing in human anterior ventrolateral prefrontal cortex and amygdala.

The ventrolateral prefrontal cortex (VLPFC) and amygdala have critical roles in the generation and regulation of unpleasant emotions, and in this study the dynamic neural basis of unpleasant emotion processing was elucidated by using paired-samples permutation t-tests to identify the timing of emotional discrimination in various brain regions. We recorded the temporal dynamics of blood-oxygen-level-dependent (BOLD) signals in those brain regions during the viewing of unpleasant pictures by using functional magnetic resonance imaging (fMRI) with high temporal resolution, and we compared the time course of the signal within the volume of interest (VOI) across emotional conditions. Results show that emotional discrimination in the right amygdala precedes that in the left amygdala and that emotional discrimination in both those regions precedes that in the right anterior VLPFC. They support the hypotheses that the right amygdala is part of a rapid emotional stimulus detection system and the left amygdala is specialized for sustained stimulus evaluation and that the right anterior VLPFC is implicated in the integration of viscerosensory information with affective signals between the bilateral anterior VLPFCs and the bilateral amygdalae.

Publication criteria for evoked magnetic fields of the human brain: a proposal.

Magnetoencephalography (MEG) is a record of the magnetic fields produced by the electrical activities of the brain using MEG systems. There are three types of sensors for MEG systems: magnetometer and two types of gradiometer. Among them, two types of gradiometer, axial and planar, have been used worldwide. Unfortunately, the waveforms recorded by the two types of gradiometer are often different from each other. This poses a serious problem in comparing and evaluating the data from the two gradiometers. We consider that the MEG study should be published in a way that allows other workers using different types of gradiometer to evaluate and replicate the results of MEG studies. There have been, however, no publication criteria for reports of studies on stimulus-evoked or event-related magnetic fields in human subjects. In this article, we propose publication criteria for evoked or event-related magnetic fields of the human brain: original waveforms of selected channels covering a region of interest, a root mean-squared (RMS) waveform and a contour map at an appropriate time.

Diversity of neural-hemodynamic relationships associated with differences in cortical processing during bilateral somatosensory activation in rats.

The neural-hemodynamic relationships may vary depending on cortical processing patterns. To investigate how cortical hemodynamics reflects neural activity involving different cortical processing patterns, we delivered electrical stimulation pulses to rat hindpaws, unilaterally or bilaterally, and simultaneously measured electrophysiological (local field potential, LFP < 100 Hz; multiunit activity, MUA>300 Hz) and optical intrinsic signals associated with changes in cerebral blood volume (CBV). Unilateral stimulation evoked neural and optical signals in bilateral primary somatosensory cortices. Ipsilateral optical responses indicating an increased CBV exhibited a peak magnitude of ~30% and mediocaudal shifts relative to contralateral responses. Correlation analyses revealed different scale factors between contralateral and ipsilateral responses in LFP-MUA and LFP-CBV relationships. Bilateral stimulation at varying time intervals evoked hemodynamic responses that were strongly suppressed at 40-ms intervals. This suppression quantitatively reflected suppressed LFP responses to contralateral testing stimulation and not linear summation, with slowly fluctuating LFP responses to ipsilateral conditioning stimulation. Consequently, in the overall responses to bilateral stimulation, CBV-related responses were more linearly correlated with MUA than with LFPs. When extracting high-frequency components (>30 Hz) from LFPs, we found similar scale factors between contralateral and ipsilateral responses in LFP-MUA and LFP-CBV relationships, resulting in significant linear relationships among these components, MUA, and cortical hemodynamics in overall responses to bilateral stimulation. The dependence of LFP-MUA-hemodynamic relationships on cortical processing patterns and the LFP temporal/spectral structure is important for interpreting hemodynamic signals in complex functional paradigms driving diverse cortical processing.

Near-infrared spectroscopic study on the effects of chewing on short-term memory.

Using near-infrared spectroscopy, we examined whether chewing gum improves performance in a short-term memory task - immediate recall of random eight-digit numbers - by assessing cerebral hemodynamic response in the prefrontal cortex. We found that the oxyhemoglobin concentration during and after chewing gum was higher than that before chewing; further, the concentration increased during the task, and this increase was reduced with chewing, although non-significantly. Chewing did not improve task performance. Therefore, chewing-induced hemodynamic responses were unrelated to the performance in short-term memory tasks.

Blood oxygenation level-dependent functional magnetic resonance imaging of bilateral but asymmetrical responses to gustatory stimulation in the rat insular cortex.

Evidence has suggested asymmetrical processing of taste in the human insular cortex, but this phenomenon has not been demonstrated in the rodent brain. Functional magnetic resonance imaging (fMRI) is a powerful tool for studying the functional organization of the brain. In this study, we established a blood oxygenation level-dependent (BOLD) fMRI method at 7 T to investigate the responses to gustatory stimulation in the insular cortex of anesthetized rats (220-310 g, n=15). BOLD signals were observed in the insular cortex in response to 0.5 M sucrose solution as the tastant but not observed in response to distilled water as the control. The reproducibility of the BOLD signals in response to the tastant was confirmed between fMRI runs in the same animal and across animals. The signals were mainly located between 2.3 mm and 0.0 mm anterior to the bregma in the insular cortex. Interestingly, the signals were observed in the insular cortex of both hemispheres, but they were asymmetrical: the anterior and posterior regions to the intersection of the middle cerebral artery and the rhinal fissure as the landmark of the gustatory cortex were dominant in the left and right hemispheres of the insular cortex, respectively. These results suggest that activity in both hemispheres of the insular cortex should be considered to analyze taste processing. We think that BOLD fMRI of taste function in rodents will improve our understanding of taste information processing.

Recognition of human emotions from cerebral blood flow changes in the frontal region: a study with event-related near-infrared spectroscopy.

BACKGROUNDS AND PURPOSE: The aim of this study is to develop a near-infrared spectroscopy (NIRS)-based system that recognizes pleasant and unpleasant human emotions based on cerebral blood flow (CBF) in order to understand the minds of patients whose brain function is severely impaired. The forehead region is easily accessible to NIRS measurements, whereas the role of the anterior prefrontal cortex (PFC) in the processing of emotion remains to be elucidated. METHODS: Initially, using event-related NIRS we examined changes in oxygenated hemoglobin (oxy-Hb) as an indicator of regional CBF changes, which reflect brain activity directly related to emotions, but not to cognitive operations in the anterior frontal regions, during viewing affective pictures. The event-related potentials (ERPs), systemic blood pressure, and pulse rate were also measured simultaneously. RESULTS: The event-related analysis of changes in oxy-Hb for a 6 s-picture presentation period showed that very unpleasant emotion was accompanied by an increase in oxy-Hb in the bilateral ventrolateral PFCs, while very pleasant emotion was accompanied by a decrease in oxy-Hb in the left dorsolateral PFC. There were no significant differences in either ERPs or autonomic nervous system activities between the two emotional states. CONCLUSION: These findings suggest the possibility of recognizing patients' emotions from CBF changes.

High-frequency transcutaneous electrical nerve stimulation (TENS) differentially modulates sensorimotor cortices: an MEG study.

OBJECTIVE: Transcutaneous electrical nerve stimulation (TENS) affects excitability of the central motor system as well as the somatosensory system. To determine whether TENS has influence on excitability in the sensorimotor cortices of TENS-treated finger muscle, we investigated magnetoencephalogram associated with voluntary, self-paced finger movement before and after TENS. METHODS: High-frequency TENS was applied on the extensor digitorum muscle for 15 min. Subjects underwent alternate middle finger and thumb extension movements before and after the TENS. We recorded movement-related cortical magnetic field (MRCF) associated with TENS-treated middle finger movement and that from untreated thumb movement. RESULTS: The current source for motor field (MF) was located in the pre-central motor cortex and anteriorly-oriented, and that for motor evoked field one (MEF1) was found in the post-central somatosensory cortex and posteriorly-oriented. The amplitude of MF for TENS-treated middle finger movement decreased but unchanged for untreated thumb movement after TENS. The amplitude of MEF1 decreased for either finger movement after TENS. CONCLUSION: High-frequency TENS to the forearm muscle modulates excitability of the limited area of motor cortex but wider area of primary somatosensory cortex. SIGNIFICANCE: High-frequency TENS to the forearm muscle modulates excitability of the primary somatosensory cortex and motor cortex in a different manner.

Co-activation of the secondary somatosensory and auditory cortices facilitates frequency discrimination of vibrotactile stimuli.

The contribution of the auditory cortex to tactile information processing was studied by measuring somatosensory evoked magnetic fields (SEFs). Three kinds of vibrotactile stimuli with frequencies of 180, 280 and 380 Hz were randomly delivered on the right index finger with a probability of 40, 20 and 40%, respectively. Twenty normal subjects participated in four kinds of tasks: a control condition to ignore these stimuli, a simple task to discriminate the 280-Hz stimulus from the other two stimuli (discrimination task for the vibrotactile stimuli, Ts task), a feedback task modified from the Ts task by adding acoustic feedback of the vibratory frequency at 1300 ms poststimulus (tactile discrimination with auditory clues, TA), and an easy version of the TA task (TA-easy) to discriminate the 280-Hz stimulus (20% target) from the 180- or 380-Hz stimuli (80% nontarget). The Ts and TA tasks required accurate perception of the vibrotactile frequencies to discriminate among the three kinds of stimuli. Under such a task demand, the post hoc auditory feedback in the TA task was expected to induce acoustic imagery for the tactile sensation. The SEFs for the nontarget stimuli were analyzed. A middle-latency component (M150/200) was specifically evoked by the three discrimination tasks. In the Ts and TA-easy tasks, the M150/200 source indicated inferior parietal cortical activities (SII area). In the TA task, 11 subjects showed activity in both the SII area and the superior temporal auditory region and increased accuracy of discrimination compared with the Ts task, in contrast with other subjects who showed activity only in the SII area and small changes in task accuracy between the Ts and TA tasks. Asynchronous auditory feedback for the vibrotactile sensation induced the auditory cortex activity in the SEFs in relation to the progress in tactile discrimination, which suggested an induction of acoustic imagery to complement the tactile information processing.

Intraoperative monitoring of depth-dependent hemoglobin concentration changes during carotid endarterectomy by time-resolved spectroscopy.

By measuring the adult human head during carotid endarterectomy, we investigate the depth sensitivity of two methods for deriving the absorption coefficient changes (Dmu(a)) from time-resolved reflectance data to absorption changes in inhomogeneous media: (1) the curve-fitting method based on the diffusion equation (DE-fit method) and (2) the time-independent calculation based on the modified Lambert-Beer law (MLB method). Remarkable differences in the determined values of Dmu(a) caused by clamping the external carotid artery and subsequently clamping the common carotid artery were observed between the methods. The DE-fit method was more sensitive to mu(a) changes in cerebral tissues, whereas the MLB method was rather sensitive to mu(a) changes in the extracerebral tissues. Our results indicated that the DE-fit was useful for monitoring the cerebral blood circulation and oxygenation during neurosurgical operations. In addition, the combined evaluation of mu(a) changes with the DE-fit and MLB methods will provide us with more available information about the hemodynamic changes in the depth direction.

Attention induces reciprocal activity in the human somatosensory cortex enhancing relevant- and suppressing irrelevant inputs from fingers.

OBJECTIVE: We studied whether attention regulates information processing in the human primary somatosensory cortex (SI) by selective enhancement of relevant- and suppression of irrelevant information. METHODS: Under successive and simultaneous electric stimuli to both the right index and middle fingers, tactile stimuli were randomly (20%) presented on one of the two fingers in separate two runs exchanging the finger. Subjects were requested to discriminate the tactile stimuli in an attention task to induce attention to one finger and to ignore the stimuli in a control task to avoid such an attention focus. Somatosensory evoked magnetic fields were measured only for the two-finger electric stimulation and an early component (M50) was analyzed. RESULTS: In spite of the two-finger simultaneous stimulation, attention to either the index or middle finger lowered or heightened the M50-sourse location, respectively. The attention task did not increase the M50 amplitude. CONCLUSIONS: Attention to a finger enhanced selectively the representation of the finger in the SI cortex. However, this SI activity did not increase the M50 amplitude, suggesting that the attention suppressed another finger region receiving the unattended inputs. SIGNIFICANCE: Attention regulates the SI activity by selectively enhancing the task-relevant information and by filtering out other noise inputs.

Reevaluation of near-infrared light propagation in the adult human head: implications for functional near-infrared spectroscopy.

Using both experimental and theoretical methods, we examine the contribution of different parts of the head to near-IR (NIR) signal. Time-resolved spectroscopy is employed to measure the mean optical path length (PL), and the absorption (mu(a)) and reduced scattering (mu(s)') coefficients in multiple positions of the human head. Monte Carlo simulations are performed on four-layered head models based on an individual magnetic resonance imaging (MRI) scan to determine mu(a) and mu(s)' in each layer of the head by solving inverse problems, and to estimate the partial path length in the brain (p-PL) and the spatial sensitivity to regions in the brain at the source-detector separation of 30 mm. The PL is closely related to the thickness of the scalp, but not to that of other layers of the head. The p-PL is negatively related to the PL and its contribution ratio to the PL is 5 to 22% when the differential path length factor is 6. Most of the signal attributed to the brain comes from the upper 1 to 2 mm of the cortical surface. These results indicate that the NIR signal is very sensitive to hemodynamic changes associated with functional brain activation in the case that changes in the extracerebral tissue are ignorable.

Functional connectivity between forearm and digits representations in human somatosensory area 3b.

OBJECTIVE: We compared the effects of tactile interference to the forearm on magnetic responses evoked by electric stimulation of the little finger (D5) and the thumb (D1). METHODS: Electric stimulation was delivered to D5 or D1 individually. In each stimulus session, magnetic recordings were conducted with or without concurrent tactile interference to the radial side of the anterior forearm. RESULTS: With forearm interference, the amplitude of the primary response (N20m) following D5 stimulation was reduced to 90.7% of the control value without interference, while that following D1 stimulation was not affected (100.7%). CONCLUSIONS: In human somatosensory area 3b, the representation of the forearm is immediately adjacent to that of the D5, and distant from that of the D1. Thus, the result suggests that the tactile interference effect on N20m depends on the cortical distance between electrically and mechanically activated 3b areas. SIGNIFICANCE: Intrinsic synaptic connections between the 3b hand representation and its surroundings have been hypothesized as a neural basis for plastic changes of the human brain, such as a phantom hand phenomenon. The present finding implies that these connections may play some physiological roles even in normal adult humans.

Is there training-dependent reorganization of digit representations in area 3b of string players?

OBJECTIVE: The digit representations in area 3b were studied to examine whether there is training-dependent reorganization in string players. METHODS: Somatosensory evoked magnetic fields were recorded following electrical stimulation of digits 1 (D1), 2 (D2) and 5 (D5) of both hands in 8 string players and of the left hand in 12 control subjects. The N20m and P30m responses, and high-frequency oscillations (HFOs) were separated by 3-300 Hz and 300-900 Hz bandpass filtering. RESULTS: The dipole locations on the coronal plane and strengths of D1, D2 and D5, and D1-D5 cortical distance estimated at the peak of N20m or P30m did not differ between left and right hand in string players or between left hand in string players and controls. On the other hand, the dipole locations of D2 estimated from N20m and P30m and of D1 from N20m were significantly anterior, the D2-D5 distance from P30m longer, and the number of HFO peaks larger for D5 in string players than controls. CONCLUSIONS/SIGNIFICANCE: With strong mutual competition among the fingering digits, the scale of reorganization should be much smaller as compared with the competition-free denervation-induced reorganizations. Taken together, the training-dependent reorganization of somatosensory cortex in string players is manifest not only in the enlarged cortical representation but also in the enhanced HFOs presumably representing activity of the fast-spiking interneurons.

Spatiotemporal characteristics of hemodynamic changes in the human lateral prefrontal cortex during working memory tasks.

The prefrontal cortex (PFC) is widely believed to subserve mental manipulation and monitoring processes ascribed to the central executive (CE) of working memory (WM). We attempted to examine and localize the CE by functional imaging of the frontal cortex during tasks designed to require the CE. Using near-infrared spectroscopy, we studied the spatiotemporal dynamics of oxygenated hemoglobin (oxy-Hb), an indicator of changes in regional cerebral blood flow, in both sides of lateral PFC during WM intensive tasks. In most participants, increases in oxy-Hb were localized within one subdivison during performance of the n-back task, whereas oxy-Hb increased more diffusely during the random number generation (RNG) task. Activation of the ventrolateral PFC (VLPFC) was prominent in the n-back task; both sustained and transient dynamics were observed. Transient dynamics means that oxy-Hb first increases but then decreases to less than 50% of the peak value or below the baseline level before the end of the task. For the RNG task sustained activity was also observed in the dorsolateral PFC (DLPFC), especially in the right hemisphere. However, details of patterns of activation varied across participants: subdivisions commonly activated during performance of the two tasks were the bilateral VLPFCs, either side of the VLPFC, and either side of the DLPFC in 4, 2, and 4 of the 12 participants, respectively. The remaining 2 of the 12 participants had no regions commonly activated by these tasks. These results suggest that although the PFC is implicated in the CE, there is no stereotyped anatomical PFC substrate for the CE.

Effect of stimulus frequency on human cerebral hemodynamic responses to electric median nerve stimulation: a near-infrared spectroscopic study.

We examined the effect of stimulus frequency on optically recorded hemodynamic responses to electric median nerve stimulation. Electric stimuli were delivered to the right median nerve with an intensity of 90% of motor threshold. Four different stimulus frequencies (2, 5, 10, and 20 Hz) were administered in each subject. By means of a multi-channel near-infrared spectroscopic instrument, changes in concentration of oxygenated hemoglobin were continuously measured over the left scalp. After 20 Hz stimulation, we found two spatially and temporally distinct hemodynamic responses. One lasted beyond 60 s, and the center of this response was located over the secondary somatosensory area. The other had a transient duration starting immediately after the stimulus onset and was located in the primary somatosensory hand area. Both responses were linearly augmented as a function of the stimulus frequency. Since temporal activation patterns are different in two somatosensory areas, real-time optical monitoring is necessary in evaluation of hemodynamic responses to electric nerve stimulation.

Muscle afferent inputs from the hand activate human cerebellum sequentially through parallel and climbing fiber systems.

OBJECTIVE: Spatio-temporal response characteristics of the human cerebellum to median nerve stimulation (MNS) were studied with the use of a whole-head magnetoencephalographic (MEG) system covering the cerebellum and upper cervical spine. METHODS: Neuromagnetic responses from the cerebellum were recorded following electric stimulation of the right median nerve in 12 subjects. In 6 out of 12 subjects, the responses to the left median nerve and to the right index or middle finger stimulation were also recorded. RESULTS: The medial part of the cerebellum (spinocerebellum) was activated by MNS. In contrast, there were no responses from the cerebellum to the finger stimulation, suggesting that muscle afferent inputs are the source of cerebellar activation for MNS. The cerebellar responses consisted of 3 or 4 components of alternating polarity within 90 ms post-stimulus: the current direction for the first component was from the depth to the surface of the anterior lobe. CONCLUSIONS: From the timing and current direction, we speculate that the 4 components reflect, respectively, (1) excitatory postsynaptic potentials (EPSPs) of granule cells, (2) Purkinje cell EPSPs at the distal dendrites driven by parallel fibers, (3) Purkinje cell EPSPs at the soma and the proximal dendrites mediated by climbing fibers and (4) second Purkinje cell EPSPs at the distal dendrites driven by parallel fibers. SIGNIFICANCE: We first visualized serial activation of the human spinocerebellum following MNS noninvasively with MEG.

Tactile interference to the face affects magnetic responses elicited by electric thumb stimulation.

OBJECTIVE: We examined the effect of tactile interference to the face on somatosensory evoked magnetic fields (SEFs) following electric thumb stimulation. METHODS: SEFs were elicited by electric stimulation of the right thumb in a control and two interference conditions. In the interference conditions, continuous tactile stimuli were delivered to the skin surface over the right upper face or the right thumb. RESULTS: The face interference significantly attenuated N20m and enhanced P30m. The amplitudes of N20m in the face and thumb interference conditions were 90.3 and 70.3% of the value in the control condition, respectively, while those of P30m were 120.2 and 74.4%. CONCLUSIONS: In human somatosensory area 3b, the representation of the thumb is immediately adjacent to that of the face although the thumb and face are physically distant. We suggest, therefore, that the effect of tactile interference on N20m depends on a cortical distance between electrically and mechanically activated 3b areas, rather than a physical distance between the body parts to which these two stimuli were administered. Although it is unclear why the face interference specifically enhanced the P30m, it is suggested that the generating mechanism of the interference effect on P30m may be different from that on N20m. SIGNIFICANCE: The tactile interference effect on N20m does not depend on the physical distance between electrically and mechanically activated skin areas, but on the distance of the 3b cortex receiving these two inputs.

Selective attention regulates spatial and intensity information processing in the human primary somatosensory cortex.

Attention-related cognitive processes in the primary somatosensory cortex (SI) were studied by measuring somatosensory evoked magnetic fields (SEFs). Twenty-one normal adult human subjects participated in this study for investigating effects of attention and stimulus intensity on cortical finger representation in the SI cortex. Electric stimuli at low and high intensity were delivered to the index or middle finger in finger discrimination and non-discrimination task. For the low intensity stimulation at 1.25 times sensory threshold, an early component (M50) showed clear segregation of the sources for the two fingers and an increase of the amplitude specific to the finger discrimination task. Such an attentional effect on the SI cortex was masked by the high intensity stimulation (2.5 times sensory threshold); the M50 source separation by the fingers was induced irrespective of the discrimination or non-discrimination task. The results suggest that a conscious regulation of stimulus intensity coding in the SI cortex underlies the attention-dependent enhancement of spatial finger information processing.