Susumu Urakawa

Rearing in enriched environment increases parvalbumin-positive small neurons in the amygdala and decreases anxiety-like behavior of male rats.

BackgroundEarly life experiences including physical exercise, sensory stimulation, and social interaction can modulate development of the inhibitory neuronal network and modify various behaviors. In particular, alteration of parvalbumin-expressing neurons, a gamma-aminobutyric acid (GABA)ergic neuronal subpopulation, has been suggested to be associated with psychiatric disorders. Here we investigated whether rearing in enriched environment could modify the expression of parvalbumin-positive neurons in the basolateral amygdala and anxiety-like behavior.ResultsThree-week-old male rats were divided into two groups: those reared in an enriched environment (EE rats) and those reared in standard cages (SE rats). After 5 weeks of rearing, the EE rats showed decreased anxiety-like behavior in an open field than the SE rats. Under another anxiogenic situation, in a beam walking test, the EE rats more quickly traversed an elevated narrow beam. Anxiety-like behavior in the open field was significantly and negatively correlated with walking time in the beam-walking test. Immunohistochemical tests revealed that the number of parvalbumin-positive neurons significantly increased in the basolateral amygdala of the EE rats than that of the SE rats, while the number of calbindin-D28k-positive neurons did not change. These parvalbumin-positive neurons had small, rounded soma and co-expressed the glutamate decarboxylase (GAD67). Furthermore, the number of parvalbumin-positive small cells in the basolateral amygdala tended to positively correlate with emergence in the center arena of the open field and negatively correlated with walking time in the beam walking test.ConclusionRearing in the enriched environment augmented the number of parvalbumin-containing specific inhibitory neuron in the basolateral amygdala, but not that of calbindin-containing neuronal phenotype. Furthermore, the number of parvalbumin-positive small neurons in the basolateral amygdala was negatively correlated with walking time in the beam walking test and tended to be positively correlated with activity in the center arena in the open field test. The results suggest that rearing in the enriched environment augmented parvalbumin-positive specific neurons in the basolateral amygdala, which induced behavioral plasticity that was reflected by a decrease in anxiety-like behavior in anxiogenic situations.

Cognitive and socio-emotional deficits in platelet-derived growth factor receptor-β gene knockout mice.

Platelet-derived growth factor (PDGF) is a potent mitogen. Extensive in vivo studies of PDGF and its receptor (PDGFR) genes have reported that PDGF plays an important role in embryogenesis and development of the central nervous system (CNS). Furthermore, PDGF and the β subunit of the PDGF receptor (PDGFR-β) have been reported to be associated with schizophrenia and autism. However, no study has reported on the effects of PDGF deletion on mice behavior. Here we generated novel mutant mice (PDGFR-β KO) in which PDGFR-β was conditionally deleted in CNS neurons using the Cre/loxP system. Mice without the Cre transgene but with floxed PDGFR-β were used as controls. Both groups of mice reached adulthood without any apparent anatomical defects. These mice were further examined by conducting several behavioral tests for spatial memory, social interaction, conditioning, prepulse inhibition, and forced swimming. The test results indicated that the PDGFR-β KO mice show deficits in all of these areas. Furthermore, an immunohistochemical study of the PDGFR-β KO mice brain indicated that the number of parvalbumin (calcium-binding protein)-positive (i.e., putatively γ-aminobutyric acid-ergic) neurons was low in the amygdala, hippocampus, and medial prefrontal cortex. Neurophysiological studies indicated that sensory-evoked gamma oscillation was low in the PDGFR-β KO mice, consistent with the observed reduction in the number of parvalbumin-positive neurons. These results suggest that PDGFR-β plays an important role in cognitive and socioemotional functions, and that deficits in this receptor may partly underlie the cognitive and socioemotional deficits observed in schizophrenic and autistic patients.

Compression on trigger points in the leg muscle increases parasympathetic nervous activity based on heart rate variability

Massotherapy, the therapeutic use of massage, is used to treat various chronic pain syndromes. One type of massotherapy, pressure stimulus applied over trigger points (TPs), is reported to have excellent therapeutic effects. Its effect is possibly mediated through changes in the autonomic nervous system although little research has been conducted to assess autonomic activity during TP compression. We have investigated how compression applied over TPs affects the autonomic nervous system. Six healthy young adult females whose daily working routine was carried out predominantly in a standing position were enrolled in the study cohort. After a days work, the subjects were asked to rest supine, and electrocardiograms (ECGs), instantaneous lung volume (ILV) and systolic and diastolic blood pressures (SBP, DBP) were measured before and after pressure application over the TPs in those lower limb muscles where the subjects felt muscle fatigue or discomfort. The subjects were also asked to coordinate breathing with the beeping sounds. The therapeutic effects of TP compression were assessed by a subjective fatigue scale. Parasympathetic nervous activity was also assessed by spectral analysis of heart rate (HR) variability. The transfer function from ILV to HR was evaluated using linear analysis. The results indicated that TP compression (1) decreased HR, SBP and DBP, (2) increased parasympathetic activity, (3) increased the gain from ILV to HR, and (4) improved the fatigue scores. These findings suggest that an increase in parasympathetic nervous activity after the TP compression induced a reduction of fatigue. The therapeutic mechanisms of TP compression to enhance parasympathetic nervous system are discussed.

Effects of acupuncture on the brain hemodynamics

Acupuncture therapy has been applied to various psychiatric diseases and chronic pain since acupuncture stimulation might affect brain activity. From this point of view, we investigated the effects of acupuncture on autonomic nervous system and brain hemodynamics in human subjects using ECGs, EEGs and near-infrared spectroscopy (NIRS). Our previous studies reported that changes in parasympathetic nervous activity were correlated with number of de-qi sensations during acupuncture manipulation. Furthermore, these autonomic changes were correlated with EEG spectral changes. These results are consistent with the suggestion that autonomic changes induced by needle manipulation inducing specific de-qi sensations might be mediated through the central nervous system, especially through the forebrain as shown in EEG changes, and are beneficial to relieve chronic pain by inhibiting sympathetic nervous activity. The NIRS results indicated that acupuncture stimulation with de-qi sensation significantly decreased activity in the supplementary motor complex (SMC) and dorsomedial prefrontal cortex (DMPFC). Based on these results, we review that hyperactivity in the SMC is associated with dystonia and chronic pain, and that in the DMPFC is associated with various psychiatric diseases with socio-emotional disturbances such as schizophrenia, attention deficit hyperactive disorder, etc. These findings along with the previous studies suggest that acupuncture with de-qi sensation might be effective to treat the various diseases in which hyperactivity in the SMA and DMPFC is suspected of playing a role.

Significant correlation between autonomic nervous activity and cerebral hemodynamics during thermotherapy on the neck

Although local thermotherapy reduces mental stress and neck stiffness, its physiological mechanisms are still not fully understood. We speculated that local thermotherapy exerts its effect, in addition to its direct peripheral effects, through the central nervous system that is involved in controlling stress responses. In the present study, we investigated the effects of a heat- and steam-generating (HSG) sheet on cerebral hemodynamics and autonomic nervous activity using near-infrared spectroscopy (NIRS) and the electrocardiograms (ECGs). Thirteen healthy young female subjects participated in this study. HSG or simple (control) sheets were repeatedly applied to the neck for 120 s with 180 s intervals of rest between applications. During the experiment, brain hemodynamic responses (changes in Oxy-Hb, Deoxy-Hb, and Total-Hb) and autonomic nervous activity based on heart rate variability (HRV) were monitored. Subjective perception of neck stiffness and fatigue was significantly improved after application of the HSG sheet. NIRS findings indicated that the application of HSG sheets decreased Oxy-Hb concentration in the anterior-dorsal region of the medial prefrontal cortex (adMPFC), while increasing parasympathetic nervous activity and decreasing sympathetic nervous activity. Furthermore, changes in Oxy-Hb in the adMPFC were significantly and negatively correlated with those in parasympathetic nervous activity during application of the HSG sheet. These findings suggest that application of the HSG sheet to the neck region induced mental relaxation and ameliorated neck stiffness by modifying activity of the adMPFC.

A 3D-Video-Based Computerized Analysis of Social and Sexual Interactions in Rats

A large number of studies have analyzed social and sexual interactions between rodents in relation to neural activity. Computerized video analysis has been successfully used to detect numerous behaviors quickly and objectively; however, to date only 2D video recording has been used, which cannot determine the 3D locations of animals and encounters difficulties in tracking animals when they are overlapping, e.g., when mounting. To overcome these limitations, we developed a novel 3D video analysis system for examining social and sexual interactions in rats. A 3D image was reconstructed by integrating images captured by multiple depth cameras at different viewpoints. The 3D positions of body parts of the rats were then estimated by fitting skeleton models of the rats to the 3D images using a physics-based fitting algorithm, and various behaviors were recognized based on the spatio-temporal patterns of the 3D movements of the body parts. Comparisons between the data collected by the 3D system and those by visual inspection indicated that this system could precisely estimate the 3D positions of body parts for 2 rats during social and sexual interactions with few manual interventions, and could compute the traces of the 2 animals even during mounting. We then analyzed the effects of AM-251 (a cannabinoid CB1 receptor antagonist) on male rat sexual behavior, and found that AM-251 decreased movements and trunk height before sexual behavior, but increased the duration of head-head contact during sexual behavior. These results demonstrate that the use of this 3D system in behavioral studies could open the door to new approaches for investigating the neuroscience of social and sexual behavior.

Neuronal Responses in the Nucleus Accumbens Shell during Sexual Behavior in Male Rats

Previous behavioral studies have indicated that the nucleus accumbens (NAc) shell of a male rat is involved in its sexual behavior; however, no previous studies have investigated neuronal activities in the male rat NAc shell during sexual behavior. To investigate this issue, we recorded single unit activities in the NAc shell of male rats during sexual behavior. Of 123 NAc shell neurons studied, 53, 47, and 40 neurons exhibited significantly changed firing rates at various times during intromission, genital auto-grooming, and sniffing of females, respectively. The two types of NAc shell neurons [putative fast spiking interneurons (pFSIs) and medium spiny neurons (pMSNs)] responded differently during sexual behavior. First, more pFSIs than pMSNs exhibited inhibitory responses to thrusting with intromission and genital grooming, while pFSIs and pMSNs responded similarly to sniffing of females. Second, both pFSIs and pMSNs responded differently to thrusting with and without intromission. Furthermore, NAc shell neuronal activity was significantly different across the different phases of sexual behavior, and the number of NAc shell neurons with delta oscillation, which is related to behavioral inhibition, and high gamma oscillation, which is related to reward perception, increased after ejaculation. Together, our results suggest that the NAc shell is deeply involved in sexual behavior, and changes in NAc shell neuronal activity are related to performance of sexual behavior, encoding cues or contexts related to sexual behavior, reward-related processing, and the inhibition of sexual behavior after ejaculation.

Olfactory preference in the male rat depends on multiple chemosensory inputs converging on the preoptic area.

Both volatile and nonvolatile molecules are involved in chemosensory communication in rodents. Volatile odors from physically inaccessible estrous females induced increased numbers of c-Fos-positive cells in the preoptic area (POA) and in the cortical nucleus of the amygdala (CoA) of male rats. The numbers of c-Fos-positive cells in the medial nucleus of the amygdala (MeA) increased in response to the nonvolatile odors of bedding soiled with the excreta of estrous females. In an alternate choice paradigm, male rats carrying ibotenic acid lesions in either the MeA or the CoA--or a combination of both--distinguished the odors of estrous females from those of males, although the time spent sniffing the stimuli was diminished. Males with POA lesions showed complete loss of this capability. Males carrying either of the lesions did not detect differences between estrous and anestrous females or between intact and orchidectomized males. Lesions in the POA or MeA severely impaired male sexual behavior, whereas a CoA lesion had no effects. Thus, c-Fos-positive cells in the CoA might be involved in chemosensory transmission relevant to certain social contexts, but not in the execution of male sexual behavior. The POA is indispensable for both olfactory preferences and sexual behavior. The residual olfactory preference in males with MeA or CoA lesions or the combination of both could reflect an additional route for chemosensory transmission from the main olfactory bulb to the POA.

Cerebral functional imaging using near-infrared spectroscopy during repeated performances of motor rehabilitation tasks tested on healthy subjects

To investigate the relationship between the frontal and sensorimotor cortices and motor learning, hemodynamic responses were recorded from the frontal and sensorimotor cortices using functional near infrared spectroscopy (NIRS) while healthy subjects performed motor learning tasks used in rehabilitation medicine. Whole-head NIRS recordings indicated that response latencies in the anterior dorsomedial prefrontal cortex (aDMPFC) were shorter than in other frontal and parietal areas. Furthermore, the increment rate of the hemodynamic responses in the aDMPFC across the eight repeated trials significantly correlated with those in the other areas, as well as with the improvement rate of task performance across the 8 repeated trials. In the second experiment, to dissociate scalp- and brain-derived hemodynamic responses, hemodynamic responses were recorded from the head over the aDMPFC using a multi-distance probe arrangement. Six probes (a single source probe and 5 detectors) were linearly placed 6 mm apart from each of the neighboring probes. Using independent component analyses of hemodynamic signals from the 5 source-detector pairs, we dissociated scalp- and brain-derived components of the hemodynamic responses. Hemodynamic responses corrected for scalp-derived responses over the aDMPFC significantly increased across the 8 trials and correlated with task performance. In the third experiment, subjects were required to perform the same task with and without transcranial direct current stimulation (tDCS) of the aDMPFC before the task. The tDCS significantly improved task performance. These results indicate that the aDMPFC is crucial for improved performance in repetitive motor learning.

Global synchronization in the theta band during mental imagery of navigation in humans.

Visual mental imagery is critical for successfully navigating the environment, which in turn activates many cortical regions simultaneously. Theta oscillation is implicated in navigation and brain synchronization. In this study, EEG coherence was analyzed during 3 tasks: subjects (1) mentally simulated jogging along the walls of a gym and pressed a button when they imagined arriving at a corner (jogging imagery task), (2) thought of and memorized one digit after pressing a button 5 times and recalled the digits sequentially after pressing the button again (digit imagery task), and (3) pressed a button (button pressing task). The results indicated that theta-wave (4-8 Hz) power was significantly higher in the frontal and parietal regions during the digit and jogging imagery tasks. Coherence at the theta band showed almost no differences between the button pressing and digit imagery tasks. Coherence between the distant regions, especially between the frontal and parieto-occipital regions and between interhemispheric regions, was significantly higher during the jogging imagery task. Increase in theta power during the jogging imagery task reflects working memory load to manipulate internal information. Theta oscillation appears to play an important role in large-scale synchronization to form the functional neuronal networks required for mental navigation.