Ichiro Takashima

Voltage-sensitive dye versus intrinsic signal optical imaging : comparison of optically determined functional maps from rat barrel cortex

Using intrinsic and voltage-sensitive dye optical imaging methods, somatosensory-evoked neural activity and the consequent metabolic activity were visualized in the barrel cortex at high temporal and spatial resolution. We compared maps of neural and metabolic activity from the perspective of spatial distribution in the cortex. There was good agreement between the two functional maps, if the extent of metabolic activity before a prominent increase in cerebral blood volume (CBV) was assessed. This result indicates that oxygen consumption occurs before CBV changes, in approximately the same cortical area as that in which the preceding neural activity was evoked. This also suggests that the intrinsic signal reflects subthreshold synaptic activity, as well as spiking activity, which is similar to the dye-related signals.

High-speed CCD imaging system for monitoring neural activity in vivo and in vitro, using a voltage-sensitive dye

We have designed and constructed a high-speed CCD imaging system for optically detecting neural activity from preparations stained externally with a voltage-sensitive dye, and have used this system to image evoked and epileptiform neural activity in the rat somatosensory cortex. The imaging system uses a commercially available 1/3-in. CCD chip, and it can continuously capture images for more than 8 s, at 1000 frames/s, with a spatial resolution of 128 x 62 pixels. The spatial/temporal resolution of the CCD sensor is variable by changing the geometry of on-chip binning pixels, which can be controlled by a PC/AT computer. Dye bleaching correction was not necessary for long-term imaging of epileptiform neural events, since the sensitivity of the CCD sensor was increased by combining the signal from adjacent pixels.

Optical recording of cortical activity after in vitro perfusion of cerebral arteries with a voltage-sensitive dye.

Cortical neuronal architecture and connectivity can be analyzed with high-resolution optical imaging after staining the in vitro isolated guinea pig brain preparation by circulating the voltage-sensitive dye RH795 via the arterial system. To establish this new technique, electrical field potentials evoked in the piriform and entorhinal cortices by lateral olfactory tract stimulation were correlated to the optical signal. The depth analysis of the optical response was performed by evaluating the contribution of the mono- and poly-synaptic components of the signal generated in different layers after applying a pair-pulse stimulation protocol. The tangential propagation of neuronal activity in olfactory cortices was evaluated by gathering several 4.2 x 4.2 mm images recorded from adjacent cortical areas. The real-time optical imaging technique applied to the isolated guinea pig brain can be successfully utilized to study the integrative properties of cortical neurons ensembles.

Olfactory information converges in the amygdaloid cortex via the piriform and entorhinal cortices: observations in the guinea pig isolated whole-brain preparation

The amygdaloid cortex (AC) has reciprocal connections with the entorhinal cortex (EC) and also receives projections from the olfactory bulb and the piriform cortex (PC). To assess the possibility that the AC and EC represent functionally coupled structures in the olfactory stream of information, we investigated the propagation pattern of neural activity in olfactory cortices − PC, AC and EC − using optical recordings with voltage‐sensitive dyes in the guinea pig in vitro isolated whole‐brain preparation. We observed two distinct pathways that convey neural activation evoked by olfactory nerve stimulation: a medial pathway from the PC to the AC, and a lateral pathway from the PC to the lateral EC along the rhinal sulcus. Besides being activated directly via the medial pathway, the AC was activated a second time via activity that propagated from the lateral EC. Lesion experiments revealed that the lateral pathway close to the rhinal sulcus is crucial for neural activation of the EC. Consistent with this activation pattern, we observed two separate, sharp downward deflections in field potential recordings, and we recorded synaptic potentials with multiple peaks from single neurons in the AC. Our findings suggest that the AC and EC are functionally coupled during olfactory information processing, and that this functional linkage may allow the AC to integrate olfactory sensation with information retained or processed in the EC.

Architecture of odor information processing in the olfactory system

Since the discovery of the superfamily of approximately 1000 odorant receptor genes in rodents, the structural simplicity as well as the complexity of the olfactory system have been revealed. The simple aspects include the one neuron-one receptor rule and the exclusive convergence of projections from receptor neurons expressing the same receptors to one or two glomeruli in the olfactory bulb. Odor decoding in the olfactory cortex or higher cortical areas is likely to be a complicated process that depends on the sequence of signal activation and the relative signal intensities of receptors overlapping for similar but different odors. The aim of the present study was to investigate odor information processing both in receptors and in the olfactory cortex. At the receptor level, the similarity and difference in receptor codes between a pair of chiral odorants were examined using the tissue-printing method for sampling all the epithelial zones. In order to dissect odor-driven signal processing in the olfactory cortex by reducing cross-talk with the non-olfactory activities, such as cyclic respiration or other sensory inputs, an in vitro preparation of isolated whole brain with an attached nose was developed, and the methodologies and resulting hypothesis of receptor-sensitivity-dependent hierarchical odor information coding were reviewed.

Voltage-sensitive dye imaging of intervibrissal fur-evoked activity in the rat somatosensory cortex

The intervibrissal fur-evoked activity in the rat somatosensory cortex was investigated using high-resolution optical imaging with a voltage-sensitive dye. The optical imaging revealed that the intervibrissal fur representation forms a U-shaped band around the borders of the posteromedial barrel subfield (PMBSF), and that this representation is characterized by a rostral-to-caudal somatotopic organization. When GABA(A)-mediated inhibition was partially suppressed by treatment with bicuculline, stimulation of the intervibrissal fur elicited spreading of an excitation wave in an area outside the PMBSF. The spreading wave propagated in both directions along the aforementioned U-shaped band of cortex, but barely invaded the center of the PMBSF. These imaging results suggest a distinct subdivision of cortex adjacent to, but outside, the PMBSF in the rat somatosensory cortex; this region receives input from intervibrissal fur, and seems to process its sensory information through well-developed local horizontal connections.

Optical imaging of rat prefrontal neuronal activity evoked by stimulation of the ventral tegmental area.

Using a voltage-sensitive dye, the spatiotemporal dynamics of prefrontal neuronal activity evoked by electrical stimulation of the ventral tegmental area were visualized through optical imaging in anaesthetized rats. Even single-pulse stimulation of the ventral tegmental area elicited a widespread wave of depolarization followed by hyperpolarization in the dorsomedial shoulder region of the prefrontal cortex. We also examined the contribution of dopaminergic transmission to the optical signals by comparing normal and 6-hydroxydopamine-lesioned rats. The 6-hydroxydopamine lesions of ventral tegmental area resulted in a complete absence of depolarization in the prefrontal cortex, although hyperpolarization was preserved. These results indicate that dopaminergic neurons are needed to generate excitatory responses in the prefrontal cortex.

A novel method for quantifying similarities between oscillatory neural responses in wavelet time-frequency power profiles.

Quantifying similarities and differences between neural response patterns is an important step in understanding neural coding in sensory systems. It is difficult, however, to compare the degree of similarity among transient oscillatory responses. We developed a novel method of wavelet correlation analysis for quantifying similarity between transient oscillatory responses, and tested the method with olfactory cortical responses. In the anterior piriform cortex (aPC), the largest area of the primary olfactory cortex, odors induce inhibitory activities followed by transient oscillatory local field potentials (osci-LFPs). Qualitatively, the resulting time courses of osci-LFPs for identical odors were modestly different. We then compared several methods for quantifying the similarity between osci-LFPs for identical or different odors. Using fast Fourier transform band-pass filters, a conventional method demonstrated high correlations of the 0-2Hz components for both identical and different odors. None of the conventional methods tested demonstrated a clear correlation between osci-LFPs. However, wavelet correlation analysis resolved a stimulus dependency of 2-45Hz osci-LFPs in the aPC output layer, and produced experience-dependent high correlations in the input layer between some of the identical or different odors. These results suggest that redundancy in the neural representation of sensory information may change in the aPC. This wavelet correlation analysis may be useful for quantifying the similarities of transient oscillatory neural responses.

Impairment of the discrimination of the direction of single-whisker stimulation induced by the lemniscal pathway lesion

In the rodent somatosensory system, stimulus information received by the whiskers is relayed to the barrel cortex via two parallel pathways, the lemniscal pathway and the paralemniscal pathway. The lemniscal pathway includes the principal trigeminal nucleus (Pr5) and the ventral posteromedial thalamic nucleus (VPm). The paralemniscal pathway includes the spinal trigeminal subnucleus interpolaris (Sp5i) and the medial division of posterior thalamic nucleus (POm). The purpose of this study was to investigate the roles of those pathways in perceptions of the direction of the single-whisker stimulation in the rat. Rats were trained to perform a go/no-go task that required the discrimination of forward or backward stimulation applied to their single whisker. When a selective lesion was made in VPm or Pr5, error rate for the task performance increased significantly. In contrast, when a selective lesion was made in POm or Sp5i, we found no significant change in performance. These results suggest that the lemniscal pathway plays more important roles in a discrimination of stimulus direction applied to the single whisker.

Blood-brain barrier derangement after electrical brain stimulation

Noninvasive brain stimulation methods, including repetitive transcranial magnetic stimulation and transcranial direct current stimulation (tDCS), have received considerable attention in recent years for use in the study and treatment of neurological conditions. Of these methods, tDCS is considered particularly promising due to its ease of use and ability to confer polarity-dependent effects on brain excitability, making it an excellent option for clinical treatment of neurological and psychiatric diseases. While generally regarded as safe when following standard protocols, the effects of tDCS on cerebral blood vessels and blood-brain barrier (BBB) functions remain poorly understood. Here, we provide an overview of tDCS in the context of BBB function, summarize the current literature, and discuss implications for future research. To date, no alterations or damage to the BBB have been reported after weak tDCS stimulations in human subjects; however, some animal studies have reported alterations to BBB function following increased tDCS intensity, with inconsistencies in the effective tDCS polarity used to produce these BBB disruptions between studies. Further research will be necessary to evaluate the effects of tDCS on the BBB under various conditions. Finally, we discuss the potential of tDCS for enhancing drug delivery to the central nervous system, which may become possible as we refine our understanding of the effects of tDCS on BBB permeability.