Teppei Noda

Response properties and morphological identification of neurons in the cat motor cortex

Intracellular recordings and morphological identification of neurons using intracellular HRP staining were performed in the cat motor cortex. By thalamic ventrolateral (VL) or cerebellar nucleus stimulation, pyramidal cells in layer III, fast pyramidal tract neurons (PTNs) and stellate cells in layers II and III were activated with short latency and fast rising EPSPs, while pyramidal cells in layer II and slow PTNs showed longer latency and slow rising EPSPs. This difference may be related to activation through the deep and superficial thalamocortical projections. Although pyramidal cells in layer VI did not respond orthodromically to VL or cerebellar stimulation, some of them proved to receive the recurrent action of PTNs because of the response to stimulation of the cerebral peduncle (CP). One aspinous stellate cell in layer III was activated by CP as well as VL stimulation. This cell was supposed to be an inhibitory interneuron responsible for both recurrent and VL-evoked inhibition.

Restriction of Wnt signaling in the dorsal otocyst determines semicircular canal formation in the mouse embryo

The mouse inner ear develops from a simple epithelial pouch, the otocyst, with the dorsal and ventral portions giving rise to the vestibule and cochlea, respectively. The otocyst undergoes a morphological change to generate flattened saclike structures, known as outpocketings, in the dorsal and lateral regions. The semicircular canals of the vestibule form from the periphery of the outpocketings, with the central region (the fusion plate) undergoing de-epithelialization and disappearing. However, little is known of the mechanism that orchestrates formation of the semicircular canals. We now show that the area of canonical Wnt signaling changes dynamically in the dorsal otocyst during its morphogenesis. The genes for several Wnt ligands were found to be expressed in the dorsal otocyst according to specific patterns, whereas those for secreted inhibitors of Wnt ligands were expressed exclusively in the ventral otocyst. With the use of whole-embryo culture in combination with potent modulators of canonical Wnt signaling, we found that forced persistence of such signaling resulted in impaired formation both of the lateral outpocketing and of the fusion plates of the dorsal outpocketing. Canonical Wnt signaling was found to suppress Netrin1 expression and to preserve the integrity of the outpocketing epithelium. In addition, inhibition of canonical Wnt signaling reduced the size of the otocyst, likely through suppression of cell proliferation and promotion of apoptosis. Our stage-specific functional analysis suggests that strict regulation of canonical Wnt signaling in the dorsal otocyst orchestrates the process of semicircular canal formation.

The mode of cerebellar activation of neurons in the cat motor cortex: an intracellular HRP study

Intracellular recordings and morphological identification of neurons by using intracellular HRP staining were performed in the cat motor cortex. By cerebellar stimulation, stellate cells in layers II-III, pyramidal cells in layer III and fast pyramidal tract neurons (PTNs) were activated with short latency and fast rising EPSPs, while pyramidal cells in layer II and slow PTNs showed a wide range of latency and slow rising EPSPs. This difference may be related to activation through the deep and superficial thalamocortical projections.

Inflammatory pseudotumor in head and neck

BACKGROUND Inflammatory pseudotumor (IPT) is a tumefactive lesion characterized by fibroblastic proliferations and a prominent inflammatory component. It behaves as a locally benign or aggressive lesion, clinically and radiologically mimicking a neoplastic process. Numerous entities can be diagnosed as IPT, from reactive lesions to true neoplasms. The diagnosis of IPT requires further elaboration, and IPT should be distinguished from other similar entities such as inflammatory myofibroblastic tumor and IgG4-related sclerosing disease. CASE SUMMARY We report two cases of IPT arising from the head and neck region. One occurred at the orbit and the other at the parapharyngeal space. Histologically, they showed aggregates of myofibroblasts and inflammatory cells. Immunohistochemically, the number of IgG4-positive cells was less than 40% of the number of IgG positive cells, and the myofibroblastic cells were negative for anaplastic lymphoma kinase. The diagnosis was IPT/not otherwise specified. One patient was treated by systemic administration of corticosteroid and had good response. The other, who was treated by local administration of corticosteroid, partially responded and is currently stable with limited disease. DISCUSSION IPT has been reported to occur in various anatomical sites, most commonly in the lungs. The incidence in the head and neck area is extremely rare. Treatment of IPT is controversial and may involve corticosteroids or surgical resection, or both. Other chemotherapeutic agents and radiotherapy may be considered in steroid-resistant patients. The pathological subtype, safety of resection, and safety of corticosteroid use must be included in the decision-making process for treatment.

Property of IK,n in inner hair cells isolated from guinea-pig cochlea

One of the potassium currents, I(K,)(n), is already activated at the resting potential of the cell and thus determines the membrane potential. KCNQ4 channel has been identified as the molecular correlate of I(K,)(n). In the present study, we measured I(K,)(n) in acutely isolated IHCs of guinea-pig cochlea using the whole-cell voltage-clamp techniques, and investigated the properties of the currents. I(K,)(n) was 70% activated around the resting potential of -60 mV and deactivated on hyperpolarization. I(K,)(n) was blocked by the KCNQ-channel blockers, linopirdine (100 microM) and XE991 (10 microM), but was insensitive to both I(K,f) blocker, tetraethylammonium (TEA), and I(K,s) blocker, 4-aminopyridine (4-AP). There was no significant difference in the size of I(K,)(n) between the apical and basal turn IHCs.

Salicylate-induced morphological changes of isolated inner hair cells and outer hair cells from guinea-pig cochlea

OBJECTIVES The aim of this study is to investigate the salicylate-induced morphological changes of cochlear inner hair cells (IHCs) and outer hair cells (OHCs). METHODS IHCs and OHCs were acutely isolated from the guinea-pig cochlea. Cells were observed under the inverted microscope and 10mmol/L sodium salicylate solutions or 0.01mmol/L dexamethasone-plus-salicylate solutions were applied. The cell length or the ratio between the length and width was the indices of the morphological changes in cells. RESULTS Isolated IHCs did not demonstrate any significant changes in sodium salicylate solutions in 20min and in 40min, whereas OHCs were shortened by the 10mmol/L sodium salicylate to 83% in 20min and 75% in 40min. There were no significant differences between in the dexamethasone-plus-salicylate solutions and in the control solutions after 20min and 40min both in IHCs and OHCs. CONCLUSIONS Although salicylate affected the isolated OHCs from guinea-pig cochlea, IHCs were not changed morphologically by sodium salicylate applications. Dexamethasone inhibited the salicylate-induced morphological changes of OHCs.

Effect of salicylate on potassium currents in inner hair cells isolated from guinea-pig cochlea

Although salicylate is one of the most widely used nonsteroidal anti-inflammatory drugs, it causes moderate hearing loss and tinnitus at high-dose levels. In the present study, salicylate effects on the K currents in inner hair cells were examined. Salicylate reversibly reduced the outward K currents (I(K,f)), but did not affect the inward current (I(K,n)). Salicylate blocked the outward K currents in a concentration-dependent manner according to Hill equation with a half-blocking concentration of 1.66mM, and the Hill coefficient of 1.86.

Direct reprogramming of spiral ganglion non-neuronal cells into neurons: Toward ameliorating sensorineural hearing loss by gene therapy

Primary auditory neurons (PANs) play a critical role in hearing by transmitting sound information from the inner ear to the brain. Their progressive degeneration is associated with excessive noise, disease and aging. The loss of PANs leads to permanent hearing impairment since they are incapable of regenerating. Spiral ganglion non-neuronal cells (SGNNCs), comprised mainly of glia, are resident within the modiolus and continue to survive after PAN loss. These attributes make SGNNCs an excellent target for replacing damaged PANs through cellular reprogramming. We used the neurogenic pioneer transcription factor Ascl1 and the auditory neuron differentiation factor NeuroD1 to reprogram SGNNCs into induced neurons (iNs). The overexpression of both Ascl1 and NeuroD1 in vitro generated iNs at high efficiency. Transcriptome analyses revealed that iNs displayed a transcriptome profile resembling that of endogenous PANs, including expression of several key markers of neuronal identity: Tubb3, Map2, Prph, Snap25, and Prox1. Pathway analyses indicated that essential pathways in neuronal growth and maturation were activated in cells upon neuronal induction. Furthermore, iNs extended projections toward cochlear hair cells and cochlear nucleus neurons when cultured with each respective tissue. Taken together, our study demonstrates that PAN-like neurons can be generated from endogenous SGNNCs. This work suggests that gene therapy can be a viable strategy to treat sensorineural hearing loss caused by degeneration of PANs.

Kremen1 regulates mechanosensory hair cell development in the mammalian cochlea and the zebrafish lateral line

Here we present spatio-temporal localization of Kremen1, a transmembrane receptor, in the mammalian cochlea, and investigate its role in the formation of sensory organs in mammal and fish model organisms. We show that Kremen1 is expressed in prosensory cells during cochlear development and in supporting cells of the adult mouse cochlea. Based on this expression pattern, we investigated whether Kremen1 functions to modulate cell fate decisions in the prosensory domain of the developing cochlea. We used gain and loss-of-function experiments to show that Kremen1 is sufficient to bias cells towards supporting cell fate, and is implicated in suppression of hair cell formation. In addition to our findings in the mouse cochlea, we examined the effects of over expression and loss of Kremen1 in the zebrafish lateral line. In agreement with our mouse data, we show that over expression of Kremen1 has a negative effect on the number of mechanosensory cells that form in the zebrafish neuromasts, and that fish lacking Kremen1 protein develop more hair cells per neuromast compared to wild type fish. Collectively, these data support an inhibitory role for Kremen1 in hair cell fate specification.

Cochlear implants for mumps deafness: two paediatric cases.

BACKGROUND Good outcomes have been reported regarding the use of cochlear implants for mumps deafness. The mumps virus induces meningitis and/or encephalitis, which can cause central nervous system damage resulting in retrolabyrinthine hearing loss, for which a cochlear implant would be less effective. CASES We installed a cochlear implant in two patients with bilateral mumps deafness; one achieved a good result with the cochlear implant, but the other did not. We discuss two possible reasons for the different outcomes. Case 1 was a three-year-old girl with bilateral parotid swelling, vomiting and walking disorder. One year after cochlear implant insertion, speech perception did not develop despite of good pure tone thresholds. Case 2 was an eight-year-old girl with bilateral parotid swelling. A cochlear implant enabled her to improve hearing perception. CONCLUSION Although cochlear implants have been reported to be helpful for mumps deafness, cases that involve central nervous system damage may not achieve good results.