Hajime Sawai

Pikachurin, a dystroglycan ligand, is essential for photoreceptor ribbon synapse formation

Exquisitely precise synapse formation is crucial for the mammalian CNS to function correctly. Retinal photoreceptors transfer information to bipolar and horizontal cells at a specialized synapse, the ribbon synapse. We identified pikachurin, an extracellular matrix–like retinal protein, and observed that it localized to the synaptic cleft in the photoreceptor ribbon synapse. Pikachurin null-mutant mice showed improper apposition of the bipolar cell dendritic tips to the photoreceptor ribbon synapses, resulting in alterations in synaptic signal transmission and visual function. Pikachurin colocalized with both dystrophin and dystroglycan at the ribbon synapses. Furthermore, we observed direct biochemical interactions between pikachurin and dystroglycan. Together, our results identify pikachurin as a dystroglycan-interacting protein and demonstrate that it has an essential role in the precise interactions between the photoreceptor ribbon synapse and the bipolar dendrites. This may also advance our understanding of the molecular mechanisms underlying the retinal electrophysiological abnormalities observed in muscular dystrophy patients.

Effects of Adiponectin on the Renal Sympathetic Nerve Activity and Blood Pressure in Rats

Adiponectin is an adipocytokine that modulates energy homeostasis and glucose metabolism. Here, we examined the effects of acute intravenous (iv) and lateral cerebral ventricular (LCV) injections of adiponectin on the renal sympathetic nerve activity (RSNA) and blood pressure (b/p) in urethane-anesthetized rats. Both iv and LCV injections of adiponectin induced dose-dependent suppressions of RSNA and b/p. Moreover, we found that bilateral lesions of the hypothalamic suprachiasmatic nucleus (SCN) abolished the effects of iv injection of adiponectin on RSNA and b/p. These findings suggest that adiponectin decreases the RSNA and b/p in a dose-dependent manner and that the SCN is implicated in mechanism of adiponectin actions on RSNA and b/p. These findings also suggest that the hypotensive-action activity of adiponectin is realized, at least partially, via changes in activities of autonomic nerves activity.

Axonal regeneration induced by repetitive electrical stimulation of crushed optic nerve in adult rats

PurposeTo investigate whether electrical stimulation promoted axonal regeneration of retinal ganglion cells (RGCs) after optic nerve (ON) crush in adult rats.MethodsTranscorneal electrical stimulation (TES), which stimulates the retina with current from a corneal contact lens electrode, was used to stimulate the eye. TES was applied for 1 h immediately after ON crush. Axonal regeneration was determined by anterograde labeling of RGC axons. To examine whether the axonal regeneration was mediated by insulin-like growth factor 1 (IGF-1) receptors, an IGF-1 receptor antagonist, JB3, was injected intraperitoneally before each TES application. Immunostaining for IGF-1 was performed to examine the effects of TES. To test the survival-promoting effects of TES applied daily, the mean density of retrogradely labeled RGCs was determined on day 12 after ON crush.ResultsCompared with sham stimulation, the mean number of regenerating axons significantly increased at 250 μm distal from the lesion and increased IGF-1 immunoreactivity was observed in retinas treated daily with TES. Preinjection of an IGF-1 receptor antagonist significantly blocked axonal regeneration by TES applied daily. TES applied daily also markedly enhanced the survival of RGCs 12 days after ON crush.ConclusionTES applied daily promotes both axonal regeneration and survival of RGCs after ON crush.

Optimal parameters of transcorneal electrical stimulation (TES) to be neuroprotective of axotomized RGCs in adult rats

We previously showed that transcorneal electrical stimulation (TES) promoted the survival of axotomized retinal ganglion cells (RGCs) of rats. However the relationship between the parameters of TES and the neuroprotective effect of TES on axotomized RGCs was unclear. In the present study, we determined whether the neuroprotective effect of TES is affected by the parameters of TES. Adult male Wistar rats received TES just after transection of the left optic nerve (ON). The pulse duration, current intensity, frequency, waveform, and numbers of sessions of the TES were changed systematically. The alterations of the retina were examined histologically seven days or fourteen days after the ON transection. The optimal neuroprotective parameters were pulse duration of 1 and 2 ms/phase (P < 0.001, each), current intensity of 100 and 200 muA (P < 0.05, each), and stimulation frequency of 1, 5, and 20 Hz (P < 0.001, respectively). More than 30 min of TES was necessary to have a neuroprotective effect (P < 0.001). Symmetric pulses without an inter-pulse interval were most effective (P < 0.001). Repeated TES was more neuroprotective than a single TES at 14 days after ON transection (P < 0.001). Our results indicate that there is a range of optimal neuroprotective parameters of TES for axotomized RGCs of rats. These values will provide a guideline for the use of TES in patients with different retinal and optic nerve diseases.

Regulation of autonomic nerve activities by central pituitary adenylate cyclase-activating polypeptide

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a peptidergic neurotransmitter that is expressed in high levels in nervous systems. Here, we investigated the roles of PACAP in autonomic system regulation by evaluating the changes caused in the autonomic nerve activities after injecting PACAP into the central nervous system (CNS) and examining stress-induced blood glucose changes in PACAP-deficient (PACAP-/-) mice. Renal sympathetic nerve activity (RSNA), blood pressure, and heart rate were elevated after injecting PACAP into the third cerebral ventricle (3CV). Similarly, other sympathetic nerve activities, including adrenal sympathetic nerve activity (ASNA), celiac sympathetic nerve activity (CSNA), and brown adipose tissue sympathetic nerve activity (BAT-SNA), were accelerated by PACAP injection. In contrast, injecting PACAP into 3CV significantly suppressed parasympathetic nerve activities, including gastric vagal nerve activity (GVNA) and celiac vagal nerve activity (CVNA). In addition, blood glucose elevations induced by stress, such as immobilization or ether exposure, were disrupted in PACAP-/- mice, although basal glucose levels in mutants were comparable to that in wild-type mice. These results suggest that CNS PACAP regulates autonomic function by maintaining a sympathetic-parasympathetic balance and contributes to peripheral homeostatic maintenance, especially under conditions of stress.

Axonal regeneration of retinal ganglion cells in the cat geniculocortical pathway

The optic nerve of anesthetized cats was completely cut and the autologous sciatic nerve was transplanted. Sixty days later some populations of retinal ganglion cells were shown to regenerate the axon with retrograde HRP labeling. We verified that ganglion cells that had projected to the lateral geniculate nucleus (LGN) were able to regenerate through the transplant with a double-labeling method: diI was injected into the LGN prior to the transplantation, and dextran-fluorescein was injected into the graft after axonal regeneration. Intracellular injection of HRP into regenerating ganglion cells in an in vitro preparation revealed that the two major cell types projecting to the LGN, alpha and beta, regenerated axons and showed normal dendritic morphology.

Dose-Different Effects of Orexin-A on the Renal Sympathetic Nerve and Blood Pressure in Urethane-Anesthetized Rats

Previous studies have demonstrated that central injection of orexin-A affects renal sympathetic nerve activity (RSNA) and blood pressure (BP) in both anesthetized and unanesthetized rats. In the present study, we examined, using urethane-anesthetized rats, the dose-dependent effects of intravenous (iv) or intralateral cerebral ventricular (LCV) injection of various doses of orexin-A on RSNA and BP. We found that injection of a low dose of orexin-A (10 ng iv or 0.01 ng LCV) suppressed RSNA and BP significantly. Conversely, a high dose (1000 ng iv or 10 ng LCV) of orexin-A elevated both RSNA and BP significantly. Pretreatment with either iv or LCV injection of thioperamide, a histaminergic H3-receptor antagonist, eliminated the effects of a low dose of orexin-A on both RSNA and BP. Both iv and LCV injection of diphenhydramine, a histaminergic H1-receptor antagonist, abolished the effects of a high dose of orexin-A on RSNA and BP. Furthermore, bilateral lesions of the hypothalamic suprachiasmatic nucleus (SCN) abolished the effects of both low and high doses of orexin-A on RSNA and BP. These findings suggest that orexin-A affects RSNA and BP in a dose-dependent manner and that the SCN and histaminergic nerve may be involved in the dose-different effects of orexin-A in rats.

Retinotopic organizations of the expanded ipsilateral projection to the rat's superior colliculus; variations along its rostrocaudal axis.

The retinotopic organization of the expanded projection to the ipsilateral colliculus was studied electrophysiologically in 12 adult albino rats with one eye removed at birth. For comparison, the contralateral projection was rather variable in the representation of the nasotemporal axis of the retina along the rostrocaudal dimension of the colliculus; on the other hand, the representation of dorsal-to-ventral retinal axis onto the mediolateral dimension of the colliculus was relatively stable.

Effects of EEG synchronization on visual responses of the cat's geniculate relay cells: a comparison among Y,X and W cells

Single unit activities of Y, X and W cells were recorded in the cat lateral geniculate nucleus to study how their responses to stationary light spots change with the cortical EEG. The shift from desynchronized to synchronized EEG drastically suppressed W cell activities, depressed both transient and sustained components of X cell responses, but did not affect the transient component of Y cell responses. This was ascribed to different inhibitory circuitries in the three parallel pathways and their differential modulation by the ascending brainstem activities.

Intraretinal axons of ganglion cells in the Japanese monkey (Macaca fuscata): conduction velocity and diameter distribution

In anesthetized and immobilized Japanese monkeys (Macaca fuscata), intraretinal conduction velocities of the ganglion cell axons were measured. The field potentials elicited by optic chiasm shocks consisted of fast and slow components with estimated conduction velocities of 1.19 and 0.72 m/s in recordings from the optic nerve fiber layer, and 1.65 and 1.00 m/s in recordings from the ganglion cell layer. Single cell recordings verified that the time course of the fast component corresponded to the antidromic spike latencies of Y-like cells, whereas that of the slow component covered the latency range of both X-like and W-like cells. In an electron microscopic study of the cross-sections of the intraretinal optic nerve fiber bundles, the axon diameter histograms of large samples (n = 3000-6000) all showed a unimodal distribution with a sharp peak at 0.3-0.6 micron and a long tail extending to 2-3 micron. The mean diameter was largest in the ventral and nasal bundles, smallest in the papillomacular bundle and intermediate in the dorsal, upper arcuate and lower arcuate bundles. However, diameter histograms of a small number of regional axons (n = 255-300) showed a broad tail distinct from the peak at 0.3-0.6 micron, enabling us to segregate a group of larger axons from the medium-sized to small axons. From such regional axon diameter histograms we estimated the mean relative occurrences of the larger axons (7.1-11.3%) and their mean diameters (0.9-1.3 micron). We further applied this relative frequency to the unimodal distribution of the histograms with larger samples in the upper and lower arcuate bundles and estimated the mean axon diameter of the large axons (1.1 micron) and that of the medium-sized to small axons (slightly below 0.5 micron). Finally, in studying the relation between axon diameter and conduction velocity in the two arcuate fiber bundles, we found it to be somewhat different from that previously reported for the cat retina.