• BMB Reports
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• 제42권7호
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• pp.456-461
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• 2009

Proteomic analyses of differentially expressed proteins in rat retinal ganglion cells (RGC-5) following S-nitrosoglutathione (GSNO), an NO donor, treatment were conducted. Of the approximately 314 protein spots that were detected, 19 were differentially expressed in response to treatment with GSNO. Of these, 14 proteins were up-regulated and 5 were down- regulated. Notably, an increase in GAPDH expression following GSNO treatment was detected in RGC-5 cells through Western blotting as well as proteomics. The increased GAPDH expression in response to GSNO treatment was accompanied by an increase in Herc6 protein, an E3 ubiquitin ligase. Moreover, GSNO treatment resulted in the translocation of GADPH from the cytosol to the nucleus and its subsequent accumulation. These results suggest that NO stress-induced apoptosis may be associated with the nuclear translocation and accumulation of GAPDH in RGC-5 cells.

• The Korean Journal of Physiology and Pharmacology
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• 제27권6호
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• pp.541-553
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• 2023

Retinal prostheses have shown some clinical success in restoring vision in patients with retinitis pigmentosa. However, the post-implantation visual acuity does not exceed that of legal blindness. The reason for the poor visual acuity might be that (1) degenerate retinal ganglion cells (RGCs) are less responsive to electrical stimulation than normal RGCs, and (2) electrically-evoked RGC spikes show a more widespread not focal response. The single-biphasic pulse electrical stimulation, commonly used in artificial vision, has limitations in addressing these issues. In this study, we propose the benefit of multiple consecutive-biphasic pulse stimulation. We used C57BL/6J mice and C3H/HeJ (rd1) mice for the normal retina and retinal degeneration model. An 8 × 8 multi-electrode array was used to record electrically-evoked RGC spikes. We compared RGC responses when increasing the amplitude of a single biphasic pulse versus increasing the number of consecutive biphasic pulses at the same stimulus charge. Increasing the amplitude of a single biphasic pulse induced more RGC spike firing while the spatial resolution of RGC populations decreased. For multiple consecutive-biphasic pulse stimulation, RGC firing increased as the number of pulses increased, and the spatial resolution of RGC populations was well preserved even up to 5 pulses. Multiple consecutive-biphasic pulse stimulation using two or three pulses in degenerate retinas induced as much RGC spike firing as in normal retinas. These findings suggest that the newly proposed multiple consecutive-biphasic pulse stimulation can improve the visual acuity in prosthesis-implanted patients.

• The Korean Journal of Physiology and Pharmacology
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• 제15권6호
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• pp.415-422
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• 2011

Previously, we reported that besides retinal ganglion cell (RGC) spike, there is ~10 Hz oscillatory rhythmic activity in local field potential (LFP) in retinal degeneration model, rd1 mice. The more recently identified rd10 mice have a later onset and slower rate of photoreceptor degeneration than the rd1 mice, providing more therapeutic potential. In this study, before adapting rd10 mice as a new animal model for our electrical stimulation study, we investigated electrical characteristics of rd10 mice. From the raw waveform of recording using $8{\times}8$ microelectrode array (MEA) from in vitro-whole mount retina, RGC spikes and LFP were isolated by using different filter setting. Fourier transform was performed for detection of frequency of bursting RGC spikes and oscillatory field potential (OFP). In rd1 mice, ~10 Hz rhythmic burst of spontaneous RGC spikes is always phase-locked with the OFP and this phase-locking property is preserved regardless of postnatal ages. However, in rd10 mice, there is a strong phase-locking tendency between the spectral peak of bursting RGC spikes (~5 Hz) and the first peak of OFP (~5 Hz) across different age groups. But this phase-locking property is not robust as in rd1 retina, but maintains for a few seconds. Since rd1 and rd10 retina show phase-locking property at different frequency (~10 Hz vs. ~5 Hz), we expect different response patterns to electrical stimulus between rd1 and rd10 retina. Therefore, to extract optimal stimulation parameters in rd10 retina, first we might define selection criteria for responding rd10 ganglion cells to electrical stimulus.

• The Korean Journal of Physiology and Pharmacology
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• 제21권5호
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• pp.555-563
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• 2017

Electrical stimulation through retinal prosthesis elicits both short and long-latency retinal ganglion cell (RGC) spikes. Because the short-latency RGC spike is usually obscured by electrical stimulus artifact, it is very important to isolate spike from stimulus artifact. Previously, we showed that topographic prominence (TP) discriminator based algorithm is valid and useful for artifact subtraction. In this study, we compared the performance of forward backward (FB) filter only vs. TP-adopted FB filter for artifact subtraction. From the extracted retinae of rd1 mice, we recorded RGC spikes with $8{\times}8$ multielectrode array (MEA). The recorded signals were classified into four groups by distances between the stimulation and recording electrodes on MEA (200-400, 400-600, 600-800, $800-1000{\mu}m$). Fifty cathodic phase-$1^{st}$ biphasic current pulses (duration $500{\mu}s$, intensity 5, 10, 20, 30, 40, 50, $60{\mu}A$) were applied at every 1 sec. We compared false positive error and false negative error in FB filter and TP-adopted FB filter. By implementing TP-adopted FB filter, short-latency spike can be detected better regarding sensitivity and specificity for detecting spikes regardless of the strength of stimulus and the distance between stimulus and recording electrodes.

• 한국식품영양과학회지
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• 제46권2호
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• pp.161-168
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• 2017

본 연구에서는 산화적 스트레스로 유도한 시신경 세포사멸에 대한 차조기 물 추출물(PFE)의 효과를 확인하였다. Staurosporine으로 분화된 ssdRGC-5 세포에 buthionine과 glutamate(B/G)로 산화적 스트레스를 유도하였으며, LDH release assay, MTT reduction assay를 통하여 PFE가 농도 의존적으로 B/G에 의한 세포사멸을 억제함을 관찰하였다. 세포사멸의 기전을 연구하기 위해 caspase 활성, 세포 내 ROS 생성량, 세포고사 관련 단백질 발현을 관찰한 결과, B/G에 의해 증가한 ROS 생성량, caspase 활성을 PFE가 억제하였고, 세포질로 방출된 cytochrome c와 미토콘드리아로 이동한 Bax도 감소함을 확인하였다. 이상의 결과로부터 차조기는 산화적 스트레스로 유도된 시신경 세포사멸 과정에서 항산화 효과와 미토콘드리아성 세포사멸을 완화함으로써 세포 보호 작용을 나타냄을 확인하였다.

• 대한의용생체공학회:의공학회지
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• 제28권2호
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• pp.212-217
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• 2007

As a preliminary study for the development of electrical stimulation strategy of artificial retina, we set up a method fur the reconstruction of input intensity variation from retinal ganglion cell(RGC) responses. In order to estimate light intensity variation, we used an optimal linear filter trained from given stimulus intensity variation and multiple single unit spike trains from RGCs. By applying ON/OFF stimulation(ON duration: 2 sec, OFF duration: 5 sec) repetitively, we identified three functional types of ganglion cells according to when they respond to the ON/OFF stimulus actively: ON cell, OFF cell, and ON-OFF cell. Experiments were also performed using a Gaussian random stimulus and a binary random stimulus. The input intensity was updated once every 90 msec(i. e. 11 Hz) to present the stimulus. The result of reconstructing 11 Hz Gaussian and binary random stimulus was not satisfactory and showed low correlation between the original and reconstructed stimulus. In the case of ON/OFF stimulus in which temporal variation is slow, successful reconstruction was achieved and the correlation coefficient was as high as 0.8.

• BMB Reports
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• 제50권2호
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• pp.60-70
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• 2017

Glaucoma is characterized by a slow and progressive degeneration of the optic nerve, including retinal ganglion cell (RGC) axons in the optic nerve head (ONH), leading to visual impairment. Despite its high prevalence, the biological basis of glaucoma pathogenesis still is not yet fully understood, and the factors contributing to its progression are currently not well characterized. Intraocular pressure (IOP) is the only modifiable risk factor, and reduction of IOP is the standard treatment for glaucoma. However, lowering IOP itself is not always effective for preserving visual function in patients with primary open-angle glaucoma. The second messenger cyclic adenosine 3',5'-monophosphate (cAMP) regulates numerous biological processes in the central nervous system including the retina and the optic nerve. Although recent studies revealed that cAMP generated by adenylyl cyclases (ACs) is important in regulating aqueous humor dynamics in ocular tissues, such as the ciliary body and trabecular meshwork, as well as cell death and growth in the retina and optic nerve, the functional role and significance of cAMP in glaucoma remain to be elucidated. In this review, we will discuss the functional role of cAMP in aqueous humor dynamics and IOP regulation, and review the current medications, which are related to the cAMP signaling pathway, for glaucoma treatment. Also, we will further focus on cAMP signaling in RGC growth and regeneration by soluble AC as well as ONH astrocytes by transmembrane ACs to understand its potential role in the pathogenesis of glaucoma neurodegeneration.

• 한국의학물리학회지:의학물리
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• 제21권2호
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• pp.209-217
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• 2010

색소성망막염(retinitis pigmentosa: RP)이나 연령관련 황반변성(age-related macular degeneration: AMD)과 같은 망막질환으로 인해 실명한 환자를 위해 인공시각장치가 개발되고 있다. 인공시각장치의 동작원리는 전기자극을 주어 신경세포의 활동도를 조절하는 것이므로 시각정보를 제대로 인코딩하기 위해 최적의 전기자극을 인가하는 것은 인공시각장치의 실용화를 위해 매우 중요한 요소이다. 그러므로 본 연구에서는 전압자극의 크기와 자극시간을 변화시켜 가면서 정상망막과 변성망막에 인가한 후 자극에 의해 유발된 망막신경절세포 반응을 분석하고 역치전하밀도를 비교함으로써 최적의 전기자극 조건을 찾아보고자 하였다. 이를 위하여 정상마우스와 rd1 마우스의 망막을 in vitro 상태로 분리한 후 망막의 신경절세포층이 전극을 향하여 부착되도록 한 후 망막신호를 기록하였다. rd1 마우스에서 얻은 변성망막의 망막신경절세포에서도 전압펄스를 인가시 정상망막의 망막신경절세포처럼 전압자극의 크기와 자극시간 변조에 대하여 반응하였다. 그러나 정상망막과 변성망막에서 망막신경절세포 반응의 시간적 패턴은 매우 달랐다: 정상망막의 망막신경절세포 반응은 전기자극 후 약 100 ms 내에서 1개의 피크만 나타나는 반면, 변성망막에서는 이보다 긴 400 ms 구간에서 약 10 Hz의 진동리듬을 가진 다수의 피크(~4개)들이 나타나는 것을 확인하였다. 또한 변성망막에서 망막신경절세포의 반응을 유발하기 위한 역치 전하밀도가 정상망막에서 보다 크게 상승하였다: 자극세기를 변화시켰을 때 정상망막의 역치 전하밀도는 $37.23{\sim}61.65\;{\mu}C/cm^2$, rd1 마우스에서는 $70.50{\sim}99.87\;{\mu}C/cm^2$로 2배가량 높은 것을 확인하였다. 자극시간을 변화시켰을 때 정상망막의 역치 전하밀도는 $22.69{\sim}37.57\;{\mu}C/cm^2$, rd1 마우스에서는 $120.5{\sim}170.6\;{\mu}C/cm^2$로 5배가량 높은 것을 확인하였다.