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Studies on production of nuclear transplanted mouse embryos (핵이식 마우스 생산에 관한 연구)

  • Lee, Byeong-chun;Jo, Choong-ho;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.151-169
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    • 1993
  • The present study was carried out to investigate the best condition for nuclear-cytoplasm fusion and in vitro culture of nuclear transplanted embryos and to investigate the production of nuclear transplanted offsprings. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulses at output voltage of 1.0, 1.5 and 2.0 kV/cm for 100, 150 and $200{\mu}$ sec to induce cell fusion. 1. The culture of intact or zona cut 2-cell embryos in the medium supplemented with cytochalasin B($5{\mu}g/m{\ell}$) and colcemide($0.1{\mu}g/m{\ell}$)for 30 and 60 minutes did not affect the development to later stage. 2. The in vitro developmental rates of group A(a nucleus from one of the blastomeres was removed) and B(electrofusion of group A) were significantly lower than that of control group(p<0.01). 3. When nuclear transplanted embryos were submitted to electrofusion, the significantly higher fusion rates of 2-cell donor nuclei were achieved at the electric field strength of DC 1.5kV/cm for 100 and $150{\mu}$ sec, DC 2.0 kV/cm for $100{\sim}200{\mu}$ sec than DC 1.0 kV/cm for 100 and $150{\mu}$ sec(p<0.01). The significantly higher fusion rates of 4-cell donor nuclei were achieved at DC 2.0 kV/cm for 100 and $150{\mu}$ sec than DC 1.0kV/cm for $100{\sim}200{\mu}$ sec(p<0.01). These fusion rates in 8-cell donor nuclei were 88.7~99.3%. 4. The developmental potency to blastocyst in 2- and 4-cell donor nuclei was significantly higher in DC 1.0 and 2.0 kV/cm for $100{\sim}200{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group (p<0.01). The developmental potency to blastocyst in 8-cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{\mu}$ sec treated group than in DC 1.0 kV/cm for $100{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group(p<001). 5. The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 6. The success rate of nuclear injection into enucleated 2-cell embryos was significantly higher in 2-cell donor nuclei than in 4- or 8-cell donor nuclei(p<0.01). 7. The culture time taken for the nuclear transplanted 2-cell embryos to blastocyst stage was significantly longer in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 8. There was no significant difference in the developmental potency of nuclear transplanted embryos within the concentration of EGF at 0 to 15 ng per $m{\ell}$ of BMOC-3 solution. 9. The production rates of offspring after transfer of nuclear transplanted embryos to recipient mouse were significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01).

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Effect of $K^+-channel$ Blockers on the $A_1-adenosine$ Receptor-Coupled Regulation of Electrically-Evoked Norepinephrine Release in the Rat Hippocampus (흰쥐 해마에서 Norepinephrine 유리를 조절하는 $A_1-adenosine$ 수용체의 역할에 미치는 $K^+$ 통로 차단제의 영향)

  • Choi, Bong-Kyu;Kim, Sang-Hoon
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.301-309
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    • 1996
  • Since it has been reported that the depolarization-induced NE release is inhibited by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus, a large body of experimental data on the post-receptor mechanism of this process has been accumulated. But, the post-receptor mechanism of presynaptic $A_1-adenosine$ receptor on the NE release has not been clearly elucidated yet. Therefore, it was attempted to clarify the participation of $K^+-channel$ in the post-receptor mechanisms of the $A_1-adenosine$ receptor-mediated control of NE release in this study. Slices from rat hippocampus were equilibrated with $^3H-norepinephrine$ and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 $VCm^{-1}$, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Adenosine, in concentrations ranging from $1{\sim}30\;{\mu}M$, decreased the NE release in a dose-dependent manner, without affecting the basal rate of release. 4AP $(1{\sim}30{\mu}M)$, a specific A-type $K^+-channel$ blocker, increased the evoked NE release in a dose-related fashion, and the basal rate of release is increased by 10 and $30{\mu}M$. TEA $(1{\sim}10{\mu}\;M)$, a nonspecific $K^+-channel$ blocker, increased the evoked NE release in a dose-dependent manner without affecting basal release. The adenosine effects were significantly inhibites by 3 ${\mu}M$ 4AP and 10 mM TEA treatment. 4AP $(30{\mu}M)-$ and TEA (10 mM)-induced increments of evoked NE release were completely abolished in $Ca^{++} free, but these were recoverd in low $Ca^{++} medium. And the effects of $K^+-channel$ blockers in low $Ca^{++} medium were inhibites and abolishes by $Mg^{++} (4 mM) adding and TTX $(0.3{\mu}M)$ adding medium, respectively. These results suggest that the decrement of the evoked NE-release by $A_1-adenosine$ receptor is mediated by 4AP and TEA sensitive $K^+-channel$.

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The Role of Adenosine Receptors on Acetylcholine Release in the Rat Striatum

  • Kim, Do-Kyung;Kim, Hyeon-A;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.1-12
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    • 1997
  • As it has been reported that the depolarization induced acetylcholine (ACh) release is modulated by activation of presynaptic $A_1$ adenosine heteroreceptor and various evidence suggest that indicate the $A_2$ adenosine receptor is present in the striatum, this study was undertaken to delineate the role of adenosine receptors on the striatal ACh release. Slices from the rat striatum were equilibrated with $[^3H]$choline and then the release amount of the labelled product, $[^3H]$ACh, which was evoked by electrical stimulation (rectangular pulses, 3 Hz, 2 ms, 24 mA, $5\;Vcm^{-1}$, 2 min), was measured, and the influence of various agents on the evoked tritium outflow was investigated. And also, quantitative receptor autoradiography and drug-receptor binding assay were performed in order to confirm the presence and characteristics of $A_1$ and $A_2$ adenosine receptors in the rat striatum. Adenosine $(10{sim}100\;{mu}M)$ and $N^6$-cyclopentyladenosine (CPA, $1{sim}100\;{mu}M)$ decreased the $[^3H]$ACh release in a dose-dependent manner without changing the basal rate of release in the rat striatum. The reducing effects of ACh release by adenosine and CPA were abolished by 8-cyclopentyl-1,3-dipropy-Ixanthine (DPCPX, 2 ${mu}M$), a selective $A_1$, adenosine receptor antagonist, treatment. The effect of adenosine was potentiated markedly by 3,7-dimethyl-1-propargylxanthine (DMPX, 10 ${mu}M$), a specific $A_2$ adenosine receptor antagonist. 2-P-(2-carboxyethyl)phenethylamimo-5'-N- ethylcarboxamidoadenosine hydrochloride (CGS-21680C), in concentrations ranging from 0.01 to 10 ${mu}M$, a recently introduced potent $A_2$ adenosine receptor agonist, increased the $[^3H]$ACh release in a dose related fashion without changing the basal rate of release. These effects were completely abolished by DMPX $(10\;{mu}M)$. In autoradiograrhy experiments, $[^3H]$2-chloro-$N^6$-cyclopentyladenosine ($[^3H]$ CCPA) bindings were highly localized in the hippocampus and the cerebral cortex. Additionally, lower levels of binding were found in the striatum. However, $[^3H]$CGS-21680C bindings were highly localized in the striatal region with the greatest density of binding found in the caudate nucleus and putamen. Lower levels of binding were also found in the nucleus accumbens and olfactory tubercle. In drug-receptor binding assay, binding of $[^3H]$ CCPA to $A_1$ adenosine receptors of rat striatal membranes was inhibited by CPA ($K_i$ = 1.6 nM) and N-ethylcarboxamidoadenosine (NECA, $K_i$ = 12.9 nM), but not by CGS-21680C ($K_i$ = 2609.2 nM) and DMPX ($K_i$ = 19,386 nM). In contrast, $[^3H]$CGS-21680C binding to $A_2$ denosine receptors was inhibited by CGS-21680C ($K_i$ = 47.6 nM) and NECA ($K_i$ = 44.9 nM), but not by CPA ($K_i$ = 2099.2 nM) and DPCPX ($K_i$ = 19,207 nM). The results presented here suggest that both types of $A_1$ and $A_2$ adenosine heteroreceptors exist and play an important role in ACh release in the rat striatal cholinergic neurons.

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Monitoring on Radioactivity in Foodstuffs (식품에 대한 방사능 오염실태 조사)

  • Kwon, Ki-Sung;Hong, Jin-Hwan;Han, Sang-Bae;Lee, Eun-Ju;Kang, Kil-Jin;Chung, Hyung-Wook;Park, Seong-Gyu;Jang, Gui-Hyun;An, Ji-Seung;Kim, Dong-Sul;Kim, Myung-Chul;Kim, Chang-Min;Chung, Kun-Ho;Lee, Chang-Woo
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.183-187
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    • 2004
  • Radioactivity in foodstuffs was surveyed for reference in amending regulation on the maximum permitted levels of radioactive contamination of foodstuffs. Most domestic and imported (?) foodstuffs were sampled, some domestic items collected around nuclear power plants to compare site-specific contamination. The collected samples were dried and ashed. Radioactivity in foodstuffs was measured using HPGe gamma spectrometer, Cs-137 activity ranged from 0.025-0.053, 0.045-0.500, 0.062-0.105, 0.025-1.151, 0.021-0.145 and 0.046-0.155 Bq/kg-fresh in cereals, pulses, mot vegetables (potato), ginsengs, meat, and marine products, respectively, with imported dried ginseng showing the highest radioactivity, Results reveal radioactivity in foodstuffs collected in 2002 is far below the maximum permitted levels of 370 Bq/kg. No significant differences were observed in radioactivity among sampling sites and between domestic and imported foodstuffs.

A Study on the Post-Receptor Mechanism of Adenosine Receptor on Acetylcholine Release in the Rat Hippocampus (흰쥐 해마에서 Acetylcholine 유리에 관여하는 Adenosine Receptor의 Post-Receptor 기전에 관한 연구)

  • Choi, Bong-Kyu;Oh, Jae-Hee
    • The Korean Journal of Pharmacology
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    • v.30 no.3
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    • pp.263-272
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    • 1994
  • Since it was been reported that the depolarization-induced ACh release is inhibited by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus, a large body of experimental data on the post-receptor mechanism of this process has been accumulated. But, the post-receptor mechanism of presynaptic $A_1-adenosine$ receptor on the ACh release has not been clearly elucidated yet. Therefore, it was attempted to clarify the post-receptor mechanisms of the $A_1-adenosine$ receptor-mediated control of ACh release in this study. Slices from rat hippocampus were equilibrated with $^3H-choline$ and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 $VCm^{-1}$, 2ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Adenosine, in concentrations ranging from $0.3{\sim}300\;{\mu}M$, decreased the ACh release in a dose-dependent manner, without affecting the basal rate of release. The adenosine effects were significantly inhibited by $DPCPX\;(2\;{\mu}M)$, a selective $A_1-receptor$ antagonist. The responses to N-ethylmaleimide $(10&30{\mu}M)$, a SH-alkylating agent of G-protein, were characterized by increments of the evoked ACh-release and the basal release, and the adenosine effects were completely abolished by NEM pretreatment. PDB $(1{\sim}10\;{\mu}M)$, a specific protein kinase C (PKC) activator, increased, whereas PMB $(0.03{\sim}1\;mg)$, a PKC inhibitor, decreased the evoked ACh-release, and the adenosine effects were not affected by these agents. Nifedipine $(1\;{\mu}M)$, a $Ca^{2+}\;-channel$ blocker of dihydropyridine analogue, significantly inhibited the adenosine effect, but glibenclamide, a $K^+-channel$ blocker, did not. Finally, 8-bromo cyclic AMP $(100\;&\;300{\mu}M)$, a membrane-permeable analogue of cAMP, did not alter the ACh release, but adenosine effects were inhibited by pretreatment with large dose of 8-br-cAMP $(300\;{\mu}M)$. These results indicate that the decrement of the evoked ACh-release by $A_1-adenosine$ receptor is mediated by the G-protein, and nifedipine-sensitive $Ca^{2+}-channel$ and adenylate cyclase system are coupled partly to this effect, and that protein kinase C and glibenclamide-sensitive $K{^+}-channel$ are not involved in this process.

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Characteristics of the ( Pb, La ) $TiO_3$ Thin Films with Pb/La Compositions (Pb/La 조성에 따른 ( Pb, La ) $TiO_3$ 박막의 특성 변화)

  • Kang, Seong-Jun;Joung, Yang-Hee;Yoon, Yung-Sup
    • Journal of the Korean Institute of Telematics and Electronics D
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    • v.36D no.1
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    • pp.29-37
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    • 1999
  • In this study, we have prepared PLT thin films having various La concentrations by using sol-gel method and studied on the effect of La concentration on the electrical properties of PLT thin films. As the La concentration increases from 5mol% to 28mol%, the dielectric constant at 10kHz increases from 428 to 761, while the loss tangent decreases from 0.063 to 0.024. Also, the leakage current density at 150kV/cm has a tendency to decrease from 6.96${\mu}A/cm^2$ to 0.79${\mu}A/cm^2$. In the result of hysteresis loops of PLT thin films, the remanent polariation and the coercive field decrease from 9.55${\mu}C/cm^2$ to 1.10${\mu}C/cm^2$ and from 46.4kV/cm to 13.7kV/cm, respectively. With the result of the fatigue test on the PLT thin films, we have found that the fatigue properties are improved remarkably as the La concentration increases from 5 mol% to 28mol%. In particular, the PLT28) has paraelectric phase and its charge storage clensity and leakage current density at 5V are 134fC/${\mu}cm^2$ and 1.01${\mu}A/cm^2$, respectively. The remanent polarization and coercive field of the PLT(10) film are 6.96${\mu}C/cm^2$ and 40.2kV/cm, respectively. After applying of $10^9$ square pulses with ${\pm}5V$, the remanent polarilzation of the PLT(10) film decreases about 20% from the initial state. In the results, we conclude that the 10mol% and the 28mol% La doped PLT thin films are very suitable for the capacitor dielectrics of new generation of DRAM and NVFRAM respecitively.

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Validation of Food Intake Frequency from Food Frequency Questionnaire for Use as a Covariate in a Model to Estimate Usual Food Intake (식품의 일상섭취량 추정을 위한 식품섭취빈도의 활용가능성 및 타당도 연구)

  • Lee, Ja Yoon;Kim, Dong Woo
    • Culinary science and hospitality research
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    • v.23 no.2
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    • pp.64-73
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    • 2017
  • Although 24-hour recalls (24HR) capture detailed information on a person's food intake, this method suffers from difficulties in adequately measuring the usual intake of foods that are not consumed daily by most. Therefore, the purpose of this study is to investigate whether frequency of Food Frequency Questionnaire (FFQ) can be utilized in form of covariate when calculating usual intake of episodically-consumed foods and their distributions. Data used in this study was from the Korean National Healthy and Nutrition Examination Survey (KNHANES) 2012~2014 (3 years) and 10,945 subjects participated in this survey who performed both of 24HR and FFQ. In order to analyze the data, amount of intake in each food, which was reported in 24HR was recalculated according to 112 items in FFQ. We first assessed the relationship between FFQ frequency and the amount reported on 24HR. Second, we assessed the relationship between usual portion size of FFQ and the amount reported on 24HR. Our hypothesis was that people who reported high FFQ-reported frequency or FFQ-reported usual portion size would consume larger amounts of that food on 24HR than those with lower frequency or portion size of consumption of a food on the FFQ. For 59 of 112 individual foods (52.2%), there were statistically significant increasing relationships between FFQ frequency and consumption-day intake. Also, 102 of 112 individual foods (90.3%), there were statistically significant increasing relationships between FFQ usual portion size and consumption-day intake. For 10 of 13 food groups (grains, fruits, eggs, pulses, root and tuber crops, milk products, meat, beverage, alcoholic drink, vegetable, seaweeds and others), there were statistically significant increasing relationships between FFQ frequency and consumption-day intake. And there were statistically significant increasing relationships between FFQ usual portion size and consumption-day intake for all food groups. This study confirmed consistent correlation between reported FFQ frequency or usual portion size of food (group) consumption and consumption-day intake on 24HR. Therefore the frequency data may be utilized as important covariate when estimating usual intake of food or food groups.

A Study On Design of ZigBee Chip Communication Module for Remote Radiation Measurement (원격 방사선 측정을 위한 ZigBee 원칩형 통신 모듈 설계에 대한 연구)

  • Lee, Joo-Hyun;Lee, Seung-Ho
    • Journal of IKEEE
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    • v.18 no.4
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    • pp.552-558
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    • 2014
  • This paper suggests how to design a ZigBee-chip-based communication module to remotely measure radiation level. The suggested communication module consists of two control processors for the chip as generally required to configure a ZigBee system, and one chip module to configure a ZigBee RF device. The ZigBee-chip-based communication module for remote radiation measurement consists of a wireless communication controller; sensor and high-voltage generator; charger and power supply circuit; wired communication part; and RF circuit and antenna. The wireless communication controller is to control wireless communication for ZigBee and to measure radiation level remotely. The sensor and high-voltage generator generates 500 V in two consecutive series to amplify and filter pulses of radiation detected by G-M Tube. The charger and power supply circuit part is to charge lithium-ion battery and supply power to one-chip processors. The wired communication part serves as a RS-485/422 interface to enable USB interface and wired remote communication for interfacing with PC and debugging. RF circuit and antenna applies an RLC passive component for chip antenna to configure BALUN and antenna impedance matching circuit, allowing wireless communication. After configuring the ZigBee-chip-based communication module, tests were conducted to measure radiation level remotely: data were successfully transmitted in 10-meter and 100-meter distances, measuring radiation level in a remote condition. The communication module allows an environment where radiation level can be remotely measured in an economically beneficial way as it not only consumes less electricity but also costs less. By securing linearity of a radiation measuring device and by minimizing the device itself, it is possible to set up an environment where radiation can be measured in a reliable manner, and radiation level is monitored real-time.

A Study on nutritional status and dietary quality according to carbonated drink consumption in male adolescents: Based on 2007~2009 Korean National Health and Nutrition Examination Survey (남자 청소년의 탄산음료 섭취 빈도에 따른 영양소 섭취상태 및 식사의 질 평가: 2007~2009 국민건강영양조사 자료를 이용하여)

  • Bae, Yun-Jung;Yeon, Jee-Young
    • Journal of Nutrition and Health
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    • v.48 no.6
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    • pp.488-495
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    • 2015
  • Purpose: This study was conducted to evaluate nutrition intake and diet quality according to carbonated drink consumption in male adolescents (middle-school students = 480, high-school students = 417). Methods: We analyzed data from the combined 2007~2009 KNHANES (Korean National Health and Nutrition Examination Survey). Subjects were divided into two groups, the LCDI (low carbonated drink intake (< 1 time/week), n = 362) group and the HCDI (high carbonated drink intake (${\geq}1$ time/week), n = 535) group, according to carbonated beverage consumption. Nutrient and food group intake, NAR (nutrient adequacy ratio), and MAR (mean adequacy ratio) were analyzed using data from the 24-recall method. Results: Intake of plant protein, vitamin C, plant calcium, phosphorous, and potassium was significantly lower in the HCDI group, compared with the LCDI group. Percent of RNI (recommended nutrient intake) of vitamin C and phosphorous was significantly lower in the HCDI group, compared with the LCDI group. Percentage of subjects who consumed under EAR (estimated average requirement) of protein and vitamin C was significantly higher in the HCDI group, compared with the LCDI group. The NAR of phosphorous was significantly lower in the HCDI group, compared with the LCDI group. Food intakes from potato and starches, pulses and vegetables were significantly lower in the HCDI group, compared with the LCDI group. Conclusion: Consumption of carbonated drinks decreased the diet quality, including calcium, potassium, protein, and vitamin C. Therefore, nutrition education relating to consumption of carbonated drinks is required for male adolescents in order to maintain healthy dietary habits.

Comparison of Retinal Ganglion Cell Responses to Different Voltage Stimulation Parameters in Normal and rd1 Mouse Retina (정상망막과 변성망막에서 전압자극 파라미터 변화에 따른 망막신경절세포의 반응 비교)

  • Ye, Jang-Hee;Ryu, Sang-Baek;Kim, Kyung-Hwan;Goo, Yong-Sook
    • Progress in Medical Physics
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    • v.21 no.2
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    • pp.209-217
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    • 2010
  • Retinal prostheses are being developed to restore vision for the blind with retinal diseases such as retinitis pigmentosa (RP) or age-related macular degeneration (AMD). Since retinal prostheses depend upon electrical stimulation to control neural activity, optimal stimulation parameters for successful encoding of visual information are one of the most important requirements to enable visual perception. Therefore, in this paper, we focused on retinal ganglion cell (RGC) responses to different voltage stimulation parameters and compared threshold charge densities in normal and rd1 mice. For this purpose, we used in vitro preparation for the retina of normal and rd1 mice on micro-electrode arrays. When the neural network of rd1 mouse retinas is stimulated with voltage-controlled pulses, RGCs in degenerated retina also respond to voltage amplitude or voltage duration modulation as well in wild-type RGCs. But the temporal pattern of RGCs response is very different; in wild-type RGCs, single peak within 100 ms appears while in RGCs in degenerated retina multiple peaks (~4 peaks) with ~10 Hz rhythm within 400 ms appear. The thresholds for electrical activation of RGCs are overall more elevated in rd1 mouse retinas compared to wild-type mouse retinas: The thresholds for activation of RGCs in rd1 mouse retinas were on average two times higher ($70.50{\sim}99.87\;{\mu}C/cm^2$ vs. $37.23{\sim}61.65\;{\mu}C/cm^2$) in the experiment of voltage amplitude modulation and five times higher ($120.5{\sim}170.6\;{\mu}C/cm^2$ vs. $22.69{\sim}37.57\;{\mu}C/cm^2$) in the experiment of voltage duration modulation than those in wild-type mouse retinas. This is compatible with the findings from human studies that the currents required for evoking visual percepts in RP patients is much higher than those needed in healthy individuals. These results will be used as a guideline for optimal stimulation parameters for upcoming Korean-type retinal prosthesis.