國內詞育 원숭이의 血淸 LDH의 總活性値와 isoenzyme에 관한 硏究

윤상보,김덕환,서지민,신남식,현병화,김명철,윤효인,박배근,송희종 한국임상수의학회 2001 한국임상수의학회지 Vol.18 No.4

Non-human primates have been increasing in demand as important experimental animals and companion animals, domestically and internationally. The number of non-human primates for these purposes will be much enhanced in the near future. Despite this trend, basic physiological data are scarcely available in these animal species, leading to the difficulty to diagnose diseases when necessary, due to the absence of reference values. Particularly, there is not any report on the total activity of LDH of non-human primates, let alone LDH isoenzyme patterns, in Korea. LDH isoenzymes have a high level of efficaciousness as diagnostic and prognostic aids in various diseases. In this study, total activities and isoenzyme patterns of LDH were measured to obtain their reference values in domestically reared common marmosets, crab-eating macaques and Japanese macaques. There were widespread different values of serum total LDH among the non-human primate species experimented in this study. Serum LDH values of common marmosets and crab-eating macaques were 597.5$\pm$243.1 IU/l and 605.3$\pm$312.6 IU/l, respectively, whereas those of Japanese macaque showed 1,209$\pm$473.8 IU/l. Five isoenzyme fractions of LDH were observed in all experimented non-human primates but their ranks and proportions represented different patterns one another. In common marmosets, the percent of fraction for serum LDH1, LDH$_2$, LDH$_3$, LDH$_4$, and LDH$_{5}$ was 13.7$\pm$6.4%, 23.3$\pm$3.6%, 29.2$\pm$5.0%, 9.4$\pm$1.4% and 24.4$\pm$7.5%, respectively. The rank of LDH isoenzymes was LDH$_3$>LDH$_{5}$>LDH$_2$>LDH$_1$>LDH$_4$, in the descending order. For crab-eating macaques, the fraction of serum LDH$_1$, LDH$_2$, LDH$_3$, LDH$_4$, and LDH$_{5}$ occupied 19.5$\pm$12.7%, 25.3$\pm$9.3%, 23.8$\pm$8.1%, 10.2$\pm$2.8% and 21.3$\pm$14.2%, respectively. The order of LDH isoenzymes was LDH$_2$>LDH$_3$>LDH$_{5}$>LDH$_1$>LDH$_4$, from top to down. On the while, in Japanese macaques, the fraction of serum LDH$_1$ to LDH$_{5}$ showed 23.4$\pm$11.8%, 30.5$\pm$4.1%, 17.4$\pm$3.9%, 11.3$\pm$3.7% and 13.8$\pm$5.6%, respectively. The decreasing order indicated LDH$_2$>LDH$_1$>LDH$_3$>LDH$_{5}$>LDH$_4$. In conclusion, values such as LDH and LDH isoenzyme patterns of investigated for the first time from non-human primates reaared in Korea, could be reference values for the optimal diagnosis and therapy of diseases of the corresponding animal species. Other parameters of hematology and blood biochemistry are urgently needed to study for the benefit of our intimate non-human primates.an primates.

Establishment of a Research and Assessment System Using High Quality Non-Human Primates

Byung-Hwa Hyun,Byeong-Cheol Kang,Chuel-Kyu Kim,Heui-Soo Kim,Jun Gyo Suh,Fumiaki Cho 한국실험동물학회 2010 Laboratory Animal Research Vol.26 No.4

At present, eight non-human primate research facilities exist in Korea to examine the validity and safety of new bio-products, and to generate model animal systems using primates of low health status (low quality primates). However specific-pathogen free (SPF) primates (high quality primates) are the preferred choice for emerging disease studies and for numerous other research areas, including cell/gene therapy, stem cell research, regenerative studies, and brain science. Although international primate centers in developed countries have utilized high quality primate resources for many years, there has been little or no collaboration with less developed countries on primate research. Due to this, the establishment of a high quality primate research capacity is a core priority for the advancement of the biomedical research field in less developed countries. In this study, we investigated the demand for, and opportunities to support the development of this research capability.

Abnormal Mitochondria in a Non-human Primate Model of MPTP-induced Parkinson’s Disease: Drp1 and CDK5/p25 Signaling

박준형,서진철,원진영,Hyeon-Gu Yeo,안유진,Keonwoo Kim,진영배,구본상,임경섭,Kang-Jin Jeong,Philyong Kang,Hwal-Yong Lee,Seung Ho Baek,Chang-Yeop Jeon,홍정주,허재원,김영현,Sang-Je Park,김선욱,이동석,이상래,이영전 한국뇌신경과학회 2019 Experimental Neurobiology Vol.28 No.3

Mitochondria continuously fuse and divide to maintain homeostasis. An impairment in the balance between the fusion and fission processes can trigger mitochondrial dysfunction. Accumulating evidence suggests that mitochondrial dysfunction is related to neurodegenerative diseases such as Parkinson’s disease (PD), with excessive mitochondrial fission in dopaminergic neurons being one of the pathological mechanisms of PD. Here, we investigated the balance between mitochondrial fusion and fission in the substantia nigra of a non-human primate model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD. We found that MPTP induced shorter and abnormally distributed mitochondria. This phenomenon was accompanied by the activation of dynamin-related protein 1 (Drp1), a mitochondrial fission protein, through increased phosphorylation at S616. Thereafter, we assessed for activation of the components of the cyclin-dependent kinase 5 (CDK5) and extracellular signal-regulated kinase (ERK) signaling cascades, which are known regulators of Drp1(S616) phosphorylation. MPTP induced an increase in p25 and p35, which are required for CDK5 activation. Together, these findings suggest that the phosphorylation of Drp1(S616) by CDK5 is involved in mitochondrial fission in the substantia nigra of a non-human primate model of MPTP-induced PD.

Identification and characterization of the tyrosinase gene (TYR) and its transcript variants (TYR_1 and TYR_2) in the crab-eating macaque (Macaca fascicularis)

Kim, Y.H.,Park, S.J.,Choe, S.H.,Lee, J.R.,Cho, H.M.,Kim, S.U.,Kim, J.S.,Sim, B.W.,Song, B.S.,Lee, Y.,Jin, Y.B.,Hong, J.J.,Jeong, K.J.,Kang, P.,Baek, S.H.,Lee, S.R.,Huh, J.W.,Chang, K.T. Elsevier/North-Holland 2017 Gene Vol.630 No.-

Tyrosinase is a copper-containing enzyme that regulates melanin biosynthesis and is encoded by the tyrosinase (TYR) gene. Previous studies demonstrated that mutations in TYR could lead to oculocutaneous albinism type 1 (OCA1) owing to the failure of melanin formation. Although a previous study found that albinism in the rhesus monkey was derived from a mutation in TYR, the identification and characterization of this gene in non-human primates has not been achieved thus far. Thus, using the rapid amplification of cDNA ends (RACE) and internal reverse transcription PCR (RT-PCR) we identified the full-length sequence of TYR in the crab-eating macaque, and two different transcript variants (TYR_1 and TYR_2). While TYR_1 comprised five exons and its coding sequence was highly similar to that of humans, TYR_2 comprised four exons and was generated by a third-exon-skipping event. Interestingly, these two transcripts were also present in the African green monkey (Old World monkey) and the common marmoset (New World monkey). Deduced amino acid sequence analyses revealed that TYR_2 had a shorter C-terminal region than TYR_1 owing to the exon-skipping event. Thus, the present study is the first to identify and characterize a full-length TYR gene in a non-human primate, while the further validation of the third-exon-skipping in TYR indicates that this event is well conserved in the primate lineage. Therefore, this study provides useful and important information for the study of albinism using non-human primate models.

Milk Transfer and Toxicokinetics of Valproic Acid in Lactating Cynomolgus Monkeys

Lee, Jong-Hwa,Yu, Wook-Joon,Jeong, Eun Ju,Chung, Moon-Koo Korean Society of ToxicologyKorea Environmental Mu 2013 Toxicological Research Vol.29 No.1

Studies on milk transfer of drugs in non-human primates (NHPs) are among the crucial components in the assessment of peri- and postnatal toxicity because of the similarity between NHPs and humans. To evaluate the milk transfer of valproic acid (VPA) in NHPs, the toxicokinetics of VPA, an antiepileptic drug, were studied in pregnant cynomolgus monkeys. VPA was administered once daily to pregnant cynomolgus monkeys at doses of 0, 30, 90, and 270 mg/kg by oral gavage from Day 100 of gestation (GD 100) to Day 31 of lactation (LD 31). Concentrations of VPA and its metabolite, 4-ene-VPA, in the maternal plasma on GD 100, GD 140, and LD 30, and concentrations of VPA and 4-ene-VPA in the offspring plasma and milk on LDs 30 and 31, respectively, were quantified using liquid chromatography tandem mass spectrometry (LC/MS/MS). After administration of a single oral dose of VPA to pregnant monkeys on GD 100, the concentrations of VPA and 4-ene-VPA were generally quantifiable in the plasma of all treatment groups up to 24 hr after administration, which showed that VPA was absorbed and that the monkeys were systemically exposed to VPA and 4-ene-VPA. After administration of multiple doses of VPA to the monkeys, VPA was detected in the pup's plasma and in milk taken on LD 30 and LD 31, respectively, which showed that VPA was transferred via milk, and the pup was exposed to VPA. Further, the concentration of VPA in the milk increased with an increase in the dose. Extremely low concentrations of 4-ene VPA were detected in the milk and in the pup plasma. In conclusion, pregnant monkeys were exposed to VPA and 4-ene-VPA after oral administration of VPA at doses of 30, 90, and 270 mg/kg/day from GD 100 to LD 31. VPA was transferred via milk, and the VPA exposure to the pup increased with an increase in the dose of VPA. The metabolite, 4-ene VPA, was present in extremely low concentrations (< 0.5 ${\mu}g/ml$) in the milk and in the pup plasma. In this study, we established methods to confirm milk transfer in NHPs, such as mating and diagnosis of pregnancy by examining gestational sac with ultrasonography, collection of milk and pup plasma and determination of toxicokinetics, using cynomolgus monkeys.

Milk Transfer and Toxicokinetics of Valproic Acid in Lactating Cynomolgus Monkeys

Jong-Hwa Lee,Wook-Joon Yu,Eun Ju Jeong,Moon-Koo Chung 한국독성학회 2013 Toxicological Research Vol.29 No.1

Studies on milk transfer of drugs in non-human primates (NHPs) are among the crucial components in the assessment of peri- and postnatal toxicity because of the similarity between NHPs and humans. To evaluate the milk transfer of valproic acid (VPA) in NHPs, the toxicokinetics of VPA, an antiepileptic drug, were studied in pregnant cynomolgus monkeys. VPA was administered once daily to pregnant cynomolgus monkeys at doses of 0, 30, 90, and 270 mg/kg by oral gavage from Day 100 of gestation (GD 100) to Day 31 of lactation (LD 31). Concentrations of VPA and its metabolite, 4-ene-VPA, in the maternal plasma on GD 100, GD 140, and LD 30, and concentrations of VPA and 4-ene-VPA in the offspring plasma and milk on LDs 30 and 31, respectively, were quantified using liquid chromatography tandem mass spectrometry (LC/MS/MS). After administration of a single oral dose of VPA to pregnant monkeys on GD 100, the concentrations of VPA and 4-ene-VPA were generally quantifiable in the plasma of all treatment groups up to 24 hr after administration, which showed that VPA was absorbed and that the monkeys were systemically exposed to VPA and 4-ene-VPA. After administration of multiple doses of VPA to the monkeys, VPA was detected in the pup’s plasma and in milk taken on LD 30 and LD 31, respectively, which showed that VPA was transferred via milk, and the pup was exposed to VPA. Further, the concentration of VPA in the milk increased with an increase in the dose. Extremely low concentrations of 4-ene VPA were detected in the milk and in the pup plasma. In conclusion, pregnant monkeys were exposed to VPA and 4-ene-VPA after oral administration of VPA at doses of 30, 90, and 270 mg/kg/day from GD 100 to LD 31. VPA was transferred via milk, and the VPA exposure to the pup increased with an increase in the dose of VPA. The metabolite, 4-ene VPA, was present in extremely low concentrations (< 0.5 μg/ml) in the milk and in the pup plasma. In this study, we established methods to confirm milk transfer in NHPs, such as mating and diagnosis of pregnancy by examining gestational sac with ultrasonography, collection of milk and pup plasma and determination of toxicokinetics, using cynomolgus monkeys.

Non-human primate using and researches as a country without primate resources

Byung Hwa Hyun 한국실험동물학회 2015 한국실험동물학회 학술발표대회 논문집 Vol.2015 No.8

Simplified adaptor for stereotactic surgery in non-human primates

Kim, Hyung-Sun,Byun, Donghak,Kim, Ra Gyung,Kang, Goo-Hwa,Park, Ji-Young,Yang, Young-Su,Han, Su-Cheol,Kim, Hyoung-Ihl Elsevier 2018 Journal of neuroscience methods Vol.295 No.-

<P><B>Abstract</B></P> <P><B>Background</B></P> <P>It is challenging for researchers performing stereotactic procedures to transition from small animals to non-human primate (NHP) experiments. The NHP stereotactic atlas is based on ear-bar zero (EBZ), which is an anatomical reference frame that is not visible during surgery. Most current NHP stereotactic systems require high-cost MRI or CT imaging and complex computer processing to determine the stereotactic coordinates, limiting the procedure to those with significant expertise.</P> <P><B>New method</B></P> <P>We have designed a simplified adaptor consisting of a circular arc for coronal tilt, a carrier for electrodes or cannulas, and an anchor to attach the adaptor to a conventional stereotactic frame. Our adaptor allows easy identification of the EBZ with the help of an anchor notch, and provides digital distance sensors without the need for imaging data or computer processing. Our system enables the use of trajectories that avoid injury to important structures and vessels.</P> <P><B>Results</B></P> <P>We tested the accuracy of our system using simulated targeting with phantoms, and demonstrated sub-millimeter accuracy. Infusion of methylene blue also showed satisfactory staining in target structures deep in the brain.</P> <P><B>Comparison with existing methods</B></P> <P>This system does not require high-cost imaging and extra training to determine EBZ. Once EBZ is set automatically by the system itself, targeting is similar to that in small animal stereotactic procedure.</P> <P><B>Conclusion</B></P> <P>Our simple adaptor will aid researchers who plan to conduct experiments involving stereotactic surgery in NHPs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A simple adaptor is proposed for stereotactic surgery in non-human primates. </LI> <LI> This adaptor can be attached to a conventional animal stereotactic frame. </LI> <LI> It allows identification of ear bar zero without imaging and computer processing. </LI> <LI> Simulation study shows the submillimeter precision in targeting. </LI> <LI> The proposed system will be useful for unexperienced translational researcher. </LI> </UL> </P>

Non-human primate diagnostic testing services of K-MEDI hub preclinical research center

Minji Seong,Se-Kyung Oh,Hyejin Kim,Dongkyu Kim,Kil-Soo Kim,Eui-Suk Jeong 한국실험동물학회 2022 한국실험동물학회 학술발표대회 논문집 Vol.2022 No.7

Expression and localization of fluorescent gene induced by intravitreal delivery in non-human primate retina: histological analysis

Jincheol Seo,Lee Wha Gwon,Jongmin Kim,Kyung Seob Lim,Chang-Yeop Jeon,Dong-Woo Cho,Seung Hwan Lee,Jae Yon Won,Youngjeon Lee 한국실험동물학회 2023 한국실험동물학회 학술발표대회 논문집 Vol.2023 No.2