• Molecules and Cells
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• 제19권3호
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• pp.408-417
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• 2005

Nitric oxide (NO) occurs in various types of cells in the central nervous system. We studied the distribution and morphology of neuronal nitric oxide synthase (NOS)-containing neurons in the visual cortex of mouse and rabbit with antibody immunocytochemistry. We also compared this labeling to that of calbindin D28K, calretinin, and parvalbumin. Staining for NOS was seen both in the specific layers and in selective cell types. The densest concentration of intense anti-NOS immunoreactive (IR) neurons was found in layer VI, while the weak anti-NOS-IR neurons were found in layer II/III in both animals. The NOS-IR neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. Two-color immunofluorescence revealed that only 16.7% of the NOS-IR cells were double-labeled with calbindin D28K in the mouse visual cortex, while more than half (51.7%) of the NOS-IR cells were double-labeled with calretinin and 25.0% of the NOS-IR cells were double-labeled with parvalbumin in mouse. By contrast, 92.4% of the NOS-IR neurons expressed calbindin D28K while only 2.5% of the NOS-IR neurons expressed calretinin in the rabbit visual cortex. In contrast with the mouse, none of the NOS-IR cells in the rabbit visual cortex were double-labeled with parvalbumin. The results indicate that neurons in the visual cortex of both animals express NOS in specific layers and cell types, which do not correlate with the expression of calbindin D28K, calretinin or parvalbumin between the two animals.

• 한국양식학회지
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• 제16권1호
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• pp.8-14
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• 2003

To generate expressed sequence tags for genomics research involving genetic linkage analysis, to examine gene expression profiles in muscles of channel catfish in a non-normalized muscle cDNA library, to compare gene expression in young and mature channel catfish muscles using the EST reagents and gene filters to demonstrate the feasibility of functional genomics research in small laboratories. 102 randomly picked cDNA clones were analyzed from the catfish muscle cDNA library. Of the sequences generated, 90.2% of ESTs was identified as known genes by identity comparisons. These 92 clones of known gene products represent transcriptional products of 24 genes. The 10 clones of unknown gene products represent 8 genes. The major transcripts (70.1% of the analyzed ESTs) in the catfish muscle are from many major genes involved in muscle contraction, relaxation, energy metabolism and calcium binding such as alpha actin, creatine kinase, parvalbumin, myosin, troponins, and tropomyosins. Gene expression of the unique ESTs was comparatively studied in the young and adult catfish muscles. Significant differences were observed for aldolase, myostatin, myosin light chain, parvalbumin, and an unknown gene. While myosin light chain and an unknown gene (CM 192) are down-regulated in the mature fish muscle, the aldolase, myostatin, and parvalbumin are significantly up-regulated in the mature fish muscle. Although the physiological significance of the changes in expression levels needs to be further addressed, this research demonstrates the feasibility and power of functional genomics in channel catfish. Channel catfish muscle gene expression profiles provide a valuable molecular muscle physiology blueprint for functional comparative genomics.

• Molecules and Cells
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• 제21권1호
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• pp.34-41
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• 2006

The neuronal localization of alpha-amino-3-hydroxyl-5-methyl-4-isoxazole propionic acid (AMPA) glutamate receptor (GluR) subunits is vital as they play key roles in the regulation of calcium permeability. We have examined the distribution of the calcium permeable AMPA glutamate receptor subunit GluR1 in the mouse visual cortex immunocytochemically. We compared this distribution to that of the calcium-binding proteins calbindin D28K, calretinin, and parvalbumin, and of GABA. The highest density of GluR1-immunoreactive (IR) neurons was found in layers II/III. Enucleation appeared to have no effect on the distribution of GluR1-IR neurons. The labeled neurons varied in morphology; the majority were round or oval and no pyramidal cells were labeled by the antibody. Two-color immunofluorescence revealed that 26.27%, 10.65%, and 40.31% of the GluR1-IR cells also contained, respectively, calbindin D28K, calretinin, and parvalbumin. 20.74% of the GluR1-IR neurons also expressed GABA. These results indicate that many neurons that express calcium-permeable GluR1 also express calcium binding proteins. They also demonstrate that one fifth of the GluR1-IR neurons in the mouse visual cortex are GABAergic interneurons.

• 한국어류학회지
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• 제23권1호
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• pp.1-9
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• 2011

ACP (annealing control primer)를 사용하여 DDRT (differential display reverse transcription)-PCR 방법으로 볼락의 성장단계에 따라 발현량 차이를 나타내는 DEG (differentially expressed gene)를 확보하였다. ACP 120개를 분석하여 18개월령 근육조직에서보다 6개월령 근육조직에서 발현량이 많은 DEG 16개와 6개월령 근육조직에서보다 18개월령 근육조직에서 발현량이 더 많은 DEG22개의 염기서열을 분석하였다. DEG 염기서열을 BLAST 검색한 결과, parvalbumin (PVALB) 등 18개의 유전자(PVALB, NDKB, TPM, TnI, GAPDH, CKM2, factor 2 SERF2, AMPD, TRICA, ARHGAP15, ESD, hsp70, COL1A2, GST, Midllip1, MYL1, SERCA1B, FTH1)와 69~95%의 상동성을 나타냈다. Real time PCR 분석법으로 6개월령 근육조직에서 발현량이 많은 DEG14와 PVALB 유전자의 성장단계별 발현양상을 조사한 결과, 볼락이 성장함에 따라 발현량이 감소하였으며, 특히 PVALB 유전자는 6개월령 이후에는 발현량이 극히 적었다. 6개월령 근육조직에서보다 18 개월령 근육조직에서 발현량에서 많았던 CKM2 유전자는 성장함에 따라 발현량이 계속 증가하였고, 4세 이후에는 발현량이 감소하였다. DEG의 조직특이적 발현양상을 분석한 결과, DEG14는 근육, 간, 신장, 및 비장조직에서 발현되었으며, PVALB 유전자는 근육과 신장조직에서 발현되었고, 간과 비장조직에서는 발현되지 않았다. CKM2 유전자는 근육, 신장 및 비장조직에서 발현되었고, 간 조직에서는 발현되지 않았다. PVALB 유전자의 mRNA 크기는 659 bp 이며, 110개의 아미노산으로 구성되어 있다. Parvalbumin과 CKM2 유전자는 성장속도가 빠른 어류 선발에 이용할 수 있는 분자마커 개발에 활용하고자한다.

• Environmental Analysis Health and Toxicology
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• 제24권4호
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• pp.321-332
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• 2009

본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

• BMB Reports
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• 제53권4호
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• pp.234-239
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• 2020

Epilepsy is a chronic neurological disease characterized by spontaneous recurrent seizures and caused by various factors and mechanisms. Malfunction of the olfactory bulb is frequently observed in patients with epilepsy. However, the morphological changes in the olfactory bulb during epilepsy-induced neuropathology have not been elucidated. Therefore, in the present study, we investigated the expression of parvalbumin (PV), one of the calcium-binding proteins, and morphological changes in the rat main olfactory bulb (MOB) following pilo-carpine-induced status epilepticus (SE). Pilocarpine-induced SE resulted in neuronal degeneration in the external plexiform layer (EPL) and glomerular layer (GL) of the MOB. PV immunoreactivity was observed in the neuronal somas and processes in the EPL and GL of the control group. However, six hours after pilocarpine administration, PV expression was remarkably decreased in the neuronal processes compared to the somas and the average number of PV-positive interneurons was significantly decreased. Three months after pilocarpine treatment, the number of PV-positive interneurons was also significantly decreased compared to the 6 hour group in both layers. In addition, the number of NeuN-positive neurons was also significantly decreased in the EPL and GL following pilocarpine treatment. In double immunofluorescence staining for PV and MAP2, the immunoreactivity for MAP2 around the PV-positive neurons was significantly decreased three months after pilocarpine treatment. Therefore, the present findings suggest that decreases in PV-positive GABAergic interneurons and dendritic density in the MOB induced impaired calcium buffering and reciprocal synaptic transmission. Thus, these alterations may be considered key factors aggravating olfactory function in patients with epilepsy.

• Clinical and Experimental Pediatrics
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• 제45권12호
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• pp.1551-1558
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• 2002

목 적 : 중증 측두엽간질환자와 kainic acid(KA)로 처리한 측두엽간질 동물모델에서 해마 치아이랑의 사이신경세포가 소실된다. 측두엽간질 마우스모델인 마우스해마의 기관형 배양에 의한 KA 간질모델에서 parvalbumin(PV)항체를 이용한 면역조직화학법으로 치아이랑에 분포하는 PV 면역반응성 사이신경세포를 감별염색하여 세포체 및 그 가지돌기의 형태학적 변화를 관찰함으로써 측두엽간질의 병태생리의 일단을 규명하고자 한다. 방 법 : 실험적 간질 모델은 C57/BL6 마우스의 해마박편을 이용한 기관형 배양에서 $10{\mu}M$ KA 투여로 유발시켰으며, PV 항체를 이용한 광학현미경적 면역조직화학법으로 치아이랑에 분포하는 PV 면역반응성 사이신경세포를 감별염색하여 형태학적 차이를 관찰하였고, 또한 이들 세포의 세포수를 계측하고 KA처리 후 배양하면서 시간경과(8, 24, 48, 72시간)에 따라 PV 면역반응성 사이신경세포의 형태학적 변화를 관찰하였다. 결 과: KA 처리를 하지 않고 배양된 해마조직박편(대조군)의 치아이랑에서 PV 면역반응성 세포는 가지돌기 나무가 잘 발달되어 있고 사이신경세포로서 이들은 주로 과립층과 이층 밑의 다형층에 산재하였다. $10{\mu}M$ KA에 1시간 정도 노출되었을 때 PV 면역반응성 사이신경세포의 돌기에는 염주상이 형성되었고 가지돌기는 가늘어져 있었다. PV 면역반응성 사이신경세포는 KA 처리 후 배양액에서 KA를 제거한 뒤 시간이 경과함에 따라 형태학적 회복을 보여주었다. KA 제거 후 8시간 회복군에서 세포돌기의 염주상은 볼 수 없었으며 가지돌기는 가늘어져 있었다. KA 제거 후 24, 48, 72시간 회복군에서도 염주상은 거의 볼 수가 없었으며 가지돌기의 두께도 회복되었다. PV 면역반응성 사이신경세포의 수는 대조군에 비하여 KA 처리군과 KA 제거 후 8시간 회복군에서는 통계학적으로 유의한 세포수의 감소가 있었으나 KA 제거 후 24시간 회복군, KA 제거 후 48시간 회복군 및 KA 제거 후 72시간 회복군에서는 대조군과 차이가 없 었다. 결 론: 이러한 결과는 KA에 의하여 유발된 치아이랑 사이신경세포의 세포소실이 일시적이며 가역적인 현상임을 말해주는 것이다.

• The Korean Journal of Physiology and Pharmacology
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• 제10권1호
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• pp.13-17
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• 2006

Experimentally induced cortical disorganization exhibits many anatomical features which are characteristic of cortical malformations in children with early-onset epilepsy. We used an immunocytochemical technique and extracellular field potential recordings from the dorsal hippocampus to determine whether the excitability of the CA1 pyramidal cells was enhanced in rats with exnerimentallv induced hippocampal dysplasia. Compared with control rats, the MAM-treated rats displayed a decrease of paired pulse inhibition. When $GABA_A$ receptor antagonists were blocked with $10{\mu}M$ bicuculline the amplitude of the second population spike of the MAM-treated of rats was similar to that of the first population spike, as was in the control rats. The MAM-treated rats had fewer somatostatin and parvalbumin-immunoreactive neurons than the control rats. These results suggest that the enhanced neuronal responsiveness of the in vivo recording of the CA1 in this animal model may involve a reduction of CA1 inhibition.

• 통합자연과학논문집
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• 제15권2호
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• pp.63-72
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• 2022

To elucidate the neurochemical characteristics of the midbrain periaqueductal gray (PAG), the distribution patterns of several neuroanatomical markers within the PAG were compared. Immunohistochemical staining for the intracellular calcium binding proteins including calbindin, calretinin, and parvalbumin and histochemical staining for cytochrome oxidase, acetylcholinesterase, and NADPH-diaphorase were performed in. Each chemical substance were localized in the specific subregions within PAG. Calbindin- immunoreactivity were selectively distributed in the dorsolateral PAG, the ventral half of lateral PAG, the ventralateral PAG, and supraoculomotor cap (Su3C) nucleus. Distribution of calretinin-immunoreactivity were generally similar with that of clabindin, but showed relatively low subregional selectivity. Parvalbumin-immunoreactivity was very poor within the PAG. High reactivity of cytochrome oxidase were found in the dorsomedial PAG and the lateral half of lateral PAG, in which calbindin- and calretinin-immunoreactive perikarya were scarcely observed. Acetylcholinesterase distribution was similar with that of cytochrome oxidase, and the difference was in the additional marking of of Su3C with acetylcholinesterase. Results of the present study provides data for the further subdivisions of the territory of the PAG compared to the presently accepted subregions within the PAG.

• 동의생리병리학회지
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• 제36권2호
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• pp.59-65
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• 2022

Rodent model for chronic cerebral hypoperfusion caused by bilateral carotid artery occlusion (BCAO) show clinically relevant evidences for vascular dementia and impairments of synaptic plasticity in the hippocampus. The purpose of this study was to evaluate effect of fermented garlic (F-Garlic) extract with NO metabolites on cognitive behaviors, synaptic plasticity, and molecular events in the hippocampus following BCAO. Adult male Sprague-Dawley rats were randomly divided three experimental groups into: control+water; BCAO+water; BCAO+F-Garlic. Animals were treated with oral administration of F-Garlic in tap water as a drinking water after surgery for 4 weeks. On passive avoidance test and Y-maze test, BCAO+water showed a significant decrease in step-through latency and spontaneous alteration, indicating deficit of hippocampal memory formation but the treatment of F-Garlic significantly increased these cognitive behaviors. In control+water, a robust increase in the amplitude of evoked field excitatory postsynaptic potentials was observed by theta burst stimulation to hippocampal neural circuit indicating formation of long-term potentiation (LTP) in the hippocampal CA1. BCAO+water showed a highly significant deficit in LTP induction 4 weeks after BCAO. On other hand, daily oral administration of F-Garlic extract caused the marked preservation of LTP induction. Moreover, parvalbumin was markedly reduced in the CA1, especially, in the stratum radiatum of BCAO+water. In contrast, BCAO+F-Garlic mitigate a significantly reduction of the parvalbumin. In summary, these results suggest that daily oral administration of F-Garlic extract can ameliorate cognitive memory deficit through the preservation of synaptic plasticity and interneurons integrity in the hippocampus in rodent model of chronic cerebral hypoperfusion.