• Molecules and Cells
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• 제19권3호
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• pp.382-390
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• 2005

Calcium-binding proteins are thought to play important roles in calcium buffering. The present study investigated the effects of ischemia and reperfusion on calbindin D28K, calretinin, and parvalbumin immunoreactivity in the ganglion cell layer of the rabbit. Rabbits were administered ischemic damage by increasing the intraocular pressure. After 60 and 90 min of ischemia, reperfusion (7 d) was allowed to occur. The b-wave of the electroretinogram (ERG) was reduced by more than 50% and almost 80% in retina given ischemia for 60 and 90 min, respectively. The oscillatory potential (OPs) wave was reduced approximately 50% at 60 min ischemia and 70% at 90 min ischemia. In both normal and ischemic-treated retina, calcium-binding protein immunoreactivity was seen in many cells in the ganglion cell layer. In eyes subjected to 60 min ischemia, there was a decrease of the density of calbindin D28K- (8.29%), calretinin- (14.44%), and parvalbumin- (26.83%) immunoreactive (IR) cells compared to the control retina. In eyes subjected to 90 min ischemia, there was a higher decrease of the density of calbindin D28K- (18.48%), calretinin- (33.59%), and parvalbumin- (54.26%) IR cells than at 60 min. Some calcium-binding protein-IR neurons, especially calretinin-IR neurons, showed aggregations that were abnormally packed together in retina subjected to ischemia for 90 min. The results show that calbindin D28K-, calretinin-, and parvalbumin-IR cells in the ganglion cell layer are susceptible to ischemic damage and reperfusion. The degree of reduction varied among different calcium-binding proteins and ischemic damage times. These results suggest that calbindin D28K-containing neurons are less susceptible to ischemic damage than calretinin- and parvalbumin-containing neurons in the ganglion cell layer of rabbit retina.

• 대한수의학회지
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• 제39권1호
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• pp.34-44
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• 1999

Ischemic damage in the selectively vulnerable populations of neurons is thought to be caused by an abnormal accumulation of intracellular calcium. It has been reported that the neurons, expressing specific calcium binding proteins, might effectively control intracellular calcium concentrations because of a high capacity to buffer intracellular calcium in the brain ischemic condition. It is uncertain that parvalbumin, one of the calcium binding proteins, can protect the neurons from the cerebral ischemic damage. Recently, treatment of kappa opioid agonists increased survival rate, improved neurological function, and decreased tissue damage under the cerebral ischemic condition. Many evidences indicate that these therapeutic effects might result from regulation of calcium concentration. This study was designed to analyze the changes of number in parvalbumin-positive neurons after cerebral ischemic damage according to timepoints after cerebral ischemic induction. In addition, we evaluated the effect of GR89696 (kappa opioid agonist) or naltrexone(non selective opioid antagonist) on the changes of number in parvalbumin expressing neurons under ischemic condition. Cerebral ischemia was induced by occluding the common carotid artery of experimental animals. The hippocampal areas were morphometrically analyzed at different time point after ischemic induction(1, 3, 5 days) by using immuno-histochemical technique and imaging analysis system. The number of parvalbumin-positive neurons in hippocampus was significantly reduced at 1 day after ischemia(p<0.05). Furthermore, the number of parvalbumin-immunoreactive neurons was dramatically reduced at 3 and 5 days after cerebral ischemic induction(p<0.05) as compared to 1 day group after ischemia, as well as sham control group. Significant reduction of parvalbumin positive neurons in CA1 region of hippocampus was observed at 1 day after cerebral ischemic induction. However, significant loss of MAP2 immunoreactivity was observed at 3 day after cerebral ischemia. The loss of parvalbumin-positive neurons and MAP2 immunoreactivity in CA1 region was prevented by pre-administration of GR89696 compared to that of saline-treated ischemic group. Furthermore, protective effect of GR89696 partially reversed by pre-treatment of naltrexone. These data indicate that parvalbumin-positive neurons more sensitively responded to cerebral ischemic damage than MAP2 protein. Moreover, this loss of parvalbumin-positive neurons was effectively prevented by the pretreatment of kappa opioid agonist. It was also suggested that the changes of number in parvalbumin-positive neurons could be used as the specific marker to analyze the degree of ischemic neuronal damage.

• 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.

• 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.

• 대한임상독성학회지
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• 제19권2호
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• pp.100-109
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• 2021

Purpose: The purpose of this study was to investigate effect of N-acetylcysteine (NAC) on the injury of putative parvalbumin positive interneurons defined by molecular marker and hippocampal long-term potentiation (LTP), a marker of neural plasticity following acute carbon monoxide (CO) poisoning. Methods: Adult Sprague-Dawley rats were exposed to 1100 ppm CO for 40 minutes followed by 3000 ppm CO for 20 minutes. Animals received daily intraperitoneal injection of NAC (150 mg/kg) for 5 days after CO exposure. Changes in learning and spatial memory were evaluated by Y-maze test 5 days after the poisoning. In vivo LTP in hippocampal CA1 area was evaluated by using extracellular electrophysiological technique. Immunohistochemical staining were adopted to observe expressional damages of parvalbumin (PV) immunoreactive interneurons in the hippocampus following the poisoning. Results: Acute CO intoxication resulted in no changes in memory performance at Y-maze test but a significant reduction of LTP in the in hippocampal CA1 area. There was also a significant reduction of PV (+) interneurons in the hippocampal CA1 area 5 days after CO poisoning. Daily treatment of NAC significantly improved hippocampal LTP impairment and reduced immunoreactivity for PV in the hippocampus following the acute CO poisoning. Conclusion: The results of this study suggest that reduction of hippocampal LTP and PV (+) interneurons in the hippocampus is sensitive indicator for brain injury and daily NAC injections can be the alternative therapeutics for the injury induced by acute CO poisoning.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1068-1074
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• 2007

파브알부민(pa π albumin)은 망막의 다양한 세포타입에서 분포하고 있다. 본 연구팀은 이전연구에서 박쥐 망막의 내핵층에서의 파브알부민의 분포를 보고하였다. 현재 연구에서 본 연구팀은 한국관박쥐 (Rhinolophus ferrumequinum) 망막의 신경절세포층에 존재하는 파브알부민을 함유하는 신경세포를 규명하였고, 이들 세포의 분포양상을 조사하였다. 실험 결과,파브알부민의 면역반응성은 신경절세포층의 다수 세포에서 발견되었으며, 이들 세포는 주로 중간형 이상 크기의 세포체를 가지고 있었다. 조사된 세포체의 직경은 12.35 - 19.12 ${\mu}m$ 의 범위를 가지며 (n=166), 신경섬유층의 섬유 역시 염색되는 것으로 보아, 파브알부민을 함유하는 신경절세포는 대부분이 중간형이상 크기의 신경절세포임을 뒷받침하고 있다. NND (nearest neighbor distance) 분석을 통해서 본, 평균 NND는 59.57 에서 62.45 ${\mu}m$ 로 나타났으며, 평균 RI (regularity index) 는 2.95 ${\pm}$ 0.3 (mean${\pm}$s.d., n=4) 으로 계산되었다. 이를 종합해보면, 파브알부민은 한국관박쥐 망막의 신경절세포층에서 중간형이상 크기의 신경절세포에서 주로 발현하고 있으며, 이들은 규칙적인 배열을 가진 채 잘 조직화된 분포양상을 보여주고 있음을 알 수 있었다. 이러한 결과들은, 아직까지 명확하게 규명되어 있지 못한 박쥐의 시각에 대한 이해에 중요하게 적용될 수 있을 것이라고 사료된다.

• 한국안광학회지
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• 제19권2호
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• pp.271-277
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• 2014

목적: 본 연구의 목적은 브라질산 주머니쥐(Monodelphis domestica) 망막에서의 calretinin 면역반응성을 연구하는데에 있다. 칼슘결합단백질 calretinin은 칼슘매개-신호전달에서 중요한 역할을 한다고 알려져 있다. 방법: 표준 면역조직화학법을 이용하여 브라질산 주머니쥐 망막을 대상으로 실험하였다. 결과: 브라질산 주머니쥐 망막에서 calretinin은 일부의 수평세포, AII 무축삭세포, 일부의 신경절세포에서 면역반응성을 보였다. 특히, calretinin 면역반응성을 가지는 모든 AII 무축삭세포는 또 다른 칼슘결합단백질인 parvalbumin을 발현하였다. 결론: 다른 포유류 망막과 다를 바 없이, 브라질산 주머니쥐 망막에서도 AII 무축삭세포에서 calretinin 면역반응성이 관찰되었다. 이를 통해 calretinin이 브라질산 주머니쥐 망막에서도 AII 무축삭세포의 marker로 이용될 수 있다는 점을 확인하였다.

• 대한수의학회지
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• 제41권2호
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• pp.139-146
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• 2001

This study was carry out to identify the distribution of calcium binding proteins; calbindin(CB), calretinin(CR) and parvalbumin(PA) in the suprachiasmatic nucleus(SCN) of the Korean native goat by immunohistochemical methods. The expression of substance P(SP), calcitonin gene-related peptide(CG-RP), neuropeptide Y(NPY), vasoactive intestinal polypeptide(VIP) and galanin(GAL) were also investigated. CR-immunoreactivity was found in both of the cell bodies and fibers in the SCN, which the CB-immunoreactivity was observed only in the fibers. The immunoreactivity for VIP was observed in both the cell bodies and fibers, but SP-, NPY, GAL-immunoreactivities were only found in the fibers. CGRP-immunoreactivity was not seen in cell body and fibers. These results suggest that VIP, SP, NPY and GAL play a neuromodulatory or/ and neurotransmitter roles in cooperation with CB and CR in SCN of the Korean native goat.

• The Korean Journal of Physiology and Pharmacology
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• 제27권1호
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• pp.113-125
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• 2023

It has been reported that stressful events in early life influence behavior in adulthood and are associated with different psychiatric disorders, such as major depression, post-traumatic stress disorder, bipolar disorder, and anxiety disorder. Maternal separation (MS) is a representative animal model for reproducing childhood stress. It is used as an animal model for depression, and has well-known effects, such as increasing anxiety behavior and causing abnormalities in the hypothalamic-pituitary-adrenal (HPA) axis. This study investigated the effect of MS on anxiety or aggression-like behavior and the number of GABAergic neurons in the hippocampus. Mice were separated from their dams for four hours per day for 19 d from postnatal day two. Elevated plus maze (EPM) test, resident-intruder (RI) test, and counted glutamic acid decarboxylase 67 (GAD67) or parvalbumin (PV) positive cells in the hippocampus were executed using immunohistochemistry. The maternal segregation group exhibited increased anxiety and aggression in the EPM test and the RI test. GAD67-positive neurons were increased in the hippocampal regions we observed: dentate gyrus (DG), CA3, CA1, subiculum, presubiculum, and parasubiculum. PV-positive neurons were increased in the DG, CA3, presubiculum, and parasubiculum. Consistent with behavioral changes, corticosterone was increased in the MS group, suggesting that the behavioral changes induced by MS were expressed through the effect on the HPA axis. Altogether, MS alters anxiety and aggression levels, possibly through alteration of cytoarchitecture and output of the ventral hippocampus that induces the dysfunction of the HPA axis.

• 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.