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        저칼륨혈증 흰쥐 신장에서 Akt, p-Akt, ERK 및 p-ERK 단백발현의 변화

        배춘상(Choon Sang Bae),조혜정(Hye Jung Cho),안규윤(Kyu Yoon Ahn) 대한체질인류학회 2017 해부·생물인류학 (Anat Biol Anthropol) Vol.30 No.3

        저칼륨혈증은 신장의 형태학적 변화와 대사성 알칼리증을 유발시키는 것으로 알려져 있다. 이전 결과들은 저칼륨혈증의 병태생리학적 변화에 K+평형 조절 이온채널, pump 유전자, NF-E2-related factor 2 (Nrf2) 전사유전자를 포함한 다양한 유전자가 관여할 것이라는 가능성을 제시하였다. 이에 본 연구는 칼륨제한 식이 기간에 따른 흰쥐 신장 내 Akt와 p-Akt 및 ERK와 p-ERK의 발현 및 분포의 변화를 Western 분석과 면역조직화학 방법으로 관찰함으로써, 저칼륨혈증이 신장에서 AKT/ERK 인산화에 영향을 미치는지를 확인하였다. Western 분석소견에서 Akt 및 p-Akt 단백질 발현은 칼륨제한 식이가 진행될수록 증가하는 양상을 보였으며, ERK 및 p-ERK 단백질발현은 칼륨제한 식이 2주군에서 정상 식이군에 비해 약간 증가하는 양상을 보였다. 면역조직화학 소견에서 Akt 단백의 면역반응성은 먼쪽곱슬세관, 겉질곧은부분 및 속질곧은부분에서 중등도의 발현을 보였다. 칼륨제한 식이군의 Akt의 면역반응성은 칼륨제한 식이가 진행될수록 바깥속질집합관에서 현저히 증가하였다. 칼륨제한 식이군의 p-Akt의 면역반응성은 칼륨제한 식이 2주군의 먼쪽곱슬세관, 치밀반점과 속질곧은부분에서 증가하였고 토리쪽곱슬세관에서는 중등도의 발현을 보였다. 칼륨제한 식이군의 ERK의 면역반응성은 칼륨제한 식이 2주군과 3주군의 바깥속질집합관에서 현저히 증가하였으며 먼쪽곱슬세관, 겉질집합관에서는 증등도의 증가를 보였다. 칼륨제한 식이군의 p-ERK의 발현부위는 정상식이군과 비교해 차이가 없었으나 면역반응성은 칼륨제한 식이 2주군의 바깥속 질집합관의 핵에서 현저히 증가하였다. 이상의 결과 저칼륨혈증 시 p-Akt의 발현은 칼륨제한 식이가 길어질수록 점진적으로 증가하였지만, p-ERK의 발현은 칼륨제한 식이 2주군에서 현저히 증가되었다. 따라서 저칼륨 상태에서의 Akt 및 ERK 인산화의 촉진은 이온채널 및 이온수송체 유전자 조절뿐만 아니라 세포 내 신호전달에 있어 중요한 역할에 관여할 것임을 시사해 주었다. Hypokalemia causes metabolic alkalosis and morphological changes of the kidney. K+ balance is regulated not only by ion channels or pump gene, but also by various genes including NF-E2-related factor 2 (Nrf2). Previous study suggested the possibility that Akt and ERK kinase may be involved in Nrf2 transcriptional gene activation. In present study, we investigate the alterations of Akt, p-Akt, ERK, p-ERK protein in both normal kidney and K+-deficient diet kidney using Western blot analysis, and immunohistochemisrty. Our western blot data showed that the expression of Akt and p-Akt was increased gradually in K+-depleted diet (from 1W-3W) compared to normal group. The expression of ERK and p-ERK was markedly increased in K+-depleted diet 2W in comparison with normal group. Based on our immunostaining results, Akt protein immunoreactivity was prominently increased in outer medullary collecting duct, especially in K+-depleted diet 2 weeks. The localization of p-Akt proteins in K+-depleted groups was not different from normal group, but the immunoreactivity was significantly increased in distal convoluted tubule, macula densa and outer medullary thick ascending limb in K+-depleted diet 1 and 2 weeks groups. ERK protein immunoreactivity was prominently increased in outer medullary collecting duct, especially in K+-depleted diet 2 and 3 weeks. The localization of p-ERK proteins in K+-depleted groups was not different from normal group, but the immunoreactivity was prominently increased in the nucleus of outer medullary collecting duct especially in K+-depleted diet 2 weeks. Taken together, we suggest that the expression of p-Akt was gradually increased in K+-depleted groups of kidney, but the expression of p-ERK was markedly increased in K+-depleted diet 2 week group. Hence, the promotion of AKT and ERK phosphorylation in hypokalemic condition may be involved in the regulation of ion channels, ion transporters and subsequent intracellular signal transduction.

      • KCI등재
      • KCI등재

        고양이 송과체의 전자현미경적 연구

        최재권,배춘상,오창석,이정헌,Choi, Jae-Kwon,Bae, Choon-Sang,Oh, Chang-Seok,Lee, Jung-Hun 한국현미경학회 1992 Applied microscopy Vol.22 No.1

        Parenchyma of the cat pineal body consisted of pinealocytes and glial cells. The pinealocyte, predominant cell type, was characterized by having large mitochondria with pale matrix, abundant polyribosomes, moderately-developed Golgi apparatus, centrioles and occasional cilia. The pinealocyte had one thick and long cytoplasmic process at the one pole of the cell, and slender and shorter processes at the other pole, and in addition occasional short processes from the cell body. These processes contained longitudinally arranged microtubules, and a few mitochondria. Thick processes teminated as bulgings either in the intercellular process-rich area, or in the perivascular border which was formed by glial cell processes. These endings of pinealocyte processes had many small vesicles, mitochondria, and occasional dense bodies. Glial cells with abundant filaments of intermediate type and clear cytoplasmic matrix were fibrous astrocyte. Perikarya of the astrocytes had small and dense mitochondria, moderately developed Golgi apparatus, dense bodies and variable amount of intermediate filaments. Glial cell processes run through the intercellular spaces among the pinealocyte processes. Glial cell of protoplasmic type had no or a few filaments, but it had well-organized rough endoplasmic reticulum, dense mitochondria, well developed Golgi apparatus and many dense granules. Intercellular canaliculi formed by adjacent pinealocytes and glial cell processes were often noted. Within the parenchyma, sympathetic and parasympathetic axons and their endings were noted. These endings were present mostly in the intercellular spaces without having membrane specialization, however, in rare instances, ending with small clear and dense cored vesicles, and large dense cored vesicles formed specialized synapse with a pinealocyte process. Within the perivascular spaces nerve fibers and endings, Schwann cells and pericyte were noted. In rare case pinealocyte process penetrated into the perivascular space through the interuptions of glial border. These results suggest that pinealocyte of the cat has less significance in secretory function and is rather neural type of cell.

      • KCI등재
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      • KCI등재
      • KCI등재
      • KCI등재

        원숭이 외측슬상체배측핵에서 칼슘결합단백 Parvalbumin과 Calbindin-D 28K의 분포

        고승희,배춘상,박성식,Ko, Seung-Hee,Bae, Choon-Sang,Park, Sung-Sik 한국현미경학회 1994 Applied microscopy Vol.24 No.4

        The calcium-binding proteins (CaBP), parvalbumin (PV) and calbindin-D 28K (calbindin) are particularly abundant and specific in their distribution, and present in different subsets of neurons in many brain regions. Although their physiological roles in the neurons have not been elucidated, they are valuable markers of neuronal subpopulations for anatomical and developmental studies. This study is designed to characterize dorsal lateral geniculate nucleus (dLGN) neurons and axon terminals in terms of differential expression of immunoreactivity (IR) for two well-known CaBPs, PV and calbindin. The experiments were carried out on 6 adult monkeys. Monkeys were perfused under deep Nembutal anesthesia with 2% paraformaldehyde and 0.2% glutaraldehyde in 0.1M phosphate buffer. After removal, the brains were postfixed for 6-8 hr in 2% paraformaldehyde at $4^{\circ}C$ and infiltrated with 30% sucrose at $4^{\circ}C$. Thereafter, they were frozen in dry ice. Serial sections of the thalamus, at $20{\mu}m$, were made in the frontal plane with a sliding microtome. The sections were stained for PV and calbindin with indirect immunocytochemical methods. For electron microscopy, after infiltration with 30% sucrose the blocks of thalamus were serially sectioned at $50{\mu}m$ with a Vibratome in the coronal plane and stained immediately by indirect ABC methods without Triton X-100 in incubation medium. Stained sections were postfixed in 0.2% osmium tetroxide, dehydrated and flat-embedded in Spurr resin. The block was then trimmed to contain only a selected lamina or interlaminar space. The dLGN proper showed strong PV IR in fibers in all laminae and interlaminar zones. Particularly dense staining was noted in layers 1 and 2 that contain many stained fibers from optic tract. Neuronal cell body stained with PV was concentrated only in the laminae. In these laminae staining was moderate in cell bodies of all large and medium-sized neurons, and was strong in cell bodies of some small neurons together with their processes. Calbindin IR was marked in the neuronal cell body and neuropil in the S layers and interlaminar zones whereas moderate in the neuropil throughout the nucleus. Regional difference in distribution of PV and calbindin IR cell is distinct; the former is only in the laminae and the latter in both the S layer and interlaminar space. The CaBP-IR elements were confined to about $10{\mu}m$ in depth of Vibratome section. The IR product for CaBP was mainly associated with synaptic vesicle, pre- and post-synaptic membrane, and outer mitochondrial membrane and along microtubule. PV-IR was noted in various neuronal elements such as neuronal soma, dendrite, RLP, F, PSD and some myelinated or unmyelinated axons, and was not seen in the RSD and glial cells. Only a few neuronal components in dLGN was IR for calbindin and its reaction product was less dense than that of PV, and scattered throughout cytoplasm of soma of some relay neurons, and was also persent in some dendrite, myelinated axons and RLP. The RSD, F, PSD and glial elements were always non-IR for calbindin. Calbindin labelled RLP were presynaptic to unlabeled dendrite or dendritic spine and PSD. Calbindin-labeled dendrite of various sizes were always postsynaptic to unlabeled RSD, RLP or F. From this study it is suggested that dLGN cells of different functional systems and their differential projection to the visual cortex can be distinguished by differential expression of PV and calbindin.

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