• Title/Summary/Keyword: Mitochondria complex

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Screening Methods for Anti-senescence Activity in Dermal Fibroblasts under Pyruvate-deprivation Conditions

  • Kil, In Sup;Shim, Jinsup;Cho, Gayoung;Choi, Sowoong;Son, Eui Dong;Kim, Hyoung-June
    • Korea Journal of Cosmetic Science
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    • v.1 no.1
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    • pp.1-9
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    • 2019
  • The identification of compounds with anti-senescence activity in cell culture system is a first step in aging research. Given that pyruvate can be used energy source by conversion to acetyl-CoA in mitochondria, and protects cultured cell from various stress-induced cell damage and cell death, synthetic media (e.g., DMEM) often includes 1 mM pyruvate, which is very higher than the pyruvate concentration in human blood (approximately 30 ��M). However, the use of medium containing high concentration of pyruvate is not suitable for screening anti-senescence compounds, because pyruvate also protects against the cellular senescence of primary human dermal fibroblasts (NHDFs) through NAD+ generated during conversion to lactate. In this study, four extracts, i.e., Sprouted seed and fruit complex, Poncirus trifoliata fruit extract, Jaum balancing complex, and Prunus mume extract were used for evaluation of different anti-senescence effect in the absence or presence of 0.1 mM pyruvate, similar to the physiological pyruvate concentration. The senescence in NHDFs cultured with DMEM in the presence of 0.1 mM pyruvate (approximately the physiological concentration in human blood) is accelerated, as observed in pyruvate deprivation conditions. The cytotoxicity of the Poncirus trifoliata fruit extract was protected by pyruvate, and Jaum balancing complex and Prunus mume extract had anti-senescence activity in the presence of 0.1 mM pyruvate, but not in the absence of pyruvate. Given that pyruvate is a powerful protector against both cytotoxicity and cellular senescence, the screening of candidate agents for anti-senescence in high pyruvate conditions using an in vitro cell culture system is not valid. Therefore, we recommend the use of a low concentration of pyruvate to evaluate the anti-senescence effects of candidates, which is more similar to in vivo aging conditions than excessive stress-induced senescence models, to exclude the effect of excessive pyruvate in vitro.

Morphological Study on the Post-hatching Development of the Pineal Gland in Korean Pheasant (Phasianus colchicus Karpowi) (한국산 꿩 송과샘의 부화후 발달에 관한 형태학적 연구)

  • 이영훈;김인식;양홍현
    • Korean Journal of Poultry Science
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    • v.28 no.3
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    • pp.267-274
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    • 2001
  • The pineal gland of the bird occupies a key position in the phylogenetic evolution of this organ. Therefore, the purpose of this study was to investigate the developmental changes of the pineal gland during post-hatching period in Korean pheasant. The pheasants were sacrificed at 1-day-, 1-month-, 2-month-, and 6-month-old after hatching. The morphological characteristics of a pineal glands were determined in all pheasants using light microscope, and transmission electron microscope. Connective tissue originated from the capsule divided the pineal parenchyma into incomplete lobules. The parenchyma was consisted of pinealocytes and supportive cells. These parenchymal cells were arranged in the forms of solid lobules as well as incomplete follicles. At the follicular lumen, membraneous lamellar complexes and blob -like structures were present. Pinealocyte, a predominent cell type, had euchromatic nucleus, and showed the segmental organization. The bulbous apical portion had scanty free ribosomes and occasional cilia associated with basal bodies. The constricted neck, transitional portion from apical to pericarya had junctional complexes with adjacent supportive cells, and had microtubules. Cell body contained abundant mitochondria, well-developed Golgi complex, rough endoplasmic reticulum (RER) and free ribosomes. Basal processes extended from the base of the cell soma toward the basal lamina and contained 60∼90 nm dense cored vesicles. Supportive cells, another major type of the parenchyma, were characterized by the dense and elongated nucleus, and contained moderate number of mitochondria, RER, developed Golgi complex, free ribosomes and a few dense bodies in the perinuclear cytoplasm. Slender processes of supportive cells interposed between the pinealocytes and often bordered the basal region of the parenchyma. These results indicate that the pinealocytes of the pheasant are not rudimentary photoreceptor cells, and appear to have secretory function. Further studies will be required to confirm the morphological characteristics of pineal gland in adult pheasant during breeding and nonbreeding season.

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Comparative Ultrastructure of the Acinar Cell and Secretory Granules of Parotid Salivary Gland in the Lesser White-toothed Shrew, Crocidura suaveolens and the Big White-toothed Shrew, C. lasiura (작은땃쥐 Crocidura suaveolens와 우수리땃쥐 C. lasiura의 이하선 선세포와 분비과립의 미세구조 비교)

  • Jeong, Soon-Jeong;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.35 no.4
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    • pp.91-97
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    • 2005
  • The acinar cells and secretory granules of the parotid salivary gland were examined in the lesser white toothed shrew, Crocidura suaveolens and the big white-toothed shrew, C. lasiura. The parotid gland of both species were a serous gland having only one kind of serous acinar cells, and had conventional arrangement of acini and intercalated, granular and striated ducts. In case of C. suaveolens, serous acinar cells had well developed rER, prominent Golgi complex, several large mitochondria and abundant moderate dense secretory granules with various stages of the maturing or fusing process. Immature acinar secretory granules were only or mainly filled with fine strong dense specks and had an indistinct limiting membrane, and mature granules were filled with homogeneous pale large round center and had fine strong dense specks at the periphery of the homogeneous pale center and a distinct limiting membrane. In case of C. lasiula, serous acinar cells had well developed rER, prominent Golgi complex, several large mitochondria and abundant dense secretory granules with maturing or fusing process. Immature acinar secretory granules were only filled with pale rough specks and had an indistinct limiting membrane, and mature granules were only filled with rough dense specks and had a distinct limiting membrane. Eventually The acinar secretory granules of C. suaveolens were seen moderate at the light and ultrastuctural level, those of C. lasiura were strong dense at the light microscopic level and dense at the ultrastructural level.

Ultrastructure and Dehydrogenase Activity on the Differentiation of the Cerebral Nerve Cell in the Chick Embryo (II) (계배 대뇌의 신경세포 분화에 따른 탈수소효소 활성 및 미세구조(II))

  • Kim, Saeng-Gon
    • Applied Microscopy
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    • v.29 no.4
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    • pp.459-470
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    • 1999
  • To investigate the changes during the differentiation of the cerebral neurons of the embryogenic day (ED) 9 and 10, investigated the ultrastructural changes in the cerebral neurons by Electromicroscope, also cerebral protein, the activity of dehydronases (LDH, MDH and SDH) and changes of adenosine triphosphate concentration were analyzed, the result obtained are as follows. In the ultrastructural changes in the cerebral neurons, chromatin in 9 day-old chick embryos are comparatively distributed to even in neucleoplasm and could investigate very prominently that nuclear membrane is double-layer. Esperially, Rough endoplasmic reticulum (RER) and Golgi complex are developed well, also polysome is investigated and synaptic vesicles were scattered. In 10 day-old chick embryos, chromatin evenly spread and nuclear membrane could be differentiated prominently. Rough endoplasmic reticulum (RER) contain cytoplasm, mitochondria and Golgi complex are comparatively developed well. In 9 day-old cultural group of chick embryo cerebrum were separated 37 polypeptide bands and In 10 day-old cultural group of chick embryo cerebrum were separated 38 poly -peptide bands. The more culture time increase, the more the activity of dehydronases (LDH, MDH and SDH) increase. LDH activity was 11.07 (9th day) and 12.12 (10th day), MDH activity was 11.89 (9th day) and 13.44 (10th day) and SDH activity was 8.45 (9th day) and 10.52 (10th day) respectively. The ATP concentration degreesed 10 day-old cultural group than 9 day-old cultural group.

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Effect of Testoeterone and Cyclic AMP-theophylline on Ultrastructure and Several Enzymes' Activities in Rat Epididymis (Testosterone과 Cyclic AMP-theophyllin이 흰쥐 부정소의 미세구조 및 몇 가지 효소활성에 미치는 영향)

  • 정경순;최임순
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.35-48
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    • 1988
  • This study was made to investigate the effect of testosterone and cyclic-AMP (cAMP) on rat epididymis. Peritoneai injections of testosterone and cAMP to rats were earned out The activities of acid phosphatase (ACP), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were measured and ultrastructural changes of luminal epithelial cells were observed. As a result, the activity of ACP was significanily decreased on the third day, that of ALP on the fifth day and that of LDH on the seventh day respectively in castrated group. In addition, the activities of ACP and ALP were significanily increased when treated with testosterone for 5 days, that of LDH when treated with testosterone for 7 days. In case of cAMP and cAMP - theophylline injection, the activities of ACP and LDH were increased but the range of increase was of no significance. However a significant increase in the activity of ALP was seen on both cases. On electron microscopic examination, gradually deformed Golgi complex, destructed mitochondria and disrupted stereociha were observed in castrated group. In case of testosterone injection, disrupted Golgi complex, mitochondria and stereocilia showed recovery. When cAMP and cAMP-theophylline were injected as an alternative, various cytoplasmic organelles as well as Golgi com- plex were recovered but stereocilia remained unrecovered.

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Spermatogenesis and Sperm Ultrastructure of the Equilateral Venus, Gomphina veneriformis (Bivalvia: Veneridae) (대복, Gomphina veneriformis의 정자형성과정 및 정자 미세구조)

  • Park, Chae-Kyu;Park, Jung-Jun;Lee, Jeong-Yong;Lee, Jung-Sick
    • Applied Microscopy
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    • v.32 no.4
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    • pp.303-310
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    • 2002
  • Spermatogenesis and sperm ultrastructure are investigated by means of light and transmission electron microscopy in the equilateral venus, Gomphina veneriformis which is dominant bivalve in the east coast of Korea. In the active spermatogenic season, testis consists of numerous spermatogenic follicles which is contains germ cells in the different developmental stage. The spermatogonia attached to spermatogenic follicle wall and has a large nucleus with electron-dense nucleolus. The spermatocytes are characterized by appearance of synaptonemal complex and well-developed Golgi complex. Nucleus of spermatid consists of numerous heterogeneous granules with high electron density. Karyoplasmic condensation, acrosome and flagellum formations are observed during spermiogenesis. Testicular matured sperms of sperm bundle consists of head, midpiece and tail. The head is about $8.5{\mu}m$ long and comprises a long nucleus and a bullet-like acrosome ($8.5{\mu}m$ in length). Acrosomal rod of microfilaments is observed in the lumen between nucleus and acrosome. The midpiece has four mitochondria. And tail has the typical '9+2' microtubule system.

Ultrastructural Study on the Development of Male Germ Cell of the Olive Flounder, Paralichthys olivaceus (Teleostei: Pleuronectidae) (넙치 (Paralichthys olivaceus)의 웅성생식세포 발달에 관한 미세구조적 연구)

  • Kim, Jae-Won;Kim, Bong-Seok;Choi, Cheol-Young;Lee, Jung-Sick
    • Applied Microscopy
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    • v.33 no.3
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    • pp.243-250
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    • 2003
  • Ultrastructural changes of the male germ cells and structure of spermatozoa in Paralichthys olivaceus were examined by means of the light and transmission electron microscopes. The spermatogonium has a large nucleus with a single nucleus with a single nucleolus in the interphase. Primary spermatocytes are identified by the formation of the synaptonemal complex in the karyoplasm. The secondary spermatocytes are more concentrated and contains numerous cell organelle in the cytoplasm. The nucleus of spermatid in spermiogenesis is more condensed in the karyoplasm, and show spherical structure in shape. Mitochondria of the spermatids are observed in the lower portion of the nucleus. The spermatozoon consists of the head, mid piece and tail. The acrosome is not observed in the head. Axial filaments of the flagellum consists of nine pairs of the peripheral microtubules and one pair of the central microtubules.

An Electron Microscopic Study on the Cell Wall Regeneration of Culture Panax ginseng Callus Protoplast (인삼 캘러스 원형질체의 배양에 따른 세포벽 재생의 전자현미경적 연구)

  • 박종범
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.495-500
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    • 1998
  • Ultrastructural changes of the isolated and cultured protoplasts from ginseng (Panax ginseng C. A. Meyer) callus were studies with electron microscopy. In the 3-day-cultured protoplasts, the cell organelles such as rough endoplasmic reticulum, ribosome, Golgi complex, mitochondria, proplastid increased in number and observed microtubules. Many vesicles derived from the Golgi complex were evenly distributed in the cytoplasm. Some of such vesicles protruded the outer surface of the plasmalemma, and formed the protuberances. Vacuole derived from endoplasmic reticulum included Golgi vesicles by the invagination of vacuoles. These vacuoles migrated toward the plasmalemma by a fusion process (exocytosis), after fusing the plasmalemma the cell wall materials released from the outer surface of the plasmalemma, and lastly deposited on the plasmalemma. Proplastids containing many starch grains, and microtubules parallel to the plasmalemma were observed near the plasmalemma. Connected fibrils which were observed on the outer surface of the 3-day-cultured protoplast were interpreted as the component of cellulose.

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Risk factors of ocular involvement in children with mitochondrial respiratory chain complex defect

  • Chae, Jung-Hyun;Lee, Jung-Hun;Kim, Kyo-Ryung;Byeon, Suk-Ho;Lee, Young-Mock;Kang, Hoon-Chul;Lee, Joon-Soo;Kim, Heung-Dong
    • Clinical and Experimental Pediatrics
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    • v.53 no.12
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    • pp.994-999
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    • 2010
  • Purpose: Mitochondrial dysfunction can present with various symptoms depending on the organ it has affected. This research tried to analyze the ophthalmologic symptoms and ophthalmologic examination (OE) results in patients with mitochondrial disease (MD). Methods: Seventy-four patients diagnosed with mitochondrial respiratory chain complex defect with biochemical enzyme assay were included in the study. They were divided into 2 groups based on the OE results by funduscopy and were analyzed on the basis of their clinical features, biochemical test results, morphological analysis, and neuroimaging findings. Results: Thirty-seven (50%) of the 74 MD patients developed ophthalmologic symptoms. Abnormal findings were observed in 36 (48.6%) patients during an OE, and 16 (21.6%) of them had no ocular symptoms. Significantly higher rates of prematurity, clinical history of epilepsy or frequent apnea events, abnormal light microscopic findings in muscle pathology, diffuse cerebral atrophy in magnetic resonance imaging, and brainstem hyperintensity and lactate peaks in magnetic resonance spectroscopy were noted in the group with abnormal OE results. Conclusion: Although the ophthalmologic symptoms are not very remarkable in MD patients, an OE is required. When the risk factors mentioned above are observed, a more active approach should be taken in the OE because a higher frequency of ocular involvement can be expected.

Neurotoxin-Induced Pathway Perturbation in Human Neuroblastoma SH-EP Cells

  • Do, Jin Hwan
    • Molecules and Cells
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    • v.37 no.9
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    • pp.672-684
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    • 2014
  • The exact causes of cell death in Parkinson's disease (PD) remain unknown despite extensive studies on PD.The identification of signaling and metabolic pathways involved in PD might provide insight into the molecular mechanisms underlying PD. The neurotoxin 1-methyl-4-phenylpyridinium ($MPP^+$) induces cellular changes characteristic of PD, and $MPP^+$-based models have been extensively used for PD studies. In this study, pathways that were significantly perturbed in $MPP^+$-treated human neuroblastoma SH-EP cells were identified from genome-wide gene expression data for five time points (1.5, 3, 9, 12, and 24 h) after treatment. The mitogen-activated protein kinase (MAPK) signaling pathway and endoplasmic reticulum (ER) protein processing pathway showed significant perturbation at all time points. Perturbation of each of these pathways resulted in the common outcome of upregulation of DNA-damage-inducible transcript 3 (DDIT3). Genes involved in ER protein processing pathway included ubiquitin ligase complex genes and ER-associated degradation (ERAD)-related genes. Additionally, overexpression of DDIT3 might induce oxidative stress via glutathione depletion as a result of overexpression of CHAC1. This study suggests that upregulation of DDIT3 caused by perturbation of the MAPK signaling pathway and ER protein processing pathway might play a key role in $MPP^+$-induced neuronal cell death. Moreover, the toxicity signal of $MPP^+$ resulting from mitochondrial dysfunction through inhibition of complex I of the electron transport chain might feed back to the mitochondria via ER stress. This positive feedback could contribute to amplification of the death signal induced by $MPP^+$.