• Title/Summary/Keyword: Subcellular Characteristics

Search Result 19, Processing Time 0.027 seconds

Subcellular Characterization of Porcine Oocytes with Different Glucose-6-phosphate Dehydrogenase Activities

  • Fu, Bo;Ren, Liang;Liu, Di;Ma, Jian-Zhang;An, Tie-Zhu;Yang, Xiu-Qin;Ma, Hong;Zhang, Dong-Jie;Guo, Zhen-Hua;Guo, Yun-Yun;Zhu, Meng;Bai, Jing
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.28 no.12
    • /
    • pp.1703-1712
    • /
    • 2015
  • The in vitro maturation (IVM) efficiency of porcine embryos is still low because of poor oocyte quality. Although brilliant cresyl blue positive (BCB+) oocytes with low glucose-6-phosphate dehydrogenase (G6PDH) activity have shown superior quality than BCB negative (-) oocytes with high G6PDH activity, the use of a BCB staining test before IVM is still controversial. This study aimed to shed more light on the subcellular characteristics of porcine oocytes after selection using BCB staining. We assessed germinal vesicle chromatin configuration, cortical granule (CG) migration, mitochondrial distribution, the levels of acetylated lysine 9 of histone H3 (AcH3K9) and nuclear apoptosis features to investigate the correlation between G6PDH activity and these developmentally related features. A pattern of chromatin surrounding the nucleoli was seen in 53.0% of BCB+ oocytes and 77.6% of BCB+ oocytes showed peripherally distributed CGs. After IVM, 48.7% of BCB+ oocytes had a diffused mitochondrial distribution pattern. However, there were no significant differences in the levels of AcH3K9 in the nuclei of blastocysts derived from BCB+ and BCB- oocytes; at the same time, we observed a similar incidence of apoptosis in the BCB+ and control groups. Although this study indicated that G6PDH activity in porcine oocytes was correlated with several subcellular characteristics such as germinal vesicle chromatin configuration, CG migration and mitochondrial distribution, other features such as AcH3K9 level and nuclear apoptotic features were not associated with G6PDH activity and did not validate the BCB staining test. In using this test for selecting porcine oocytes, subcellular characteristics such as the AcH3K9 level and apoptotic nuclear features should also be considered. Adding histone deacetylase inhibitors or apoptosis inhibitors into the culture medium used might improve the efficiency of IVM of BCB+ oocytes.

Subcellular Distribution and Characteristics of Polyphenol Oxidase from Perilla frutescens Leaves (들깨잎 polyphenol oxidase의 세포내 분포 및 특성)

  • 김안근;김유경
    • YAKHAK HOEJI
    • /
    • v.43 no.6
    • /
    • pp.709-715
    • /
    • 1999
  • Polyphenol oxidase (PPO) activity in 200×g (cell wall), 4,000×g (plastid), 100,000×g (mitochondrial) and soluble fractions of the perilla leaves was monitored in the upper, middle and lower sections of the plant. In the course of plant growth, PPO activities in plastid and mitochondrial fractions were decreased, while those in cell wall fraction were maintained. During growing process, specific activities and PPO activities of each fraction were decreased, while total phenol content were decreased in middle (middle) and then increased in later stage (lower). Cell wall, plastid, mitochondrial (pellet) and soluble fraction had slightly different pH optima and substrate specificities. Isoenzyme patterns were identical in two bands for PPO activity in different subcellular fractions. Their molecular weights were 37KD and 48KD respectively.

  • PDF

Functional Characteristics of Cytoplasmic and Periplasmic Photobacterium leiognathi CuZnSOD (PSOD) in Escherichia coli SOD Double Mutants (Escherichia coli SOD 이중 돌연변이체내에서 세포질과 Periplasm에 분포하는 Photobacterium leiognathi CuZnSOD(PSOD)의 기능적 특성)

  • Kim, Young-Gon;Yang, Mi-Kyung
    • Korean Journal of Microbiology
    • /
    • v.32 no.2
    • /
    • pp.139-146
    • /
    • 1994
  • Protective effects on subcellular localization of Photobacterium leiognathi CuZnSOD(PSOD) were examined in Escherichia coli SOD mutant cells on the treatment of paraquat, heat shock $(37^{\circ}C{\to}42^{\circ}C{\to})$, hydrogen peroxide and copper sulfatem respectively. The physiological characteristics of the periplasmic and cytoplasmic PSOD localized differently are dependent on the conditions in this experiment. Cells expressing SOD periplasmically in the treatments of paraquat and $H_2O_2$ respectively were somewhat better protective effects cells expressiong SOD cytoplasmically at comparable level and SOD expression level showed, the most consistently important variable. However, this was reversed in the treatments of heat shock and $CuSO_4$, respectively.

  • PDF

Sequence analysis of LSU rDNA of Alexandrium tamarense/catenella complex from Korean coastal waters

  • Kim, Keunyong;Kim, Chang-Hoon
    • Proceedings of the Korean Society of Fisheries Technology Conference
    • /
    • 2001.05a
    • /
    • pp.252-254
    • /
    • 2001
  • A great deal of effort has been put into the identification of Alexandrium tamarense/fundyense/catenella complex by understanding correlation between morphological and subcellular characteristics. To date, the most promising tool for the study of these species is sequence analyses of rRNA genes that have been useful for various organisms' taxonomy and phylogeny, and its application such as in situ hybridization. (omitted)

  • PDF

Production and identification of antisera against mu-opioid receptor using synthetic peptide epitope (Synthetic peptide를 이용한 mu-opioid receptor에 대한 항혈청의 생산과 검정)

  • Lee, Jang-hern;Kwon, Young-bae;Han, Ho-jae
    • Korean Journal of Veterinary Research
    • /
    • v.39 no.1
    • /
    • pp.45-54
    • /
    • 1999
  • In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398 of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide synthesizer. The specificity and identification of the antisera were tested by analysis of transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR. MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as endoplasmic reticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity was abolished when anti-MOR sera were preincubated with the peptide against which they were raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which was determined to be within the last amino acid, 391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was observed in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain areas showed the mirrored pattern observed in auto radiographic studies of mu-opioid binding as well as a pattern similar to that seen by is situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future.

  • PDF

Nano-Resolution Connectomics Using Large-Volume Electron Microscopy

  • Kim, Gyu Hyun;Gim, Ja Won;Lee, Kea Joo
    • Applied Microscopy
    • /
    • v.46 no.4
    • /
    • pp.171-175
    • /
    • 2016
  • A distinctive neuronal network in the brain is believed to make us unique individuals. Electron microscopy is a valuable tool for examining ultrastructural characteristics of neurons, synapses, and subcellular organelles. A recent technological breakthrough in volume electron microscopy allows large-scale circuit reconstruction of the nervous system with unprecedented detail. Serial-section electron microscopy-previously the domain of specialists-became automated with the advent of innovative systems such as the focused ion beam and serial block-face scanning electron microscopes and the automated tape-collecting ultramicrotome. Further advances in microscopic design and instrumentation are also available, which allow the reconstruction of unprecedentedly large volumes of brain tissue at high speed. The recent introduction of correlative light and electron microscopy will help to identify specific neural circuits associated with behavioral characteristics and revolutionize our understanding of how the brain works.

Computational approaches for molecular characterization and structure-based functional elucidation of a hypothetical protein from Mycobacterium tuberculosis

  • Abu Saim Mohammad, Saikat
    • Genomics & Informatics
    • /
    • v.21 no.2
    • /
    • pp.25.1-25.12
    • /
    • 2023
  • Adaptation of infections and hosts has resulted in several metabolic mechanisms adopted by intracellular pathogens to combat the defense responses and the lack of fuel during infection. Human tuberculosis caused by Mycobacterium tuberculosis (MTB) is the world's first cause of mortality tied to a single disease. This study aims to characterize and anticipate potential antigen characteristics for promising vaccine candidates for the hypothetical protein of MTB through computational strategies. The protein is associated with the catalyzation of dithiol oxidation and/or disulfide reduction because of the protein's anticipated disulfide oxidoreductase properties. This investigation analyzed the protein's physicochemical characteristics, protein-protein interactions, subcellular locations, anticipated active sites, secondary and tertiary structures, allergenicity, antigenicity, and toxicity properties. The protein has significant active amino acid residues with no allergenicity, elevated antigenicity, and no toxicity.

Immunologic Mechanism of Experimental and Therapeutic Ultraviolet B Responses

  • Lew, Wook
    • IMMUNE NETWORK
    • /
    • v.2 no.2
    • /
    • pp.65-71
    • /
    • 2002
  • The immunological mechanism of the responses to ultraviolet (UV) B radiation in mouse models were investigated by the suppression of contact hypersensitivity (CHS) and delayed type hypersensitivity (DTH), and susceptibility to infection. However, there are some differences in immune suppression according to the different models as well as the irradiation protocols. Therefore, this review focused on the differences in the suppressive effects on CHS and DTH, and susceptibility to infection in relation to the different in vivo models. Recent advances in cytokine knockout mice experiments have the reexamination of the role of the critical cytokines in UVB-induced immune suppression, which was investigated previously by blocking antibodies. The characteristics of the suppressor cells responsible for UVB-induced tolerance were determined. The subcellular mechanism of UVB-induced immune suppression was also explained by the induction of apoptotic cells through the Fas and Fas-ligand interaction. The phagocytosis of the apoptotic cells is believed to induce the production of the immune suppressive cytokine like interleukin-10 by macrophages. Therefore, the therapeutic UVB response to a skin disease, such as psoriasis, by the depletion of infiltrating T cells could be considered in the extension line of apoptosis and immune suppression.

Practical Application of Electron Microscopy in Clinical Medicine (전자현미경의 임상적 응용)

  • Kim Chung-Sook
    • Applied Microscopy
    • /
    • v.8 no.1
    • /
    • pp.81-90
    • /
    • 1978
  • Introduction of electron microscope in biological and medical sciences change the concept of functional and morphologic unit of biological phenomena from the cell to subcellular unit, and it formulated the basis for molecular biology and pathology. Until recently, electron microscopy has mainly been applied to basic research works. However, practical clinical application of electron microscopy is being actively tried. The major clinical fields in which electron microscopy is helpful or even essential include viral diseases, metabolic diseases, glomerular diseases and in the identification of certain types of neoplasms. A brief introduction of characteristics of each conditions are made to encourage more active application of electron microscopy in clinical medicine.

  • PDF

BRCA1 Protein Was Not Expressed in a Normal Human Breast Epithelial Cell Type With Stem Cell and Luminal Characteristics

  • Kang, Kyung-Sun;Maki Saitoh;Angelar Cruz;Chan, Chia-Cheng;Cho, Jae-Jin
    • Toxicological Research
    • /
    • v.14 no.2
    • /
    • pp.123-127
    • /
    • 1998
  • BRCA1 is a tumor suppresser gene in familial cases of breast cancer. It has been controversial whether the subcellular localization of BRCA1 is located in nuclei or cytoplasm in normal human breast cells. We found that a p220 protein was expressed in Type II Normal human breast epithelial cells (NHBEC) but not in Type I NHBEC in Western blot analysis using the 17F8 (3A2) antibody. Immunostaining using the same antibody revealed positive staining in nuclei, cytoplasm and perinuclei of Type II cells and negative staining in Type I NHBEC. The p220 protein, however, was expressed in SV40 immortalized Type I NHBEC and tumorigenic cells derived from them after x-ray and neu oncogene treatment. The subcelluar localization was mostly cytoplasmic and punctate in the nuclei. The breast carcinoma cell lines, MCF-7 and T47D, also expressed the p220 protein. Using RT-PCR, we observed the expression of BRCA1 mRNA in both Type I and Type II NHBEC. This result indicated that there might be mechanisms involved in post-translational or translational regulation of BRCA1 gene. It is speculated that the absence of BRCA1 protein expression in Type I NHBEC might playa role in their susceptibility to neoplastic transformation.

  • PDF