Abstract 15552: Acute Estrogen-GPER1 Stimulation Preserves Mitochondrial Inner Membrane Protein (Mitofilin) From Degradation caused by Ischemia/Reperfusion Stress

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Tanoya L. C Harris ◽  
Bjorn Olde ◽  
Fredrik Leeb-Lundberg ◽  
Jean C. Bopassa
Keyword(s):  
Electron Microscopy ◽  
Spatial Organization ◽  
Ischemia Reperfusion ◽  
Estrogen Treatment ◽  
Mitochondrial Inner Membrane ◽  
Mitochondrial Integrity ◽  
Inner Membrane Protein ◽  
Estrogen Effect ◽  
And Function ◽  
High Resolution Microscopy

Introduction: We recently found that acute estrogen treatment delays the mitochondrial permeability transition pore opening and reduces ROS production after ischemia/reperfusion, suggesting that estrogen promotes mitochondrial integrity. As mitochondrial inner membrane protein (mitofilin) has been found to control mitochondrial cristae morphology and function, we investigated whether estrogen effect on mitochondrial integrity after ischemia/reperfusion involved regulation of mitofilin via G-Protein Coupled Estrogen Receptor1 (GPER1) activation. Methods: Isolated hearts from male WT (C57BL/6NCrL), and GPER1-/- mice were perfused using Langendorff technique, with and without estrogen (40 nM). Hearts were subjected to 20 min global ischemia followed by 10 min reperfusion. Mitochondria were isolated, and 2D-DIGE followed by mass spectrometry was performed. Mitofilin expression was confirmed by Western blot analysis in mitochondrial fractions. Mitofilin distribution in cardiomyocytes, and its spatial organization in single mitochondria were visualized using high resolution microscopy. Electron microscopy was used to observe the state of mitochondrial cristae morphology. Results: Analysis revealded 52 unique proteins of interest, in which mitofilin was identified. Immunoblot analysis confirmed an increased in mitofilin level with estrogen treatment as compared to control in WT but not in GPER1-/-. We found, as observed in non-ischemic myocytes, that mitofilin in estrogen-treated cardiomyocytes was distributed in the peri-membrane and T-tubules, while only peri-membrane mitofilin was more visible in control group. High resolution microscopy showed a better spatial organization of mitofilin in single mitochondria with estrogen treatment compared to control, in which mitofilin was almost absent. Electron microscopy revealded that mitochondrial morphology was preserved with estrogen treatment, as cristae were well organized compared to control, in which cristae were disrupted. Conclusion: These data indicate that estrogen up-regulates mitofilin expression during ischemia/reperfusion. Estrogen effect on mitofilin may contribute to improved mitochondrial integrity and function.

Abstract 100: G-Protein Coupled Estrogen Receptor1 Prevents Mitochondria Inner Membrane Protein (Mitofilin) Degradation after Ischemia/Reperfusion

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Tanoya L Harris ◽  
Bjorn Olde ◽  
Fredrik L Leeb-Lundberg ◽  
Jean C. Bopassa
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Spatial Organization ◽  
Ischemia Reperfusion ◽  
Estrogen Treatment ◽  
Mitochondrial Integrity ◽  
Inner Membrane Protein ◽  
Estrogen Effect ◽  
And Function ◽  
G Protein Coupled

We recently found that estrogen treatment delays the mitochondrial permeability transition pore opening and reduces ROS production after ischemia/reperfusion, suggesting that estrogen promotes mitochondrial integrity. As mitochondrial inner membrane protein (mitofilin) has been found to control mitochondrial cristae morphology and function, we investigated whether estrogen effect on mitochondrial integrity after ischemia/reperfusion involved regulation of mitofilin via activation of G-Protein Coupled Estrogen Receptor1 (GPER1). Isolated hearts from male WT (C57BL/6NCrL), and GPER1-/- mice were perfused using Langendorff technique, with and without estrogen (40 nM). Hearts were subjected to 20 min global ischemia followed by 10 min reperfusion. Mitochondria were isolated, and 2D-DIGE followed by mass spectrometry was performed. Mitofilin expression level was confirmed using Western blot analysis in mitochondrial fractions. Mitofilin distribution was visualized using confocal microscopy in isolated cardiomyocytes, and its spatial organization in mitochondria was imaged using high resolution fluorescence microscopy. Electron microscopy was used to observe mitochondrial cristae morphology. We obtained 52 unique proteins of interest, in which mitofilin was identified. Immunoblot analysis confirmed the increased in mitofilin level with estrogen treatment as compared to control in WT but not in GPER1 KO. Cardiomyocyte images revealed as observed in non-ischemic myocytes that estrogen treatment conserved mitofilin distribution in the peri-membrane and T-tubules, while only peri-membrane mitofilin was more visible in control group. High resolution microscopy showed a better spatial organization of mitofilin in single mitochondria with estrogen treatment compared to control, in which mitofilin was almost absent. Electron microscopy showed that mitochondrial morphology was conserved in estrogen-treated group as cristae were well organized compared to control, in which cristae were disrupted. These data indicate that estrogen action induces regulation of mitofilin expression during ischemia/reperfusion. Estrogen effect on mitofilin may contribute to improved mitochondrial integrity and function.

Use of human autoantibodies and immunoelectron microscopy to study structure and function of the nucleolus and nuclear bodies

1992 ◽  
Vol 50 (1) ◽  
pp. 506-507
Author(s):  
Robert L. Ochs
Keyword(s):  
Electron Microscopy ◽  
Structure And Function ◽  
Nuclear Bodies ◽  
Nuclear Interior ◽  
Coiled Bodies ◽  
Interchromatin Granules ◽  
And Function ◽  
Conventional Electron Microscopy ◽  
Perichromatin Granules ◽  
Perichromatin Fibrils

By conventional electron microscopy, the formed elements of the nuclear interior include the nucleolus, chromatin, interchromatin granules, perichromatin granules, perichromatin fibrils, and various types of nuclear bodies (Figs. 1a-c). Of these structures, all have been reasonably well characterized structurally and functionally except for nuclear bodies. The most common types of nuclear bodies are simple nuclear bodies and coiled bodies (Figs. 1a,c). Since nuclear bodies are small in size (0.2-1.0 μm in diameter) and infrequent in number, they are often overlooked or simply not observed in any random thin section. The rat liver hepatocyte in Fig. 1b is a case in point. Historically, nuclear bodies are more prominent in hyperactive cells, they often occur in proximity to nucleoli (Fig. 1c), and sometimes they are observed to “bud off” from the nucleolar surface.

Crystal Structures of Fragments D and Double-D from Fibrinogen and Fibrin

1999 ◽  
Vol 82 (08) ◽  
pp. 271-276 ◽  
Author(s):  
Glen Spraggon ◽  
Stephen Everse ◽  
Russell Doolittle
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Crystal Structures ◽  
Structure And Function ◽  
X Ray ◽  
Long Period ◽  
And Function

IntroductionAfter a long period of anticipation,1 the last two years have witnessed the first high-resolution x-ray structures of fragments from fibrinogen and fibrin.2-7 The results confirmed many aspects of fibrinogen structure and function that had previously been inferred from electron microscopy and biochemistry and revealed some unexpected features. Several matters have remained stubbornly unsettled, however, and much more work remains to be done. Here, we review several of the most significant findings that have accompanied the new x-ray structures and discuss some of the problems of the fibrinogen-fibrin conversion that remain unresolved. * Abbreviations: GPR—Gly-Pro-Arg-derivatives; GPRPam—Gly-Pro-Arg-Pro-amide; GHRPam—Gly-His-Arg-Pro-amide

scholarly journals Repair cell first, then regenerate the tissues and organs

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Xiao-Bing Fu
Keyword(s):  
Tissue Repair ◽  
Cell Damage ◽  
Ischemia Reperfusion ◽  
Severe Trauma ◽  
Basic Unit ◽  
Basic Process ◽  
Tissue Repair And Regeneration ◽  
Healing Tissue ◽  
Organ Repair ◽  
And Function

AbstractWound healing, tissue repair and regenerative medicine are in great demand, and great achievements in these fields have been made. The traditional strategy of tissue repair and regeneration has focused on the level of tissues and organs directly; however, the basic process of repair at the cell level is often neglected. Because the cell is the basic unit of organism structure and function; cell damage is caused first by ischemia or ischemia-reperfusion after severe trauma and injury. Then, damage to tissues and organs occurs with massive cell damage, apoptosis and even cell death. Thus, how to achieve the aim of perfect repair and regeneration? The basic process of tissue or organ repair and regeneration should involve repair of cells first, then tissues and organs. In this manuscript, it is my consideration about how to repair the cell first, then regenerate the tissues and organs.

Morphological and Biochemical Features Affecting the Antithrombotic Properties of the Aorta in Adult Rabbits and Rabbit Pups

1998 ◽  
Vol 79 (05) ◽  
pp. 1034-1040 ◽  
Author(s):  
E. Nitschmann ◽  
L. Berry ◽  
S. Bridge ◽  
M. W. C. Hatton ◽  
M. Richardson ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Gel Electrophoresis ◽  
Arterial Wall ◽  
Structure And Function ◽  
Wall Structure ◽  
Matrix Structure ◽  
Antithrombotic Activity ◽  
Antithrombin Activity ◽  
And Function ◽  
Biochemical Features

SummaryWe hypothesised that there are important physiologic differences in arterial wall structure and function with respect to antithrombotic activity in the very young (pre-puberty) compared to adults. Electron microscopy, gel electrophoresis, and activity assays were used to examine differences in aorta structure and function comparing prepubertal rabbits (pups) to adult rabbits. Differences in endothelial function, extracellular matrix structure, proteoglycan (PG) distribution and glycosaminoglycan (GAG) content and function were shown. In both intima and media, total PG, chondroitin sulfate (CS) PG and heparan sulfate (HS) PG content were significantly increased in pups compared to adult rabbits. These findings corresponded to increased concentrations by mass analyses of CS GAG and DS GAG in aortas from pups. There was also a significant increase in antithrombin activity in pups due to HS GAG. In conclusion, differences in both structure and antithrombin activity of aortas from pups compared to adult rabbits suggest that young arteries may have greater antithrombotic potential that is, at least in part, related to increased HS GAG.

scholarly journals Tunable assembly of amyloid-forming peptides into nanosheets as a retrovirus carrier

2015 ◽  
Vol 112 (10) ◽  
pp. 2996-3001 ◽  
Author(s):  
Bin Dai ◽  
Dan Li ◽  
Wenhui Xi ◽  
Fang Luo ◽  
Xiang Zhang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Rational Design ◽  
Structural Model ◽  
Amyloid Fibril ◽  
Computational Approaches ◽  
Retroviral Transduction ◽  
And Function

Using and engineering amyloid as nanomaterials are blossoming trends in bionanotechnology. Here, we show our discovery of an amyloid structure, termed “amyloid-like nanosheet,” formed by a key amyloid-forming segment of Alzheimer’s Aβ. Combining multiple biophysical and computational approaches, we proposed a structural model for the nanosheet that is formed by stacking the amyloid fibril spines perpendicular to the fibril axis. We further used the nanosheet for laboratorial retroviral transduction enhancement and directly visualized the presence of virus on the nanosheet surface by electron microscopy. Furthermore, based on our structural model, we designed nanosheet-forming peptides with different functionalities, elucidating the potential of rational design for amyloid-based materials with novel architecture and function.

Abstract 4336: Interrogating the Human Myocardial Interstitium during Myocardial Arrest and Reperfusion: Dynamic Changes in Cytokines and Protease Activity

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Robert E Stroud ◽  
Christine N Koval ◽  
Isabelle Gengler ◽  
Anne M Deschamps ◽  
John S Ikonomidis ◽  
...  
Keyword(s):  
Detection System ◽  
Ischemia Reperfusion ◽  
Dynamic Assessment ◽  
High Sensitivity ◽  
Fluorescent Detection ◽  
Suspension Array ◽  
Cytokine Analysis ◽  
And Function ◽  
Inflammatory Molecules ◽  
Pulmonary Bypass

Background. Cytokines, such as the interleukins (IL1β, IL2, IL6) and tumor necrosis factor (TNF) can modulate myocardial structure and function with ischemia/reperfusion (I/R) but dynamic assessment of these biological molecules within the human myocardial interstitium with I/R has not been performed, and the inter-relationship to matrix metalloproteinases activity (MMPact) remains unexplored. Accordingly, a fluorogenic microdialysis method was used to simultaneously measure myocardial interstitial cytokine levels and MMPact in patients during and following I/R. Methods . MMPact was measured in patients (n=13) undergoing cardio-pulmonary bypass (CPB) at baseline, during myocardial arrest and CPB (on-CPB), and immediately following reperfusion and separation from CPB (post-CPB) by a validated in-line microdialysis fluorescent detection system. Myocardial interstitial fluid was subjected to cytokine analysis by high sensitivity multiplex suspension array. Results . Interstitial MMPact increased by over 30% post-CPB and was accompanied by a specific change in cytokine profiles (Figure ). The classical pro-inflammatory molecules such as TNF and IL6 were either not detectable or unchanged, whereas IL1β and IL2 which can be proinflammatory, were increased. Conclusions. These unique results demonstrated that a dynamic cytokine signature occurs within the human myocardial interstitium following I/R and is temporally related to heightened MMP activity. Direct interrogation of the human myocardial interstitium may provide a unique insight into critical signaling pathways which may evoke adverse structural and functional events following I/R.

The Tegument and Associated Structures of Fasciola Hepatica

1963 ◽  
Vol s3-104 (68) ◽  
pp. 505-512
Author(s):  
L. T. THREADGOLD
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Light Microscopy ◽  
Fasciola Hepatica ◽  
Golgi Bodies ◽  
New Knowledge ◽  
Pinocytotic Vesicles ◽  
The Individual ◽  
And Function ◽  
Internal Level

The cuticle of light microscopy is shown by electron microscopy to be a surface layer of protoplasm which is an extension of areas of nucleated protoplasm lying deep in the parenchyma. The cuticle therefore exists at two levels. The external level is syncytial, consisting of plateaux separated by branching valleys. This level contains apical pinocytotic vesicles, numerous mitochondria, endoplasmic membranes, large basal and other vacuoles, and dense spines. Tube-like evaginations from the base of the external level connect it to the individual areas of flask-shaped protoplasm which compose the internal level. Each of these areas of protoplasm contains a nucleus, great numbers of mitochondria, some vacuoles and diffuse inclusions, and the Golgi bodies. The histochemistry and function of the cuticle is discussed in the light of this new knowledge of cuticular ultrastructure, and a comparison is made between the cuticle of Cestoda and Trematoda.

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