scholarly journals Analysis of Human Mitochondrial DNA Content by Southern Blotting and Nonradioactive Probe Hybridization

2019 ◽  
Vol 80 (1) ◽  
Author(s):  
Joel H. Wheeler ◽  
Carolyn K. J. Young ◽  
Matthew J. Young
Keyword(s):  
Mitochondrial Dna ◽  
Dna Content ◽  
Southern Blotting ◽  
Human Mitochondrial Dna ◽  
Mitochondrial Dna Content ◽  
Probe Hybridization

Development of matured hiPSCs-derived 3D cardiac tissue using ERR gamma agonist and mechanical stress and application for Hypertrophic Cardiomyopathy (HCM) model

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
Y Fujiwara ◽  
K Deguchi ◽  
Y Naka ◽  
M Sasaki ◽  
T Nishimoto ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Hypertrophic Cardiomyopathy ◽  
Mechanical Stress ◽  
Cell Size ◽  
Dna Content ◽  
Disease Model ◽  
Heart Tissue ◽  
Funding Source ◽  
Mitochondrial Dna Content ◽  
Mechanical Stretching

Abstract Introduction Tissue engineering using human induced pluripotent stem cells-derived cardiomyocytes (hiPSCs-CMs) is one of the potential tools to replicate human heart in vitro. Although there are many publications on 3 dimensional (3D) heart tissues (1), these tissues show fetal like phenotypes. For that reason, several maturation methods such as electrical stimulation and mechanical stress have been investigated (2, 3). However, these methods have been inadequate in differentiating fetal like phenotype tissue from adult tissues. Previously, we identified a novel compound, T112, which induced hiPSCs-CMs maturation from approximately 9,000 compounds using Troponin I1-EmGFP and Troponin I3-mCherry double reporter hiPSCs-CMs. This compound enhanced morphological and metabolic maturation of hiPSCs-CMs via estrogen-rerated receptor gamma activation Purpose We hypothesized that our novel compound, T112, in combination with mechanical stress could result in further maturation of 3D heart tissue. Therefore, our specific aim is to develop a novel maturation method applicable to genetic disease model of HCM using 3D heart tissue combined with T112. Methods We constructed 3D heart tissue mixed with fibroblast and double reporter hiPSCs-CMs by the hydrogel methods using Flex cell system®. We added T112 with or without mechanical stretching to 3D tissue from 7 to 15 days after 3D heart tissue was constructed. Then we measured maturation related phenotype such as sarcomere gene expression, mitochondrial DNA content and cell size. Results Similar to hiPSCs-CM, the addition of T112 to the constructed 3D heart tissue significantly increased TNNI3 mRNA compared to that of DMSO. Furthermore, T112 treated 3D heart tissue showed increased cell size and oblong shape. Next, in order to promote more maturation of 3D heart tissue, we performed mechanical stretching with the addition of T112. The combination of T112 with mechanical stretching showed higher expression of mCherry, a reporter protein for TNNI3 expression, and higher isotropy of sarcomere alignment in 3D heart tissue than that with the static condition. Furthermore, 3D heart tissue in the treatment of T112 with or without mechanical stretching showed higher mitochondrial DNA content compared to the respective DMSO controls. Interestingly, we applied this combination method to hiPSCs carrying MYH7 R719Q mutation which is known to cause hypertrophic cardiomyopathy, and the 3D heart tissue composed of cardiomyocytes derived from mutant iPSCs demonstrated increased sarcomere disarray compared to isogenic wild-type 3D heart tissue. Conclusion These results suggest that the combination of T112 and mechanical stretching promotes metabolic and structural maturation of 3D heart tissue and would be useful for creating a HCM disease model. Funding Acknowledgement Type of funding source: Private company. Main funding source(s): T-CiRA project, Takeda Pharmaceutical Company Limited

scholarly journals Low Tumor Mitochondrial DNA Content Is Associated with Better Outcome in Breast Cancer Patients Receiving Anthracycline-Based Chemotherapy

2017 ◽  
Vol 23 (16) ◽  
pp. 4735-4743 ◽  
Author(s):  
Marjolein J.A. Weerts ◽  
Antoinette Hollestelle ◽  
Anieta M. Sieuwerts ◽  
John A. Foekens ◽  
Stefan Sleijfer ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Mitochondrial Dna ◽  
Cancer Patients ◽  
Dna Content ◽  
Breast Cancer Patients ◽  
Mitochondrial Dna Content

Abstract 381: Prolonged Renal Failure Leads to Reduced Number of Active Cardiac Mitochondria in a Rat Model for Long Term Chronic Kidney Disease

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Einat A Hertzberg-Bigelman ◽  
Michal Entin-Meer ◽  
Genya Aharon-Hananel ◽  
Ann Saada ◽  
Ran Levy ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Mitochondrial Dna ◽  
Kidney Disease ◽  
Cardiac Function ◽  
Rat Model ◽  
Dna Content ◽  
Cardiac Mitochondria ◽  
Mitochondrial Content ◽  
Mitochondrial Dna Content

Objectives - Cardiorenal syndrome type 4 is characterized by primary chronic kidney disease (CKD) leading to an impairment of cardiac function. We recently showed a reduced expression of several cardiac mitochondrial genes in short-term CKD rat model. We aimed to evaluate whether cardiac mitochondrial structure and function is modified in long-term CKD and if so, to characterize the potential associated mechanisms. Methods - Lewis rats underwent 5/6 nephrectomy for induction of CKD. Upon necroscopy, eight months later, cardiac sections were analyzed by histology and electron microscopy (EM). Mitochondrial DNA content was determined by the mitochondrial gene, cytochrome B. Mitochondrial content was assessed by citrate synthase (CS) activity in tissue homogenate and respiratory chain function was determined by the activity of complexes I-IV in isolated mitochondria. The levels of PGC1a, a transcription factor for mitochondrial biogenesis, Angiotensin II type 1 receptor and cytosolic cytochrome C were assayed by western blot. Cytokine serum profile was determined by microarray. Results - Long-term CKD leads to cardiac hypertrophy and increased interstitial fibrosis. EM analysis revealed a massive spatial disarrangement accompanied by a considerably increased volume of swollen-damaged mitochondria in CKD hearts (32±3%, n=5, 48±6%, n=4; respectively; p<0.05). Total mitochondrial DNA content was decreased in cardiac tissue of CKD rats. Concomitantly, active mitochondrial content was significantly reduced. Conversely, no differences were observed in respiratory chain enzymes’ functions (complexes I-IV) in isolated active mitochondria. Moreover, inflammatory response and activation of Renin-Angiotensin-Aldosterone-System (RAAS) were detected in the CKD setting. Conclusion - CKD results in a marked reduction of active mitochondria in the heart. Inflammatory cytokines and RAAS, may set a deleterious environment to cardiac mitochondria, as suggested in non-CKD models. The data may represent a significant milestone in the personalized medicine strategy for treating CKD patients who present with normal cardiac function accompanied by positive biomarkers for cardiac mitochondria damage.

Embryo quality, and not chromosome nondiploidy, affects mitochondrial DNA content in mouse blastocysts

2018 ◽  
Vol 234 (7) ◽  
pp. 10481-10488 ◽  
Author(s):  
Ying Jing ◽  
Li Li ◽  
Yuan‐Yuan Li ◽  
Ying‐Chun Ouyang ◽  
Qing‐Yuan Sun ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Dna Content ◽  
Embryo Quality ◽  
Mitochondrial Dna Content

scholarly journals Mitochondrial DNA content and mass increase in progression from normal to hyperplastic to cancer endometrium

2012 ◽  
Vol 5 (1) ◽  
Author(s):  
Antonella Cormio ◽  
Flora Guerra ◽  
Gennaro Cormio ◽  
Vito Pesce ◽  
Flavio Fracasso ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Dna Content ◽  
Mitochondrial Dna Content ◽  
Mass Increase
Sign in / Sign up

Export Citation Format

Share Document