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Related Concept Videos

Animal Mitochondrial Genetics02:59

Animal Mitochondrial Genetics

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Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
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Blood is specialized connective tissue comprising about 8% of the body mass. It has a thick, liquid extracellular matrix that contains cells, dissolved proteins, and electrolytes, making it five times more viscous than water. Blood is warm, around 38°C, and has an alkaline pH ranging from 7.35 to 7.45.
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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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A eukaryotic cell can have up to three different types of genetic systems: nuclear, mitochondrial, and chloroplast. During evolution, organelles have exported many genes to the nucleus; this transfer is still ongoing in some plant species. Approximately 18% of the Arabidopsis thaliana nuclear genome is thought to be derived from the chloroplast’s cyanobacterial ancestor, and around 75% of the yeast genome derived from the mitochondria’s bacterial ancestor. This export has occurred...
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The human body is composed of cells that are fundamentally made up of several different molecules. These molecules are essential to carry out all physiological processes in the body and are broadly classified into organic and inorganic based on their chemical structures.
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The inner mitochondrial membrane is the primary site of ATP synthesis. The inner membrane domain that forms a smooth layer adjacent to the outer membrane is called the inner boundary membrane. This domain contains membrane transporters that drive metabolites in and out of the mitochondria.  In contrast, the inner membrane network that invaginates into the matrix space is called the cristae membrane. This domain accounts for principle mitochondrial function as it accommodates the protein...
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Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
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Analyzing mitochondrial function in human peripheral blood mononuclear cells.

Chao-Pin Hsiao1, Charles Hoppel2

  • 1Frances Payne Bolton School of Nursing, Case Western Reserve University, USA.

Analytical Biochemistry
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PubMed
Summary

This study details an optimal method for isolating lymphocytes to analyze mitochondrial function and electron transport chain activity. This protocol aids in understanding oxidative phosphorylation disorders using accessible blood samples.

Keywords:
BioenergeticsElectron transport chain complex activityHuman lymphocyteIntegrated mitochondrial functionOxidative phosphorylationPeripheral blood mononuclear cells

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Area of Science:

  • Cellular Biology
  • Mitochondrial Physiology
  • Immunology

Background:

  • Mitochondrial oxidative phosphorylation (OXPHOS) generates cellular energy (ATP).
  • Lymphocytes, abundant in peripheral blood mononuclear cells (PBMCs), are key for monitoring systemic health.
  • Existing research on lymphocyte OXPHOS is limited, particularly regarding electron transport chain (ETC) activity.

Purpose of the Study:

  • To establish an optimal protocol for harvesting human PBMCs.
  • To determine integrated mitochondrial function and analyze ETC complex activity in isolated lymphocytes.
  • To provide a method for investigating OXPHOS disorders using PBMCs.

Main Methods:

  • Harvesting fresh PBMCs from human whole blood using specific anticoagulants and processing media.
  • Utilizing Lymphoprep™ for cell separation and phosphate-buffered saline for washing at room temperature.
  • Assessing integrated mitochondrial function and ETC complex activities in isolated PBMCs.

Main Results:

  • An optimal protocol for PBMC isolation from whole blood was successfully developed.
  • The protocol enables the determination of integrated mitochondrial function.
  • Analysis of electron transport chain complex activities in lymphocytes is feasible with this method.

Conclusions:

  • The presented protocol provides a reliable method for isolating PBMCs for mitochondrial analysis.
  • This technique allows for the comprehensive assessment of lymphocyte mitochondrial function and ETC activity.
  • The findings facilitate the study of OXPHOS defects and related diseases using peripheral blood.