Assignment On Inclusion Body

Sporadic inclusion-body myositis (s-IBM) is the most common degenerative muscle disease in which aging appears to be a key risk factor. In this review we focus on several cellular molecular mechanisms responsible for multiprotein aggregation and accumulations within s-IBM muscle fibers, and their possible consequences. Those include mechanisms leading to: a) accumulation in the form of aggregates within the muscle fibers, of several proteins, including amyloid-β42 and its oligomers, and phosphorylated tau in the form of paired helical filaments, and we consider their putative detrimental influence; and b) protein misfolding and aggregation, including evidence of abnormal myoproteostasis, such as increased protein transcription, inadequate protein disposal, and abnormal posttranslational modifications of proteins. Pathogenic importance of our recently demonstrated abnormal mitophagy is also discussed. The intriguing phenotypic similarities between s-IBM muscle fibers and the brains of Alzheimer and Parkinson's disease patients, the two most common neurodegenerative diseases associated with aging, are also discussed. This article is part of a Special Issue entitled: Neuromuscular Diseases: Pathology and Molecular Pathogenesis.

Inclusion bodies, sometimes called elementary bodies, are nuclear or cytoplasmic aggregates of stable substances, usually proteins. They typically represent sites of viral multiplication in a bacterium or a eukaryotic cell and usually consist of viral capsid proteins. Inclusion bodies can also be hallmarks of genetic diseases, as in the case of Neuronal Inclusion bodies in disorders like frontotemporal dementia and Parkinson's disease.[1]

Inclusion bodies contain very little host protein, ribosomal components or DNA/RNA fragments. They often almost exclusively contain the over expressed protein and aggregation in inclusion bodies has been reported to be reversible. It has been suggested that inclusion bodies are dynamic structures formed by an unbalanced equilibrium between aggregated and soluble proteins of Escherichia coli. There is a growing body of information indicating that formation of inclusion bodies occurs as a result of intracellular accumulation of partially folded expressed proteins which aggregate through non-covalent hydrophobic or ionic interactions or a combination of both.

Inclusion bodies are dense electron-refractile particles of aggregated protein found in both the cytoplasmic and periplasmic spaces of E. coli during high-level expression of heterologous protein. It is generally assumed that high level expression of non-native protein (higher than 2% of cellular protein) and highly hydrophobic protein is more prone to lead to accumulation as inclusion bodies in E. coli.[citation needed] In the case of proteins having disulfide bonds, formation of protein aggregates as inclusion bodies is anticipated since the reducing environment of bacterial cytosol inhibits the formation of disulfide bonds. The diameter of spherical bacterial inclusion bodies varies from 0.5–1.3 μm and the protein aggregates have either an amorphous or paracrystalline nature depending on the localization.[citation needed] Inclusion bodies have higher density (~1.3 mg ml−1) than many of the cellular components, and thus can be easily separated by high-speed centrifugation after cell disruption. Inclusion bodies despite being dense particles are highly hydrated and have a porous architecture.[2]


Inclusion bodies have a non-unit lipid membrane. Protein inclusion bodies are classically thought to contain misfolded protein. However, this has recently been contested, as green fluorescent protein will sometimes fluoresce in inclusion bodies, which indicates some resemblance of the native structure and researchers have recovered folded protein from inclusion bodies.[3][4][5]

Mechanism of formation[edit]

When genes from one organism are expressed in another organism the resulting protein sometimes forms inclusion bodies. This is often true when large evolutionary distances are crossed: a cDNA isolated from Eukarya for example, and expressed as a recombinant gene in a prokaryote risks the formation of the inactive aggregates of protein known as inclusion bodies. While the cDNA may properly code for a translatable mRNA, the protein that results will emerge in a foreign microenvironment. This often has fatal effects, especially if the intent of cloning is to produce a biologically active protein. For example, eukaryotic systems for carbohydrate modification and membrane transport are not found in prokaryotes. The internal microenvironment of a prokaryoticcell (pH, osmolarity) may differ from that of the original source of the gene. Mechanisms for folding a protein may also be absent, and hydrophobic residues that normally would remain buried may be exposed and available for interaction with similar exposed sites on other ectopic proteins. Processing systems for the cleavage and removal of internal peptides would also be absent in bacteria. The initial attempts to clone insulin in a bacterium suffered all of these deficits. In addition, the fine controls that may keep the concentration of a protein low will also be missing in a prokaryotic cell, and overexpression can result in filling a cell with ectopic protein that, even if it were properly folded, would precipitate by saturating its environment.{citation needed}

Viral inclusion bodies[edit]

Examples of viral inclusion bodies in animals are

Intracytoplasmic eosinophilic (acidophilic)-

Intranuclear eosinophilic (acidophilic)-

Intranuclear basophilic-

Both intranuclear and intracytoplasmic-

Examples of viral inclusion bodies in plants[6] include aggregations of virus particles (like those for Cucumber mosaic virus[7]) and aggregations of viral proteins (like the cylindrical inclusions of potyviruses[8]). Depending on the plant and the plant virus family these inclusions can be found in epidermal cells, mesophyll cells, and stomatal cells when plant tissue is properly stained.[9]

Inclusion bodies in Erythrocytes[edit]

Normally a red blood cell does not contain inclusions in the cytoplasm. However, it may be seen because of certain hematologic disorders.

There are three kinds of erythrocyte inclusions:

  1. Developmental Organelles
    1. Howell-Jolly bodies: small, round fragments of the nucleus resulting from karyorrhexis or nuclear disintegration of the late reticulocyte and stain reddish-blue with Wright stain.
    2. Basophilic stipplings - these stipplings are either fine or coarse, deep blue to purple staining inclusion that appears in erythrocytes on a dried Wright stain.
    3. Pappenheimer bodies - are siderotic granules which are small, irregular, dark-staining granules that appear near the periphery of a young erythrocyte in a Wright stain.
    4. Polychromatophilic red cells - young red cells that no longer have nucleus but still contain some RNA.
    5. Cabot Rings - ring-like structure and may appear in erythrocytes in megaloblastic anemia or in severe anemias, lead poisoning, and in dyserythropoiesis, in which erythrocytes are destroyed before being released from the bone marrow.
  2. Abnormal Hemoglobin Precipitation
    1. Heinz bodies - round bodies, refractile inclusions not visible on a Wright stain film. It is best identified by supravital staining with basic dyes.
    2. Hemoglobin H Inclusions - alpha thalassemia, greenish-blue inclusion bodies appear in many erythrocytes after four drops of blood is incubated with 0.5mL of Brilliant cresyl blue for 20 minutes at 37 °C.
  3. Protozoan Inclusion
    1. Malaria
    2. Babesia

Inclusion bodies in Bacteria[edit]

Polyhydroxyalkanoates or PHA are produced by bacteria as inclusion bodies, the size of PHA granules are limited in E. coli, due to its small bacterial size.[10] Bacterial cell's inclusion bodies are not as abundant intracellularly, in comparison to eukaryotic cells.

Current problems with the isolation of proteins from bacterial inclusion bodies[edit]

70-80% of recombinant proteins expressed E. coli are contained in inclusion bodies (i.e., protein aggregates).[11] The purification of the expressed proteins from inclusion bodies usually require two main steps: extraction of inclusion bodies from the bacteria followed by the solubilisation of the purified inclusion bodies. The use of recombinant proteins can be used to find the mass of misfolding proteins, with the use of mass spectrometry[12]


Pseudo-inclusions are invaginations of the cytoplasm into the cell nuclei, which may give the appearance of intranuclear inclusions. They may appear in papillary thyroid carcinoma.[13]

See also[edit]


  1. ^Cruts M, Gijselinck I, van der Zee J, Engelborghs S, Wils H, Pirici D, Rademakers R, Vandenberghe R, Dermaut B, Martin JJ, van Duijn C, Peeters K, Sciot R, Santens P, De Pooter T, Mattheijssens M, Van den Broeck M, Cuijt I, Vennekens K, De Deyn PP, Kumar-Singh S, Van Broeckhoven C (2006-08-24). "Null mutations in progranulin cause ubiquitin-positive frontotemporal dementia linked to chromosome 17q21". Nature. 442 (7105): 920–4. doi:10.1038/nature05017. PMID 16862115. 
  2. ^Singh, Surinder Mohan; Panda, Amulya Kumar (2005-04-01). "Solubilization and refolding of bacterial inclusion body proteins". Journal of Bioscience and Bioengineering. 99 (4): 303–310. doi:10.1263/jbb.99.303. PMID 16233795. (Subscription required (help)).  
  3. ^Biochem Biophys Res Com 328(2005) 189-197
  4. ^Protein Eng 7(1994) 131-136
  5. ^Biochem Biophys Res Comm 312 (2003) 1383-1386
  6. ^"Plant Viruses Found in Florida and Their Inclusions". University of Florida. Archived from the original on March 24, 2012. 
  7. ^"Inclusions of Cucumber Mosaic Cucumovirus (CMV)". University of Florida. Archived from the original on February 19, 2012. 
  8. ^"Inclusions of Potyviridae Found In Florida". University of Florida. Archived from the original on February 19, 2012. 
  9. ^"Materials and Methods for the Detection of Viral Inclusions". University of Florida. Archived from the original on February 19, 2012. 
  10. ^Jiang XR, Wang H, Shen Chen GQ (2015). "Engineering the bacterial shapes for enhanced inclusion bodied accumulation". Metabolic Engineering. 29: 227–237. doi:10.1016/j.ymben.2015.03.017. 
  11. ^Yang, Zhong, et al. "Highly efficient production of soluble proteins from insoluble inclusion bodies by a two-step-denaturing and refolding method." PloS one 6.7 (2011): e22981.
  12. ^Upadhyay V, Singh A, Panda AK. "Purification of recombinant ovalbumin from inclusion bodies of Escherichia coli". Protein Expression and Purification. 117: 52–58. doi:10.1016/j.pep.2015.09.015. 
  13. ^Chapter 20 in: Mitchell, Richard Sheppard; Kumar, Vinay; Abbas, Abul K.; Fausto, Nelson. Robbins Basic Pathology. Philadelphia: Saunders. ISBN 1-4160-2973-7.  8th edition.
Canine Distemper Virus Cytoplasmic Inclusion Body (Blood smear, Wright's stain)

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