Cytoskeleton damage

Fig. 8 Confocal fluorescence microscopy of B50 neuroblastoma cells, (a, b) Immunolabeling of mtHSP70 green) in B50 cells in controls (a) and cisPt-treated cells (b). After cisPt, mitochondria are clustered around the nucleus and form dense masses in the cytoplasm (b). Nuclei are counterstained with Hoechst blue), (c, d) Double immunolabeling of filamentous actin red) and a-tubulin green) in B50 control cells (c) and in 48 h cisPt-treated cells (d). CisPt-induced cytoskeleton damage leads tubulin to reorganize into thick bundles (e) and to disruption of filamentous actin microfilaments and accumulation of depolymerized actin at cell periphery (f).

Figure 20.35 Mechanisms by which external or internal stress leads to cell damage resulting in apoptosis. The stress leads to activation of initiator proteolytic enzymes (caspases) that initiate activation of effector caspases. These enzymes cause proteolytic damage to the cytoskeleton, plasma membrane and DNA. The activation of DNAases in the nucleus results in cleavage of DNA chains between histones that produces a specific pattern of DNA damage which, upon electrophoresis, gives a specific pattern of DNA fragments. The major endproduct of apoptosis are the apoptolic bodies which are removed by the phagocytes.

In the blood, 2.5-3.0 g of hemoglobin iron circulates as a component of the erythrocytes (top right). Over the course of several months, the flexibility of the red blood cells constantly declines due to damage to the membrane and cytoskeleton. Old erythrocytes of this type are taken up by macrophages in the spleen and other organs and broken down. The organic part of the heme is oxidized into bilirubin (see p. 194), while the iron returns to the plasma pool. The quantity of heme iron recycled per day is much larger than the amount resorbed by the intestines. 

The progression of the cell cycle is regulated by interconversion processes, in each phase, special Ser/Thr-specific protein kinases are formed, which are known as cyclin-depen-dent kinases (CDKs). This term is used because they have to bind an activator protein (cyclin) in order to become active. At each control point in the cycle, specific CDKs associate with equally phase-specific cyclins. if there are no problems (e.g., DNA damage), the CDK-cyclin complex is activated by phosphorylation and/or dephosphorylation. The activated complex in turn phosphorylates transcription factors, which finally lead to the formation of the proteins that are required in the cell cycle phase concerned (enzymes, cytoskeleton components, other CDKs, and cyclins). The activity of the CDK-cyclin complex is then terminated again by proteolytic cyclin degradation. 

The major secondary events are changes in membrane structure and permeability, changes in the cytoskeleton, mitochondrial damage, depletion of ATP and other cofactors, changes in Ca2+ concentration, DNA damage and poly ADP-ribosylation, lysosomal destabilization, stimulation of apoptosis, and damage to the endoplasmic reticulum. 

Alterations in the cytoskeleton. The cytoskeleton depends on the intracellular Ca2+ concentration, which affects actin bundles, the interactions between actin and myosin and a-tubulin polymerization. The effect of increases in Ca2+ on the cytoskeletal attachments to the plasma membrane and the role of the cytoskeleton in cellular integrity have already been mentioned (see above). If the cytoskeleton is damaged or disrupted or its function altered by an increase in Ca2+, then blebs or protrusions appear on the plasma membrane (see below). As well as an increase in Ca2+, oxidation of, or reaction with sulfydryl groups, such as alkylation or arylation, for example, may disrupt the cytoskeleton, as thiols... 

This process is an early morphological change in cells often seen in isolated cells in vitro but also known to occur in vivo. The blebs, which appear before membrane permeability alters, are initially reversible. However, if the toxic insult is sufficiently severe and the cellular changes become irreversible, the blebs may rupture. If this occurs, vital cellular components may be lost and cell death follows. The occurrence of blebs may be due to damage to the cytoskeleton, which is attached to the plasma membrane as described above. The cause may be an increase in cytosolic Ca2+, interaction with cytoskeletal proteins, or modification of thiol groups (see below). 

Figure 7.46 The metabolism and toxicity of primaquine. Two metabolites, (1) 6-methoxy-8-hydroxyl am in oqui noline and (2) 5-hydroxyprimaquine, are known to be capable of causing oxidative stress and producing ROS. ROS and the metabolites can be removed by GSH, but when this is depleted, they may damage the red cell cytoskeleton and hemoglobin. Abbreviations ROS, reactive oxygen species GSH, glutathione.

Hydroxyprimaquine (Fig. 7.46) will produce ROS in red cells, causing oxidative injury to the cytoskeleton and the oxidation of hemoglobin. The oxidized hemoglobin will bind via disulfide bridges to cytoskeletal proteins (seen as Heinz bodies under the microscope). This and other damages to the red cell lead to their removal by the spleen, and therefore anemia develops. 5-Hydroxyprimaquine also causes a depletion of GSH and the formation of GSS-protein conjugates. This metabolite is considerably more potent than 6-methoxy-8-hydroxylaminoquinoline. 

MS is an autoimmune disease that attacks the myelin sheath of oligodendrocytes around the neuronal axons. This allows the axonal cytoskeleton to be damaged, bringing about secondary axonal loss and persisting neurological dysfunction. The characteristic pathology is of a lesion or plaque in the CNS white matter, formed by inflammation and demyelination and these can be classified into active, chronic active, or chronic silent plaques [86]. 

Genetic errors in or damage to cytoskeletal components can play a central role in disease including many forms of hemolytic anemia and the Duchenne and Becker muscular dystrophies. Appropriate cell-cell contact (regulated through the cytoskeleton) appears to be critical in preventing cells from becoming cancerous and, indeed, many types of cancerous cell exhibit abnormal cytoskeletons. 

Increased acetaldehyde concentrations in the liver cell lead to severe morphological damage, especially in the mitochondria and the cytoskeleton, and to increased lipid peroxidations. 

Intracellular transport of bile acids mainly takes place through the cytoskeleton and intracellular structures (Golgi apparatus, endoplasmic reticulum). Here, too, cholestatic factors can prove to be damaging. Microfilaments are contractile elements not only is the intracellular transport of the bile acids disturbed, but the peristaltic activity of the canaliculi (so-called paralytic cholestasis within the lolsules) is also reduced if the functional capacity of those microfilaments becomes diminished. 

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