Spectrin Proteolysis in the Hippocampus: A Biochemical Marker for Neuronal Injury and Neuroprotection

Roberts-Lewis, Jill M.; Siman, Robert

doi:10.1111/j.1749-6632.1993.tb18290.x

Cited by 48 publications

(34 citation statements)

References 27 publications

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“…Magnification bar equals 100 tim. Taft et al, 1992;Roberts-Lewis and Siman, 1993;Matesic and Lin, 1994;Roberts-Lewis et al, 1994;Kampfl et al, 1996a,b;Posmantur et al, 1996). Several lines of evidence support the hypothesis that cytoskeletal degradation is extremely sensitive to glutamate-and Ca 2~-mediated excitotoxic events.…”

Section: Discussionmentioning

confidence: 87%

“…Several lines of evidence support the hypothesis that cytoskeletal degradation is extremely sensitive to glutamate-and Ca 2~-mediated excitotoxic events. MAP2, spectrin, and neurofilament proteins have been shown to undergo proteolysis after CNS insults involving excitotoxic mechanisms leading to calpain I activation (Banik et al, 1987;Siman and Noszek, 1988;Johnson et al, 1989;Siman et al, 1989;Roberts-Lewis and Siman, 1993;Matesic and Lin, 1994;Roberts-Lewis et al, 1994;Posmantur et al, 1996). In addition, spinal cord injury results in the degradation of neurofilament proteins, which is mediated, in part, by activation of Ca2-dependent proteases (Banik et al, 1982;Iwasaki et aI., 1985Iwasaki et aI., , 1987.…”

Section: Discussionmentioning

confidence: 99%

“…The calpain antibody recognizes the full-length 80-kDa subunit of calpain I, as well as the autolytic 76-kDa form that is generated after activation (Samis et al, 1987;Guttmann et al, 1997). AP-14 recognizes MAP2a and 2b, but not 2c (Binder et al, 1984(Binder et al, , 1986, and Ab 38 is specific for spectrin breakdown products (BPs) resulting from calpain I proteolysis (Roberts-Lewis and Siman, 1993;Roberts-Lewis et al, 1994). It does not recognize full-length a-spectrin, or spectrin BPs produced by other proteases.…”

Section: Antibody Characterizationmentioning

confidence: 99%

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Rapid Calpain I Activation and Cytoskeletal Protein Degradation Following Traumatic Spinal Cord Injury: Attenuation with Riluzole Pretreatment

Springer

Azbill

Kennedy

et al. 1997

Journal of Neurochemistry

129

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Abstract:Immunocytochemical and immunoblotting techniques were used to investigate calpain I activation and the stability of the calpain-sensitive cytoskeletal proteins microtubule-associated protein 2 (MAP2) and spectrin at 1, 4, and 24 h after contusion injury to the spinal cord. Spinal cord injury resulted in the activation of calpain I at all time points examined, with the highest level of activation occurring at 1 h. At the same early time point, there was a loss of dendritic MAP2 staining in spinal cord sections, accompanied by pronounced perikaryal accumulation. The loss in MAP2 staining in the injured spinal cord progressed over the 24-h survival period to affect regions 3 mm distant to the site of injury. The presence of calpain I-specific spectrin degradation was apparent in neuronal cell bodies and fibers as early as 1 h after injury, with the most intense staining occurring within and juxtaposed to the injury site. Spectrin breakdown products in neuronal cell bodies declined rapidly at 4 h and were nearly undetectable at 24 h after injury. Immunoblot studies confirmed the immunocytochemical results by demonstrating a significant increase in calpain I activation, a significant decrease in MAP2 levels, and a significant increase in spectrin breakdown. Finally, treatment of animals with riluzole, an inhibitor of glutamate release, before surgery reduced significantly the loss of MAP2 levels observed at 24 h after injury. These results demonstrate that Ca 2~-dependentprotease activation and degradation of critical cytoskeletal proteins are early events after spinal cord injury and that treatments that minimize the actions of glutamate may limit their breakdown. Key Words: Microtubule-associated protein 2-Spectrin -Cytoskeleton -Glutamate-Calcium -Calpain l-RiluzoIe-CNS injury.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Section: Antibody Characterizationmentioning

confidence: 99%

See 1 more Smart Citation

Rapid Calpain I Activation and Cytoskeletal Protein Degradation Following Traumatic Spinal Cord Injury: Attenuation with Riluzole Pretreatment

Springer

Azbill

Kennedy

et al. 1997

Journal of Neurochemistry

129

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“…Several of these are expressed predominantly or exclusively in neurons, including the β and ζ isoforms of 14-3-3 (Aitken et al, 2003), a hypophosphorylated form of the neurofilament triplet polypeptide NFH (pNFH; Lasek et al, 1985;Jung et al, 2000), and the deubiquitinating enzyme UCH-L1 (Doran et al, 1983). Others are proteolytic fragments of abundant neuronal proteins, including an α-spectrin breakdown product generated by calpain, a family of calcium-dependent proteases associated with neuronal necrosis (Siman et al, 1989;Roberts-Lewis et al, 1993;Zhang et al, 2002). None of these neuron-enriched proteins has been evaluated before as candidate biomarker for the brain and spinal cord damage that can result from surgically-induced deep hypothermic circulation arrest (DHCA; Roche et al, 1996;Watanabe et al, 2004;Selnes and McKhann, 2005).…”

Section: Introductionmentioning

confidence: 99%

Biomarker evidence for mild central nervous system injury after surgically-induced circulation arrest

et al. 2008

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Previously, we identified 14-3-3 β and ζ isoforms and proteolytic fragments of α-spectrin as proteins released from degenerating neurons that also rise markedly in cerebrospinal fluid (CSF) following experimental brain injury or ischemia in rodents, but these proteins have not been studied before as potential biomarkers for ischemic central nervous system injury in humans. Here we describe longitudinal analysis of these proteins along with the neuron-enriched hypophosphorylated neurofilament H (pNFH) and the deubiquitinating enzyme UCH-L1 in lumbar CSF samples from 19 surgical cases of aortic aneurysm repair, 7 involving cardiopulmonary bypass with deep hypothermic circulatory arrest (DHCA). CSF levels of the proteins were near the lower limit of detection by Western blot or enzyme-linked fluorescence immunoassay at the onset of surgical procedures, but increased substantially in a subset of cases, typically within 12-24 hours. All cases involving DHCA were characterized by >3-fold elevations in CSF levels of the two 14-3-3 isoforms, UCH-L1, and pNFH. Six of 7 also exhibited marked increases in α-spectrin fragments generated by calpain, a protease known to trigger necrotic neurodegeneration. Among cases involving aortic cross-clamping but not DHCA, the proteins rose in CSF preferentially in the subset experiencing acute neurological complications. Our results suggest the neuron-enriched 14-3-3β, 14-3-3ζ, pNFH, UCH-L1, and calpain-cleaved α-spectrin may serve as a panel of biomarkers with clinical potential for the detection and management of ischemic central nervous system injury, including for mild damage associated with surgically-induced circulation arrest.

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“…During platelet activation, which includes a cell shape transformation from discs into irregular spheres, spectrin is cleaved to ϳ150-kDa fragments by the calcium-dependent protease, calpain (3). ␣-Spectrin cleavage by calpain has also been implicated in cellular hypoxia (4), neuronal injury and degeneration (5), and neuronal growth cone formation (6). However, in apoptotic cells, ␣-spectrin proteolysis to ϳ150-kDa fragments is mediated by caspases; in these cells, spectrin proteolysis is thought to be important for the disintegration of the plasma membranes via formation of vesicular "apoptotic bodies" (7)(8)(9)(10)(11)(12).…”

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confidence: 99%

Caspase Remodeling of the Spectrin Membrane Skeleton during Lens Development and Aging

Lee

Morrow

Fowler

2001

Journal of Biological Chemistry

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Terminal differentiation of lens fiber cells resembles the apoptotic process in that organelles are lost, DNA is fragmented, and changes in membrane morphology occur. However, unlike classically apoptotic cells, which are disintegrated by membrane blebbing and vesiculation, aging lens fiber cells are compressed into the center of the lens, where they undergo cell-cell fusion and the formation of specialized membrane interdigitations. In classically apoptotic cells, caspase cleavage of the cytoskeletal protein ␣-spectrin to ϳ150-kDa fragments is believed to be important for membrane blebbing. We report that caspase(s) cleave ␣-spectrin to ϳ150-kDa fragments and ␤-spectrin to ϳ120-and ϳ80-kDa fragments during late embryonic chick lens development. These fragments continue to accumulate with age so that in the oldest fiber cells of the adult lens, most, if not all, of the spectrin is cleaved to discrete fragments. Thus, unlike classical apoptosis, where caspase-cleaved spectrin is short lived, lens fiber cells contain spectrin fragments that appear to be stable for the lifetime of the organism. Moreover, fragmentation of spectrin results in reduced membrane association and thus may lead to permanent remodeling of the membrane skeleton. Partial and specific proteolysis of membrane skeleton components by caspases may be important for age-related membrane changes in the lens.The spectrin-actin membrane skeleton underlies the plasma membranes of all cells and is important for cellular shape, membrane stability and deformability, as well as the formation of membrane subdomains (1). The major component of the membrane skeleton, spectrin, is composed of an ␣/␤ heterodimer that self-associates head-to-head to form a 200-nm extended tetramer filament. Spectrin cross-links actin filaments into an isotropic meshwork. This spectrin-actin meshwork is attached to the membrane by direct interactions of ␤-spectrin with membrane proteins and indirect interactions of ␤-spectrin with membrane attachment proteins such as ankyrin (2).Proteolysis of ␣-spectrin (␣II-spectrin, non-erythroid spectrin, or fodrin) to discrete fragments is implicated in changes in cell shape and membrane morphology which occur in many cell types. During platelet activation, which includes a cell shape transformation from discs into irregular spheres, spectrin is cleaved to ϳ150-kDa fragments by the calcium-dependent protease, calpain (3). ␣-Spectrin cleavage by calpain has also been implicated in cellular hypoxia (4), neuronal injury and degeneration (5), and neuronal growth cone formation (6). However, in apoptotic cells, ␣-spectrin proteolysis to ϳ150-kDa fragments is mediated by caspases; in these cells, spectrin proteolysis is thought to be important for the disintegration of the plasma membranes via formation of vesicular "apoptotic bodies" (7-12). Although calpain cleavage of spectrin is known to affect its ability to bind membranes or actin filaments (13, 14), the detailed consequences of caspase cleavage of spectrin have not been studied.The terminal ...

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Spectrin Proteolysis in the Hippocampus: A Biochemical Marker for Neuronal Injury and Neuroprotection

Cited by 48 publications

References 27 publications

Rapid Calpain I Activation and Cytoskeletal Protein Degradation Following Traumatic Spinal Cord Injury: Attenuation with Riluzole Pretreatment

Rapid Calpain I Activation and Cytoskeletal Protein Degradation Following Traumatic Spinal Cord Injury: Attenuation with Riluzole Pretreatment

Biomarker evidence for mild central nervous system injury after surgically-induced circulation arrest

Caspase Remodeling of the Spectrin Membrane Skeleton during Lens Development and Aging

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