Electrical Measurements of Complement-Mediated Membrane Damage in Cultured Nerve and Muscle Cells

Stephens, Cathy Lamar; Henkart, Pierre A.

doi:10.4049/jimmunol.122.2.455

Cited by 32 publications

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Early morphologic alterations in trophically deprived neuronal death in vitro occur without alterations in cytoplasmic Ca2+

Peña

Pilar

2000

J. Comp. Neurol.

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Morphologic and functional parameters altered during neuronal death were investigated in chick ciliary ganglion (CG) neurons in vitro isolated from embryonic day 8 (E8, stage 34). Neurons were separated from nonneuronal cells to investigate their inherent cell death program and were cultured with or without trophic support (choroid, iris, pigment epithelium) from their appropriate target tissue. The cell death process was characterized with investigations focused on the earliest events at the onset of commitment to cell death at 11 hours after plating. Initial morphologic changes in the process of cell death were cytoplasmic; swelling, dendritic retraction, blebbing, vacuolization, which are all characteristics of necrosis. Later, nuclear chromatin condensation occurred, a characteristic of apoptosis. An increase in membrane permeability was measured earlier at 8 hours. During these alterations (associated with the initiation of cell death) single cell analysis was performed to evaluate mobile Ca(2+) changes in the same trophically deprived neurons during the course of the death process; Ca(2+) levels remained at 50 nM. Transient Ca(2+) entry was buffered in control and deprived cells at 13 hours but with different parameters. During the execution stage of death mobile Ca(2+) levels were variable. In this final stage of cell death, neurons demonstrated nuclear damage, cytosol disintegration, or morphology sharing both characteristics. These observations define embryonic CG cell death in vitro as neither purely apoptotic nor necrotic but a form of that exhibits features of both. These results also demonstrate a disassociation of changes in cytosolic Ca(2+) from both CG neuronal survival and trophically deprived cell death in vitro.

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Early morphologic alterations in trophically deprived neuronal death in vitro occur without alterations in cytoplasmic Ca2+

Peña

Pilar

2000

J. Comp. Neurol.

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Restoration of mitochondrial function reverses developmental neuronal death in vitro

Peña

Zhou

Lust

et al. 2001

J of Comparative Neurology

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In a previous study characterizing morphological and functional features of cell death in trophically deprived chick ciliary ganglion neurons (Pena and Pilar [2000] J. Comp. Neurol. 424:377-396), we hypothesized that early cell death events might be targets for reversal, allowing for rescue of dying neurons. To test this hypothesis, ciliary ganglion (CG) neurons were cultured with or without trophic support (choroid, iris, and pigment epithelium soluble extract [CIPE]), or without trophic support for 11 or 18 hours and then exposed to trophic support. Prior to and at the onset of cell death commitment (11 hours) CIPE-deprived neurons exhibited increased membrane permeability, blebbing, cytoplasmic vacuolization, swollen mitochondria, low adenosine triphosphate levels, and release of cytochrome c from mitochondria. CIPE readdition at 11 hours reversed these changes. Between 11 and 18 hours, irreversible DNA fragmentation increased in CIPE-deprived neurons. Cyclosporin A and bongkrekic acid (inhibitors of mitochondrial transition permeability pores) prevented membrane permeability increases and delayed the progression to death in trophically deprived neurons by 12 hours; however, by 48 hours all neurons had died. BOC-Asp-CH2F (BAF), a pan-caspase inhibitor, did not prevent early events of cell death including increased membrane permeability and Cyto c release, but it inhibited DNA fragmentation and prolonged neuronal survival to 48 hours. We conclude that mitochondria changes occur early, prior to commitment and that the suppression of these changes can prevent all the downstream events of death, whereas caspase inhibitors have no effect on the early mitochondria/plasma membrane changes. Mitochondria thus play a critical role in the transition from reversible to irreversible commitment to developmental neuronal death. Furthermore, neuronal death is brought about by activation of one of two distinct pathways, one localized in mitochondria and the other dependent on activation of caspases.

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Interaction of high‐density lipoprotein with Trypanosoma brucei: Effect of membrane stabilizers

Rifkin

1983

J of Cellular Biochemistry

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The specific lysis of bloodstream trypanosomes by serum from a nonpermissive mammalian host is the result of interaction between the serum trypanocidal factor (high-density lipoprotein) and the trypanosome surface. The studies described in this paper attempt to define further the mode of action of this cytotoxic lipoprotein. The binding of high-density lipoprotein to Trypanosoma brucei was instantaneous at 4 degrees C and readily reversible. Binding was not mediated by the surface glycoprotein as removal of the surface coat enhanced binding at 4 degrees C, and no stable glycoprotein-lipoprotein complex could be detected. Pretreatment of trypanosomes with the cross linker dimethylsuberimidate rendered cells resistant to lysis. Addition of membrane-stabilizing drugs, such as cytochalasins C, D, and E, and local anesthetics (dibucaine, tetracaine, and procaine), also inhibited high-density lipoprotein-induced cell lysis. The data presented support the idea that at 37 degrees C lateral diffusion of the variant surface glycoprotein, an integral membrane protein, allows maximal high-density lipoprotein-cell interaction in serum-sensitive cells, and that altered properties of the plasma membrane induced by low temperature or the addition of cytochalasins, local anesthetics, or zinc inhibit this interaction, possibly by increasing the shielding of the plasma membrane by more rigidly anchored surface glycoprotein molecules.

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Electrical Measurements of Complement-Mediated Membrane Damage in Cultured Nerve and Muscle Cells

Cited by 32 publications

References 0 publications

Early morphologic alterations in trophically deprived neuronal death in vitro occur without alterations in cytoplasmic Ca2+

Early morphologic alterations in trophically deprived neuronal death in vitro occur without alterations in cytoplasmic Ca2+

Restoration of mitochondrial function reverses developmental neuronal death in vitro

Interaction of high‐density lipoprotein with Trypanosoma brucei: Effect of membrane stabilizers

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