Real-time imaging of single nerve cell apoptosis in retinal neurodegeneration.

Apoptotic nerve cell death is implicated in the pathogenesis of several devastating neurodegenerative conditions, including glaucoma and Alzheimer's and Parkinson's diseases. We have devised a noninvasive real-time imaging technique using confocal laser-scanning ophthalmoscopy to visualize single nerve cell apoptosis in vivo, which allows longitudinal study of disease processes that has not previously been possible. Our method utilizes the unique optical properties of the eye, which allow direct microscopic observation of nerve cells in the retina. We have been able to image changes occurring in nerve cell apoptosis over hours, days, and months and show that effects depend on the magnitude of the initial apoptotic inducer in several models of neurodegenerative disease in rat and primate. This technology enables the direct observation of single nerve cell apoptosis in experimental neurodegeneration, providing the opportunity for detailed investigation of fundamental disease mechanisms and the evaluation of interventions with potential clinical applications, together with the possibility of taking this method through to patients.

[1]  Ming Zhao,et al.  Non-invasive detection of apoptosis using magnetic resonance imaging and a targeted contrast agent , 2001, Nature Medicine.

[2]  D. Zack,et al.  Retinal ganglion cell death in experimental glaucoma and after axotomy occurs by apoptosis. , 1995, Investigative ophthalmology & visual science.

[3]  Alex R. Wade,et al.  A fast, robust pattern recognition asystem for low light level image registration and its application to retinal imaging. , 1998, Optics express.

[4]  P. Doevendans,et al.  Real-time imaging of apoptotic cell-membrane changes at the single-cell level in the beating murine heart , 2001, Nature Medicine.

[5]  H. Quigley,et al.  Number of ganglion cells in glaucoma eyes compared with threshold visual field tests in the same persons. , 2000, Investigative ophthalmology & visual science.

[6]  H. Annoura,et al.  Progressive brain dysfunction following intracerebroventricular infusion of beta1–42-amyloid peptide , 2001, Brain Research.

[7]  Thomas L. Chenevert,et al.  Noninvasive real-time imaging of apoptosis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[8]  C Haanen,et al.  A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. , 1995, Journal of immunological methods.

[9]  R. Quirion,et al.  Alzheimer’s disease and the basal forebrain cholinergic system: relations to β-amyloid peptides, cognition, and treatment strategies , 2002, Progress in Neurobiology.

[10]  R S Harwerth,et al.  Ganglion cell losses underlying visual field defects from experimental glaucoma. , 1999, Investigative ophthalmology & visual science.

[11]  P. Thimister,et al.  Visualization of cell death in vivo with the annexin A5 imaging protocol. , 2002, Journal of immunological methods.

[12]  F. Blankenberg,et al.  Annexin-V imaging for noninvasive detection of cardiac allograft rejection , 2001, Nature Medicine.

[13]  T. Deckwerth,et al.  Temporal analysis of events associated with programmed cell death (apoptosis) of sympathetic neurons deprived of nerve growth factor , 1993, The Journal of cell biology.

[14]  F W Fitzke,et al.  Distribution of fundus autofluorescence with a scanning laser ophthalmoscope. , 1995, The British journal of ophthalmology.

[15]  M. Coroneo,et al.  Pressure related apoptosis in neuronal cell lines , 2000, Journal of neuroscience research.

[16]  T. Oshitari,et al.  Rescue of axotomized retinal ganglion cells by BDNF gene electroporation in adult rats. , 2002, Investigative ophthalmology & visual science.

[17]  L. Wheeler,et al.  Neuroprotection of retinal ganglion cells by brimonidine in rats with laser-induced chronic ocular hypertension. , 2001, Investigative ophthalmology & visual science.

[18]  D. Zack,et al.  Caspase activation and amyloid precursor protein cleavage in rat ocular hypertension. , 2002, Investigative ophthalmology & visual science.

[19]  M. Koury,et al.  Comparative analysis of different methodological approaches to the in vitro study of drug-induced apoptosis. , 1999, The American journal of pathology.

[20]  D. Choi,et al.  Staurosporine-Induced Neuronal Apoptosis , 1995, Experimental Neurology.

[21]  A. Sillito,et al.  Surround suppression in primate V1. , 2001, Journal of neurophysiology.

[22]  C. Meshul,et al.  A rat model of chronic pressure-induced optic nerve damage. , 1997, Experimental eye research.

[23]  H. Quigley,et al.  Translimbal laser photocoagulation to the trabecular meshwork as a model of glaucoma in rats. , 2002, Investigative ophthalmology & visual science.

[24]  S. Sharma,et al.  Programmed cell death of retinal ganglion cells during experimental glaucoma. , 1995, Experimental eye research.

[25]  S. Thanos,et al.  Detection of early neuron degeneration and accompanying microglial responses in the retina of a rat model of glaucoma. , 2002, Investigative ophthalmology & visual science.

[26]  M. Vila,et al.  Neurological diseases: Targeting programmed cell death in neurodegenerative diseases , 2003, Nature Reviews Neuroscience.

[27]  W. Tatton,et al.  Retinal damage after 3 to 4 months of elevated intraocular pressure in a rat glaucoma model. , 2000, Investigative ophthalmology & visual science.

[28]  F. Blankenberg,et al.  Will imaging of apoptosis play a role in clinical care? A tale of mice and men , 2001, Apoptosis.

[29]  C. Reutelingsperger,et al.  Apoptosis as a therapeutic target in acutely ischemic myocardium , 2003, Current opinion in cardiology.

[30]  Cynthia A. Schandl,et al.  Major DNA Fragmentation Is a Late Event in Apoptosis , 1997, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.