An algorithm for neurite outgrowth reconstruction

We present a numerical method which provides the ability to analyze digitized microscope images of retinal explants and quantify neurite outgrowth. Few parameters are required as input and limited user interaction is necessary to process an entire experiment of images. This eliminates fatigue related errors and user-related bias common to manual analysis. The method does not rely on stained images and handles images of variable quality. The algorithm is used to determine time and dose dependent, in vitro, neurotoxic effects of 1 GeV per nucleon iron particles in retinal explants. No neurotoxic effects are detected until 72 h after exposure; at 72 h, significant reductions of neurite outgrowth occurred at doses higher than 10 cGy.

[1]  T. Ebendal,et al.  A method for explantation of selected areas of the neural retina , 1986, The Anatomical record.

[2]  Arne Møller,et al.  A simple procedure for quantification of neurite outgrowth based on stereological principles , 2000, Journal of Neuroscience Methods.

[3]  Arne Møller,et al.  A quantitative method for morphometric analysis in neuronal cell culture: unbiased estimation of neuron area and number of branch points , 1995, Journal of Neuroscience Methods.

[4]  J. Bilsland,et al.  A rapid method for semi-quantitative analysis of neurite outgrowth from chick DRG explants using image analysis , 1999, Journal of Neuroscience Methods.

[5]  Jeff W. Lichtman,et al.  Principles of neural development , 1985 .

[6]  David M. Jacobowitz,et al.  A Method for the Rapid Analysis of Neuronal Proportions and Neurite Morphology in Primary Cultures , 1998, Experimental Neurology.

[7]  T. Brümmendorf,et al.  Functional Cooperation of β1-Integrins and Members of the Ig Superfamily in Neurite Outgrowth Induction , 1998, The Journal of Neuroscience.

[8]  T. Ebendal,et al.  Target-field specificity in the induction of retinal neurite outgrowth. , 1987, Brain research.

[9]  Jonathan Bard,et al.  COLLAGEN SUBSTRATA FOR STUDIES ON CELL BEHAVIOR , 1972, The Journal of cell biology.

[10]  T. Ebendal,et al.  Organotypic cultures of neural retina: neurite outgrowth stimulated by brain extracts. , 1983, Brain research.

[11]  T. Matsumoto,et al.  Image analysis of CNS neurotrophic factor effects on neuronal survival and neurite outgrowth , 1990, Journal of Neuroscience Methods.

[12]  J C Sutherland,et al.  Computer network for data acquisition, storage and analysis. , 1997, Journal of photochemistry and photobiology. B, Biology.

[13]  J. Miller,et al.  Detailed characterization of the 1087 MeV/nucleon iron-56 beam used for radiobiology at the alternating gradient synchrotron. , 1998, Radiation research.

[14]  B. Spruijt,et al.  Quantitation of the growth‐associated protein B‐50/GAP‐43 and neurite outgrowth in PC12 cells , 1991, Journal of neuroscience research.

[15]  Bernard W. Agranoff,et al.  A microcomputer-based image analyzer for quantitating neurite outgrowth , 1986, Brain Research.

[16]  G. Vacun,et al.  Organotypic spinal cord culture in serum-free fibrin gel: a new approach to study three-dimensional neurite outgrowth and of neurotoxicity testing Effects of modulating the actin and tubulin dynamics and protein kinase activities , 1997, Journal of Neuroscience Methods.

[17]  Philip H. Gutin,et al.  Radiation Injury to the Nervous System , 1991 .

[18]  E. V. Benton,et al.  Neuritogenesis: a model for space radiation effects on the central nervous system. , 1994, Advances in space research : the official journal of the Committee on Space Research.

[19]  M. Vazquez,et al.  In vitro neurotoxic effects of 1 GeV/n iron particles assessed in retinal explants. , 2000, Advances in space research : the official journal of the Committee on Space Research.

[20]  G. W. Snedecor.  Statistical Methods , 1964 .