Incorporation of chromatophores into multi-cellular biosensors Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/3x816q14n

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  • Methodologies that detect biologically active substances have important potential applications for medical diagnostics, drug discovery, and chemical and biological anti-terrorism efforts. The wide spectrum of potential analytes that induce a physiological response dictates that novel techniques be developed to more rapidly screen and characterize agents that are more economical and have greater sensitivity than current practices. The research presented in this dissertation describes the development of a biosensor methodology that utilizes optical changes in naturally pigmented chromatophore cells from fish to detect and measure an array of biochemicals and protein toxins. The chromatophores used in this biosensor were harvested from teleost fish sources and the observed patterns of pigment aggregation and dispersion in response to added chemical modulators were used as a reporter mechanism. Differential responses between chromatophore subtypes were utilized as simple cellular sensors for the detection of cholera toxin and in the study of the calcium signaling requirements in these cells. A multi-cellular biosensor was developed that couples pigmentation changes in erythrophores from the teleost Betta splendens with mammalian nerve cell secretory activity. An apparatus was developed that placed PC12 cells, a neuroendocrine cell line, and erythrophores in adjacent chambers connected by a fluid network that allowed erythrophores to be exposed to effluent from PC12 cells; neurotransmitters secreted from PC12 cells induced pigment aggregation in erythrophores. By analyzing the extent of this erythrophore response, this method was capable of detecting the occurrence of substances that altered neurotransmitter secretion levels. A demonstration of this biosensor is presented that detected the inhibition of neurosecretory activity caused by the pathogenic bacterial toxin botulinum, the causative agent of human botulism.
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