This project seeks to define the inborn biochemical anomalies in the inherited neuro-degenerative diseases of children known as the neuronal ceroid-lipofuscinoses (NCL, Batten disease). It is a continuation of present work using the ovine model and comparative studies, with a change of focus to cellular biology investigations and extends collaboration with Dr. Haltia of Finland specifically investigating the infantile form. This form is distinct from the ovine and late infantile and juvenile forms and others where subunit c of mitochondrial ATP synthase specifically accumulates. The experience and skills learned in the ovine project and its extension of human forms will be applied to the characterization of the storage material in the infantile disease. From the Nature of specifically accumulated species, a strategy will be developed to determine the underlying biochemical lesion. Antibodies will be produced for immunocytochemical studies at light and electron microscopic levels. The ovine work will focus on defining the specific pathway of subunit c turnover, from its synthesis, incorporation into mitochondria and through its normal degradation. Different forms of subunit c storage NCL reflect mutations affecting different associated proteins in this pathway rather than different mutations of a single gene product. Comparative studies with different forms of NCL will help define this pathway. The methodologies will exploit the newly developed radiolabelled E. coli expressed subunit c (with lead sequences) in cell culture and reconstitution studies augmented by characterization of indicated subcellular compartments. Antibodies against different epitopes including lead sequences and mature subunit c as well as markers for particular organelles will be used in immunocytochemical studies. Definition of the underlying defect(s) would have significance for better methods of diagnosis, including prenatal diagnosis and heterozygote detection and may have relevance to therapy. Given the significance of ATP synthase complex, the project will also have relevance to understanding of its assembly and disassembly.
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