The present proposal seeks support to obtain a rebuilt Philips EM300 transmission electron microscope. Several major proposed projects require electron microscopy, but are not adequately supported by existing equipment. These research projects utilize combinations of morphological, morphometric and immunolocalization analyses in conjunction with biochemical or molecular techniques. (1) To study the influence of calcium ions on differentiation in Physarum polycephalum. Exogenous calcium can induce differentiation and differentiation is accompanied by mobilization of internal calcium stores. Morphometric analyses are proposed to quantitate changes in the mitochondrial content of stored calcium granules. (2) To study the genetic mechanisms regulating eggshell formation during oogenesis in Drosophila by visualizing the effects of well defined mutations on eggshell development and to localizing with high resolution the responsible gene products. (3) To study molecular mechanisms involved in regulation of the mitotic process to localizing with high resolution monoclonal antibodies MPM-1 and MPM-2 which recognize a subset of phosphorylated proteins abundant in mitotic cells. (4) To study calcium regulatory pathways in Trypanosoma brucei by immunolocalization. (5) To use morphometric analyses to provide ultrastructural evidence of free radical damage during aging in tissues of the housefly, Musca domestica. (6) To evaluate the basis of immunosuppression in mice infected with Trichonella pseudospiralis by analysis of adrenal mitochondria and lipid storage droplets in the zona fasciculate layer to provide further evidence of a parasite induced hyperadrenal state. (7) To evaluate, morphologically, the effect of mutations on ATPase organization. (8) To identify changes in seminal vesicle composition and structure which accompany the aging process. Emphasis is place upon the distribution and quantity of type IV collagen, laminin, proteoglycans and testosterone.
