| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1991: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1990: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
Morphological and biochemical changes of the connective tissues in amphibian larvae during developmental stages are investigated using immunoflourescence markers and electron microscopy, and compared with the extracellular matrix (ECM) of Anura and Urodela.
In these experiments, the ECM component of the connective tissues in frozen sections of these larvae was stained with antibodies. The antibodies used were as follows : anti-PG-H to the notochordal proteoglycan (PG) of Rana catesbeiana, anti-keratan sulfate PG (KS), anti-chondroitin 4 sulfate PG (C4S), anti-chondroitin 6 sulfate PG (C6S), anti-Type I collagen, anti-Type III collagen, anti-Type IV collagen, and anti-Type VII collagen. In Cynops pyrrhogaster, Type I and Type III collagen antibodies stained the skin and myoseptum, and the peripheral regions containing connective tissues and cartilages. However, Type III collagen antibody was not observed in Rana japonica. Anti-KS especially stained notochord in the early stage of larvae. Positive staining with anti-PG-H of the peripheral regions of the notochordalsheath and ganglia persisted through the late metamorphosis.
Using HPLC, we were able to separate C4S from notochordal PG of Rana catesbeiana for the first time. We are now planning to analyze the core proteins in the PG structure, and apply the results to immunocytochemical studies. The PG distribution and pattern of occurrence may provide us with some clues to help dispel the myths surrounding the evolutional process of these species.
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