Localization and Interaction of Hyaluronic Acid and Inter-α-Trypsin Inhibitor in Stimulated Preovulatory Rat Ovaries

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Proteins of the inter-α-trypsin inhibitor (IαI) family are reported to stabilize the expanding cumulus extracellular matrix by direct binding with hyaluronic acid in mouse. The aim of this study is to compare the expression and distribution of hyaluronic acid and IαI in the rat ovaries during gonadotropin-induced follicle development. Immuno-and affinity histochemical staining was performed in rat ovary using a polyclonal antibody raised against purified human IαI and a hyaluronic acid-specific probe (biotinylated hyaluronic acid-binding protein). During a preovulatory stage, relatively large amounts of hyaluronic acid were present in the follicular fluid as well as on the surface of expanding cumulus cells and theca/stromal cells. The area of IαI reactivity in stimulated preovulatory follicles was almost the same as that of diffuse deposition of hyaluronic acid. In an in vivo rat model, inhibition of hyaluronic acid synthesis resulted in inhibition of cumulus oocyte complex (COC) expansion accompanied by reduced staining for IαI and hyaluronic acid. Western blot analysis of IαI and immunoassay for hyaluronic acid in tissue extracts from hCG-stimulated ovaries indicated marked increase in IαI and hyaluronic acid expression. These findings support the hypothesis that the co-distribution of IαI and hyaluronic acid acid may serve to organize the formation of the cumulus extracellular matrix in the rat ovary.
日本組織細胞化学会の論文