Abstract

To understand the precise distribution of tripeptidyl peptidase I (TPP-I), a defect of which has been shown to induce late infantile neuronal ceroid lipofuscinosis, various tissues from rats and mice were analyzed using biochemical and immunohistochemical techniques. Western blot analyses showed that a protein band immunoreactive to anti-TPP-I appeared in tissue extracts of both animals at a molecular weight of approximately 47 kD. Protein levels of TPP-I differed among tissues; they were high in the rat brain, liver, stomach, kidney, thyroid and adrenal glands and in the mouse brain, stomach, kidney, and testis. The proteolytic activity of TPP-I was detectable; it differed in the tissues examined and did not always reflect the expression levels of the protein in the tissues. In particular, the TPP-I activity was low in the brains of both animals and high in the rat testis, although its protein levels were high in the former tissue and low in the latter. Double immunostaining showed the immunoreactivity for TPP-I to be well localized in granular structures of epithelial cells in renal tubules and the cerebral choroid plexus, both of which were also stained with lamp2, a lysosomal membrane protein marker, indicating that TPP-I is a lysosomal enzyme. The immunoreactivity was intense in F4/80-immunopositive macrophages/microglial cells located in various tissues including the thymus, spleen, liver, alimentary tract, and central nervous system. Although the immunoreactivity differed depending on the tissues and even within the same tissues between the species, it was detected in all tissues examined, especially in nerve cells, some types of endocrine cells, and oxyntic cells such as gastric parietal cells and bone osteoclasts. However, the immunoreactivity was faint and week in rat thyroid gland, although its protein level was high in the tissue. These lines of evidence suggest that TPP-I, a lysosomal serine proteinase, is widely distributed in rat and mouse tissues, although its expression levels vary among them.

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