Noriko Nishimura

Immunohistochemical localization of metallothionein in the eye of rats

SummaryIn order to elucidate possible physiological roles of metallothionein (MT), we have studied immunohistological localization of MT in the eye of the rat, using an avidin-biotin peroxidase complex method. As a result, strong MT immunostaining was observed in the epithelium of the lens and cornea. In the retina, considerably strong MT immunostaining was observed in the pigment cell layer while the nerve fiber layer and inner plexiform layer showed weak MT staining. Glial cells in the optic nerve were found to have marked MT staining. The present result is consistent with the hypothesis that MT may be involved not only in activation of zinc enzymes and cell proliferation through supply of zinc ions, but also in a protective mechanism in the blood-retina barrier.

Metallothionein mRNA in the testis and prostate of the rat detected by digoxigenin-labeled riboprobe

Metallothionein (MT), a cysteine-rich heavy metal-binding protein, has been considered to play a role in the homeostatic control and detoxification of heavy metals, such as zinc, copper, and cadmium. In the present study, we have utilized a digoxigenin-labeled riboprobe to localize MT mRNA only by bright-field optics in the testis and prostate of the rat. In the rat testis, MT mRNA was found predominantly in primary spermatocytes and also in secondary spermatocytes and spermatids, but not in the spermatogonia, Sertoli cells, and Leydig cells. On the other hand, MT protein was present in these spermatogenic cells as well as in spermatozoa and Sertoli cells. In the prostate, MT mRNA was found predominantly in the epithelium of the dorsolateral lobes, but not in the ventral lobe, which is in agreement with the observed localization of MT protein. The utilization of both in situ hybridization and immunohistochemical staining on the same tissue specimens show MT gene expression in specific cell types in the male genital organs.

POSSIBLE ROLE OF METALLOTHIONEIN IN THE CELLULAR DEFENSE MECHANISM AGAINST UVB IRRADIATION IN NEONATAL HUMAN SKIN FIBROBLASTS

Abstract –The role of metallothionein (MT) in protecting skin cells against UVB irradiation was investigated. Fibroblast strains from normal adult (HS‐K) and neonatal (NB1RGB) human skins as well as keratinocyte strains from human skin (SV40‐HSK) and newborn Balb/c mouse skin (Pam 212) were exposed to UVB irradiation.

Biliverdin initiates the liver regeneration in the rat — A hypothesis

Abstract Biliverdin has been observed to occur in the blood plasma of a 90% hepatectomized rat. It is also shown that the bile pigment induces a rise in mitotic index in the hepatic parenchymal cells of an intact rat at about 30 h and an elevated rate of hepatic DNA synthesis at 26 h after a single intraperitoneal injection. Hemoglobin, bilirubin, hemin, and protoporphyrin exhibited some of the inducing activity at lower degrees. It is hypothesized that biliverdin initiates the liver-cell-multiplication.

Urinary trehalase as an early indicator of cadmium-induced renal tubular damage in rabbit

The significance of urinary trehalase as a possible early indicator of renal disorder was examined using Cd-treated rabbits, which received 1 mg/kg Cd thrice weekly for 3 months subcutaneously. The results showed that urinary trehalase increased significantly from 1 week after treatment, earlier than LAP, ALP, proteinuria and glucosuria, with no changes in plasma trehalase level. A marked decrease in trehalase activity in renal brush border membranes prepared from Cd-treated rabbits was observed. It was also confirmed by immunohistological techniques that Cd treatment resulted in a marked decrease in specific fluorescence compared with controls. Ouchterlony double diffusion analysis demonstrated that urine and renal brush border extracts formed precipitation lines against anti-renal trehalase IgG, indicating that urinary trehalase and renal trehalase had the same antigenicity.Therefore, the facts presented here would suggest that urinary trehalase originated from the renal brush border, indicating its superiority as a diagnostic tool over other indicative enzymes like LAP and ALP in detecting injury to renal proximal tubular cells in the early stage.

Immunohistochemical Localization of Metallothionein in Organs of Rats Treated with either Cadmium, Inorganic or Organic Mercurials

Metallothionein (MT) is a low, molecular, mass protein inducible by heavy metals such as cadmium (Cd), zinc and copper and having high affinity for these metals. In the present study, we have investigated immunohistological localization of metallothionein in the kidney and brain of rats treated with either Cd, inorganic mercury (Hg) or organic Hg.

The mechanism of cadmium-induced lysozyme enhancement in rabbit kidney

The effect of cadmium on the renal lysozyme level was examined by injecting male albino rabbits subcutaneously with 1 mg cadmium/kg body weight three times a week for 1 or 3 months. The lysozyme level in the renal brush border membrane of the cadmium-treated animals was elevated ten-fold. The lysozyme activity in the liver and small intestine tissue homogenates of rabbits was elevated by a 1-month treatment with cadmium, markedly elevated in the kidney, but markedly reduced in the spleen and lungs. Exposure to cadmium for 3 months produced an essentially similar effect on the enzyme level in the tissue, except for the lungs in which the lysozyme level returned to the preinjection level. This marked increase in the lysozyme level in the kidney of cadmium-treated rabbits was confirmed by an indirect immunofluorescent antibody technique. In control animals, intracellular distribution of the enzyme was selectively distributed to only a small number of proximal tubules, with none distributed in the medulla or glomerulus. However, after expose to cadmium, the renal tubules showed strongly positive lysozyme staining. In addition to an increase in intensity of the specific fluorescence, this enzyme was widely distributed not only in the proximal convoluted portion, but also in the straight portion of the proximal tubules, which essentially showed no enzyme activity under normal conditions. The enzyme in these cells was evenly distributed throughout the cytoplasm. The plasma lysozyme level increased immediately after the administration of cadmium, and detectable amounts of the enzyme began to appear in urine from the 3rd week after the first injection, with a 1-week lag after the maximum level of lysozyme in the plasma. This high level of plasma lysozyme, varied two-to fourfold over the control, and lysozymuria continued throughout the experiment. The concentration of cadmium in the renal cortex was 141 μg/g wet tissue at 1 month, and 208 μg at 3 months. In conclusion, the cadmium-induced enhancement of the lysozyme level in the renal cortex may be due primarily to the elevation of the lysozyme level in plasma by cadmium. The enzymatic high net positive charge, characteristic of lysozyme, may contribute greatly to this mechanism. In addition, the excretion of a large amount of lysozyme into the urine observed in a later stage may be due to the concomitant occurrence of leakage from the destroyed tubular cells and reduced tubular reabsorption of filtered enzyme, whereas lysozymuria at an early stage may be solely due to excess amounts of plasma lysozyme.