[Histochemical studies of human extraocular muscles (author's transl)].
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to T Takuma.
Explore the source record for details and available documents.
The hormonal regulation of trypsin-like esteroprotease synthesis in mouse submandibular gland was studied at the isozyme level. Antiserum to a mixture of two purified esteroproteases precipitated all the esteroproteases in a crude extract of this gland. Measurement of incorporation of [3H]leucine showed that total esteroprotease synthesis was stimulated by both 5 alpha-dihydrotestosterone and triiodothyronine and that the two hormones had synergistic effects. The observed correlation between the increases of synthetic rate and specific activity of this enzyme suggests that the enzyme level is regulated mainly by the rate of enzyme synthesis. Newly synthesized esteroprotease-antibody complexes gave four peaks of radioactivity with esteroprotease activity and one peak without enzyme activity on isoelectric focusing in acrylamide gel containing 8 M urea. The radioactivities of these five peaks were increased similarly by the two hormones separately or in a combination. These results suggest that the actions of androgens and thyroid homrones in esteroprotease synthesis are indistinguishable at the isozyme level.
Explore the source record for details and available documents.
The hormonal requirements for formation of tyrosine aminotransferase (EC 2.6.1.5) in fetal mouse liver were investigated in organ culture using chemically defined medium. The hormones tested were insulin, thyroxine and prednisolone. Prednisolone alone resulted in a two-fold increase in tyrosine amino-transferase activity in explanted liver in hormone-free medium on day 6, and its effect was dose dependent, but neither insulin nor thyroxine alone induced the enzyme. Addition of prednisolone plus thyroxine and prednisolone plus insulin increased the enzyme activity 1.4- and 1.3-fold, respectively, over that of explants with prednisolone alone. These three hormones together had the greatest effect, causing induction of 1.5-fold more activity than that with prednisolone plus insulin or plus thyroxine. The three hormones were not all needed continuously during the culture period: prednisolone and insulin were required during the early part of cultivation and thyroxine during the later part. The effects of these hormones were blocked by actinomycin D or puromycin, suggesting that these hormones increase de novo synthesis of tyrosine aminotransferase. Phase-contrast microscopy showed that prednisolone stimulated liver epithelial cell outgrowth, probably acting with insulin.
The hormonal requirement for functional differentiation of chick embryo pancreas were investigated by using organ cultures in chemically defined medium. The hormones tested were prednisolone, insulin and thyroxine, and the parameters examined were alpha-amylase (EC 3.2.1.1) and chymotrypsinogen (EC 3.4.4.5) activities, and the ultrastructure of the tissues. Addition of prednisolone alone to explants from 14-day-old chicken embryo pancreas for 3 days increased the activities of amylase and chymotrypsinogen in the tissues by 3.4- and 6.6-fold, respectively, those of tissues before cultivation. Neither thyroxine or insulin alone, nor both hormones together affected pancreatic exocrine differentiation. Thyroxine enhanced the effect of prednisolone on both enzymes, but insulin did not. When the explants were cultured in the medium containing all three hormones, maximum enzyme activities were observed; amylase or chymotrypsinogen activity being 7- or 18-fold, respectively, that of tissues before cultivation. But these three hormones were not simultaneously necessary. Morphological differentiation was also observed in explants cultuvated in medium containing these three hormones. These results suggest that glucocorticoids are essential for normal differentiation of chick pancreas during the late fetal period, possibly with insulin and thyroxine, and also support the idea that pancreatic enzymes are controlled separately.
The actions of thyroxine, 5 alpha-dihydrotestosterone and hydrocortisone singly or in combination in enzyme regulation in the submandibular gland were studied in intact and adrenalectomized female mice. 1. Adrenalectomy decreased the activity of trypsin-like esteroprotease (EC 3.4.4.-), and administration of hydrocortisone to adrenalectomized mice restored the activity to normal. 2. Hydrocortisone and thyroxine had synergistic effects in induction of esteroprotease in adrenalectomized mice, but 5 alpha-dihydrotestosterone and hydrocortisone did not have synergistic effects in either intact or adrenalectomized mice. 3. The activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) was not influenced by change in the glucocorticoid level, but was increased by thyroxine and 5 alpha-dihydrotestosterone in both adrenalectomized mice and intact mice.4. Isoelectric focusing in polyacrylamide gel showed that there are three distinct activities of esteroprotease in this gland with isoelectric points of 5.6, 6.2 and 7.3. Both thyroxine and 5 alpha-dihydrotestosterone similarly induced these activities and glucocorticoids did not affected the isozyme patterns induced by the other two hormones.
The effects of 5 alpha-dihydrotestosterone (DHT) and thyroxine (T4) on glucose-6-phosphate dehydrogenase (G-6-PDH) activity in mouse submandibular gland were investigated histochemically. A strong positive histochemical reaction for G-6-PDH was observed in the excretory ducts of untreated male and female mice, with a slight reaction in the basal portion of the convoluted tubules (striated ducts) of males. Administration of DHT to female mice increased G-6-PDH activity specifically in the convoluted tubules. T4 increased the enzyme activity in the tubules more than DHT. The induction of G-6-PDH activity by T4 in adrenalectomized mice suggests that T4 has a direct effect on the submandibular gland.
The serum levels of somatomedin A, determined by radioreceptor assay, were significantly higher in 57 adult patients with chronic renal failure (X = 2.57 +/- 0.12 U/ml) than in healthy subjects (n = 131; X = 0.77 +/- 0.02 U/ml). A positive correlation was found between somatomedin A and creatinine levels (r = 0.33), but somatomedin A levels did not decrease after hemodialysis. A reduction was only observed after successful renal allografting. Immunoreactive somatomedin A was also found to be increased in patients with chronic renal failure (X = 2.61 +/- 0.21 U/ml), whereas low levels were obtained by the chick bioassay. However, after separating the inhibitory factors from the stimulatory factors by gel chromatography at neutral pH, uremic serum was shown to contain increased levels of somatomedin activity. Although all somatomedin A, determined by different techniques, was present in high molecular forms, the elution pattern differed from that seen in patients with acromegaly.
Esteroprotease, an androgen-dependent enzyme of the mouse submandibular gland, was increased by injection of tri-iodothyronine (T3) in mice with testicular feminization (Tfm) which are genetically deficient in androgen receptors. Histochemical and electron microscopic studies also demonstrated increases of RNA and serous-like granules in cells of the convoluted tubules of the gland. These findings suggest that the esteroprotease gene in Tfm mice is normal and that T3 can induced both esteroprotease and serous-like granules independently of androgen.
Explore the source record for details and available documents.
The hormonal requirements for functional differentiation of mouse parotid glands were investigated using organ cultures in chemically defined medium. The hormones tested were insulin, thyroxine and prednisolone, and the parameters examined were alpha-amylase activity and the ultrastructure of the tissue. It is found that most of the amylase in the cultures (80%) was released into the culture medium after 5 days of cultivation. Prednisolone (5 . 10(-3) mg/ml) alone resulted in a 3--4-fold increase in specific activity of amylase (total amylase activity in the medium and culture) over that in its absence, but neither insulin nor thyroxine alone induced the enzyme. Prednisolone plus thyroxine (over 1 . 10(-7) mg/ml) or insulin (over 1 . 10(-3) unit/ml) induced markedly the enzyme, amylase specific activity being as much as 4- or 6-fold that with prednisolone alone. Moreover the enzyme specific activity was dependent on the prednisolone concentration (5 . 10(-7) - 5 . 10(-3) mg/ml) in the presence of thyroxine (1 . 10(-2) mg/ml) or insulin (1 . 10(-2) unit/ml). Morphological differentiation was also observed in explants cultivated in medium containing prednisolone plus thyroxine or insulin. These results suggest that besides glucocorticoids, insulin and thyroxine are involved in increase in amylase activity in mouse parotid glands during the late suckling period.
Effects of hormones on pepsinogen activity in mouse stomach were investigated by enzyme assay and electron microscopy. Administration of hydrocortisone alone to mice on days 5--10 increased the enzyme activity in the stomach to as much as 4.5-fold that of untreated mice and the increase was dose dependent. Thyroxine also evoked precocious differentiation of the stomach. The effects of thyroxine and hydrocortisone were additive. Injections of insulin had little effect when given alone, or in combination with other hormones. Injection of hydrocortisone alone or plus thyroxine also caused morphological differentiation of the chief cells in the stomach mucosa. Administration of thyroxine to mice on days 15--20 induced as much enzyme activity as that induced by hydrocortisone, but neither of these hormones had any effect when injected after day 23. These results suggest that besides hydrocortisone, thyroxine is also involved in differentiation of the stomach in mice for the first 20 days after birth and that the normal increase of pepsinogen activity in the stomach of mice during the late suckling period is brought about by serum glucocorticoids, possibly with thyroxine.
Explore the source record for details and available documents.
Chymotrypsinogen activity in mouse pancreas gradually decreased from birth, reaching the adult level on day 20 after birth. In suckling mice the enzyme activity was decreased to 1/9 of the control value by injection of thyroxine. The activity was not affected by insulin, and was slightly increased, rather than decreased, by daily injection of hydrocortisone. The effect of thyroxine seemed to be direct, not due to modification of adrenal function.
Changes of alpha-amylase activity (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1) in mouse parotid gland and pancreas were investigated during embryonic and postnatal development. Amylase activity in the parotid gland increased from around day 12 and reached the adult level on day 30. On the other hand, the activity in the pancreas increased during the last stage of gestation, decreased after birth, and then gradually increased from around day 15, reaching the adult level on day 35. Precocious differentiation of the parotid gland was induced by injections of hydrocortisone or thyroxine after birth, but these hormones did not have additive effects on the parotid gland. Injection of insulin had little effect when given alone, but suppressed the effects of the other two hormones on the gland. Only hydrocortisone increased the amylase activity in mouse pancreas during postnatal development, the other two hormones causing slight decrease in pancreatic amylase. Adrenalectomy and injection of hydrocortisone affected the parotid gland but not the pancreas of adult mice. These results suggest that hydrocortisone is involved in cytodifferentiations of the parotid gland and pancreas, and in maintenance of the parotid gland. Thyroxine may also be important in differentiation of the parotid gland.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.