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Biomedical subjects

T Nakamoto

Publications and source records attributed to T Nakamoto.

At least 145 records · Page 8Linked to original sources

Changes in endoplasmic reticulum during spermiogenesis in the mouse.

Changes in the endoplasmic reticulum of mouse spermatids during spermiogenesis were examined by scanning electron microscopy, applying the OsO4-DMSO-OsO4 method, which permits 3-dimensional observation of cell organelles. At the same time, the endoplasmic reticulum was stained selectively by the Ur-Pb-Cu method, and 0.5 micron-thick sections were prepared for observation by transmission electron microscopy. The results demonstrated stereoscopically the mode of disappearance of the endoplasmic reticulum. In spermatids of the early maturation phase, the endoplasmic reticulum was of uniform diameter, branched and anastomosed, forming a complicated three-dimensional network throughout the cytoplasm. A two-dimensional net was also noted to have formed just beneath the plasma membrane and about Sertoli cell processes invaginating the spermatid cytoplasm. As spermiogenesis progressed, the spread-out endoplasmic reticulum gradually aggregated to form a condensed, glomerulus-like structure consisting of a very thin endoplasmic reticulum connected to the surrounding endoplasmic reticulum. This structure corresponds to the so-called "radial body". Thus, the endoplasmic reticulum may aggregate, condense, be transformed into a radial body, and be removed from the cytoplasm. The two-dimensional endoplasmic reticulum-net, just beneath the plasma membrane and surrounding processes of Sertoli cells, disappeared in spaces where the three-dimensional endoplasmic reticulum network was scarce. Both the two-dimensional endoplasmic reticulum-net structure and the three-dimensional endoplasmic reticulum network disappeared at the same time, indicating that they may be closely related.

Animals↗

The effects of maternal caffeine intake during pregnancy on mineral contents of fetal rat bone.

Various levels of maternal caffeine ingestion during pregnancy were investigated to determine whether caffeine will affect the mineral contents of the growing bones of fetal rats. On day 8 of gestation, rat dams were fed with a 20% protein diet supplemented with 0.5 mg, 1 mg, or 2 mg caffeine/100 g of dams body weight as an experimental group and the same without caffeine as a control until day 22 of gestation. Fetuses were removed by cesarean section on day 22 and mandibular bones were removed to study the mineral contents of calcium, phosphorus, magnesium, and zinc. Although the mandible weighed more in the 0.5-mg caffeine group as compared to the controls, an additional increase of caffeine resulted in a decrease in weight. All calcium, magnesium, and zinc contents per bone decreased in the 1- and 2-mg caffeine groups as compared to either controls or 0.5-mg caffeine group, whereas phosphorus stayed relatively constant regardless of the different levels of caffeine intake. When data are expressed as per gram of bone tissue, most of the mineral contents among the groups disappeared suggesting that normal growth and development of the fetal bone are impaired as a results of maternal caffeine intake. Caffeine intake during gestation in the present study indicates that different levels of caffeine intake may exert not only different effects on mineral contents in bone development, but also affect the growth of the fetal bone.

Animals↗

Interaction between caffeine intake and nutritional status on growing brains in newborn rats.

The purpose of this study was to determine whether caffeine's effects on the growing brain in suckling pups are modified by the nutritional status of the dams. Upon delivery, 8 randomly selected pups were assigned to each dam. They were divided into four groups; group 1 was fed a 20% protein diet as a control; group 2 was fed a 6% protein diet as a malnourished group; group 3 was pair-fed to group 1, but the 20% protein diet was supplemented with caffeine (2 mg/100 g body weight of dams), and group 4 was pair-fed to group 2 with a 6% protein diet with caffeine. At day 15, pups were killed. Brains were removed, weighted and homogenized. Caffeine content of plasma, brain of the pups and maternal milk in groups 3 and 4 were determined. Brains were analyzed for zinc, alkaline phosphatase activity, DNA, protein, and cholesterol. Body weight and protein content of group 3 were greater than group 1, but the zinc contents and alkaline phosphatase activity of group 3 were less than group 1. DNA and cholesterol contents of group 4 were greater than group 2. Supplementation of caffeine to the maternal diet appeared to have various effects on the growing brains of the suckling pups. Caffeine's effects and nutritional status are closely interrelated.

Animals↗

The effect of prenatal protein-energy malnutrition on collagen metabolism in fetal bones.

We analyzed various biochemical variables of the bones in fetal rats whose dams were protein-energy malnourished. Dams were randomly divided into two groups and fed either a 6% protein diet as a malnourished group or a 20% protein diet as a control, from day 13 of gestation to day 22, when fetuses were removed. Hexosamine and hydroxyproline contents of the calvaria and hexosamine contents of long bones were greater in the malnourished group than in the controls. Sequential extractability of collagen differed among various bones in the malnourished group and controls. The ratio of alpha:beta obtained from SDS-polyacrylamide gel of neutral salt-soluble collagen tended to increase in the long bones and mandible, and decrease in the calvaria and ribs in the malnourished group. Also, the ratio of alpha 1:alpha 2 tended to be lower in the malnourished group than in the control group in all bones. Protein-energy malnutrition during pregnancy has shown to affect biochemical composition of various fetal bones.

Animals↗

Analysis of growth factors in renal cell carcinoma.

Tissue extracts prepared from human renal tissue, renal cell carcinoma and serum-free conditioned media of ACHN cells and A498 cells, cell line originated from human renal cell carcinoma, stimulated DNA synthesis of BALB/c 3T3 cells. The activity (growth factor activity) was significantly higher in renal cell carcinoma than in normal tissues. Radioreceptor assay revealed that the contents of epidermal growth factor and type alpha transforming growth factor in the tissue extracts from renal cell carcinoma and conditioned media from renal cell carcinoma cell lines were below detectable level. Most of growth factor activity of the tissue extracts and conditioned media showed high affinity for heparin-Ultrogel, indicating that the major growth factor activity was due to heparin-binding growth factor(s). In addition, renal cell carcinoma contained growth factor activity for ACHN cells, which did not show specific affinity for heparin-Ultrogel.

Binding, Competitive↗

Dynamic changes in Sertoli cell processes invading spermatid cytoplasm during mouse spermiogenesis.

Studies using thick sections stained by ATPase cytochemistry and scanning electron microscopy were carried out to determine three-dimensional ultrastructural alterations in Sertoli cell processes invading neighboring spermatids during mouse spermiogenesis. Sertoli cell processes start invading spermatid cytoplasm at the acrosomal phase of development and undergo considerable change at the maturation phase of development. At step 14, these processes elongate and begin to branch in the spermatid cytoplasm, and by step 15, they extend in various directions to form a complex of canals that the authors have designated the canal complex. The present observations also clarify that the complicated canal complex undergoes regional modification. At the late stages of maturation, the endoplasmic reticulum has gathered with other cell organelles to form aggregates of endoplasmic reticulum in the vicinity of which invading Sertoli cell processes extensively ramify further into thin tubules that intertwine with each other to form a region of thin tubules. In thin sections, each such region was a complex, consisting of small vesicles and endoplasmic reticulum, and corresponded to what has been defined as a mixed body by Morales and Clermont (Anat. Rec., 203:233-244, 1982). During the course of the formation of the region, the invading Sertoli cell processes are continuous at all times with the cell body of the surrounding Sertoli cell.

Adenosine Triphosphatases↗

Proliferation of epithelial cells derived from rat dorsolateral prostate in serum-free primary cell culture and their response to androgen.

Primary cultured epithelial cells derived from the rat dorsolateral prostate proliferated in serum-free nutrient medium WAJC 404 supplemented with mitogens: insulin (650 nM), cholera toxin (120 pM), epidermal growth factor (EGF) (2.5 nM), dexamethasone (300 nM), and bovine pituitary extract (25 micrograms/ml). The culture consisted of two types of epithelial cell colonies: one originated from single cells or small cell aggregates and the other was epithelial cell outgrowth from small tissue fragments attached to a substratum. There were differences in requirements for the mitogens between the two types of colonies. Requirements for cholera toxin, bovine pituitary extract, and dexamethasone were higher in the former type of colonies, and those for EGF were higher in the latter type of colonies. Proliferation of the epithelial cells in either type of colony was suppressed more than 50% by 1 nM dihydrotestosterone. This suppressive effect was not mediated by stromal component in the tissue fragments, and was counteracted by cyproterone acetate, indicating specific and direct action of the androgen on prostate epithelial cells. The results suggest that there is discrete participation of polypeptide growth factors and androgen in proliferation and differentiation, respectively, of prostate epithelial cells in vivo.

Androgens↗

Effects of different levels of caffeine supplemented to the maternal diet on the brains of newborn rats and their dams.

At birth, dams with 8 randomly assigned pups were divided into three groups. Dams of group 1 were fed a control diet. Dams of groups 2 and 3 were fed the control diet supplemented with caffeine (1 mg and 2 mg/100 g body weight, respectively). Pups were killed at day 15 and their brains removed. After weighing, brains were analyzed for DNA, protein, cholesterol, zinc and alkaline phosphatase activity. Brain and plasma caffeine levels were also determined on groups 2 and 3. The dams were milked to measure caffeine levels. The brains from the dams were analyzed for the same parameters as the pups. Caffeine levels in group 3 were consistently higher than in group 2. In the pups, body and brain weights were heavier in group 3 than in the controls. Protein and cholesterol concentrations in group 2 were less than either controls or group 3. Alkaline phosphatase activity in group 2 was higher than either controls or group 3. In the dams, DNA concentration in groups 2 and 3 was less than the controls. Protein and cholesterol concentration in group 2 was less than group 3. It was concluded that low levels of caffeine in the maternal diet during lactation could affect various parameters in the newborn brain. These effects were different from those when the dietary caffeine level was doubled. In contrast, the effects of caffeine on brains of the dams were relatively minor.

Alkaline Phosphatase↗

The influence of a high-protein, low-carbohydrate diet on bone development in the fetuses of rat dams with streptozotocin-induced diabetes.

1. The purpose of the present study was to determine the effects of diet on the mandibles and growth centres of the long bones in the fetuses of diabetic rat dams given a normal diet compared with those given a high-protein, low-carbohydrate diet. 2. On the 9th day of gestation, the controls, groups 1 and 3, were injected with citrate buffer and given 200 and 600 g protein/kg diets respectively. Groups 2 and 4 were injected with 40 mg streptozotocin/kg body-weight and pair-fed with groups 1 and 3 respectively on the 200 and 600 g protein/kg diets. 3. On day 22, some dams were injected with either 45Ca or [14C]proline. Mandibles and long bones were removed and weighed and analysed for Ca content, 45Ca uptake, collagen and collagen synthesis. 4. The body-weights, and mandibular and long-bone weights of the fetuses in the diabetic 200 g protein/kg group were smaller than those of the non-diabetic 200 g protein/kg group, whereas those of the diabetic 600 g protein/kg group showed no difference from the non-diabetic 600 g protein/kg group. 5. The rate of collagen synthesis was higher in the fetuses of the diabetic 600 g protein/kg group than those of the non-diabetic group. Bones of the diabetic 200 g protein/kg group were lower in collagen content when compared with the non-diabetic group, whereas there was no difference between the diabetic and non-diabetic 600 g protein/kg groups.(ABSTRACT TRUNCATED AT 250 WORDS)

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Prenatal protein-energy malnutrition alters various biochemical components of the membranous bones in fetal rats.

The effects of prenatal protein-energy malnutrition on the biochemical parameters of the membranous bone were studied using fetal rats. Timed pregnant rats were fed a protein-deficient diet as an experimental group from day 13 of gestation, whereas control dams were fed a normal protein diet. By day 15, radioactive Na2SO4 was injected. On day 22, all fetuses were delivered by cesarean section. The hexosamine content per milligram dry tissue, and the protein and hexosamine contents per guanidine-HCl extract were greater in the mandibles but less in the calvaria of the malnourished group than in those of the controls. Calcium content per gram dry tissue was lower in both bones of the malnourished group. 35S-sulfate uptake per milligram dry tissue or milligram proteoglycan was greater in the malnourished group than in the controls in both bones. The mandible in the malnourished group had less lower-weight molecular proteoglycan subunits in the dissociative condition. Protein-energy malnutrition affects the mandible and calvaria in different ways, although both bones originate from membranous bone. Insufficient degradation of proteoglycan could be the reason for the delay of mineralization in the malnourished bones.

Animals↗

Chronic caffeine intake alters the composition of various parts of the brain in young growing rats.

Time-pregnant rats were fed a regular laboratory diet until delivery. Litters delivered within an 8-hour period were combined and 8 randomly selected pups were assigned to each dam. Dams were then divided into two groups. Group I received a 20% protein diet. Group II was pair-fed to group I with a 20% protein diet containing caffeine (1 mg/100 g body weight) until day 22 postpartum. On weaning (day 22), only male pups were selected and fed the 20% protein or 20% protein + caffeine diet according to which diet their respective dams had been fed. On day 43, rats were killed and the brains were divided into 6 different areas. DNA, RNA, protein and cholesterol contents were measured. Caffeine supplementation resulted in a decrease in the total brain weight, and DNA and RNA content. DNA value of cortex-midbrain was smaller in the caffeine group whereas the values of medulla oblongata and striatum were greater in comparison with the controls. Caffeine's effects on RNA included an increase in the cerebellum, cortex-midbrain and hippocampus and a decrease in the medulla oblongata. Protein content of cerebellum was increased and of hippocampus decreased due to caffeine supplementation. Cholesterol content of the medulla oblongata and hippocampus was less in the caffeine group than in the control group. The present study shows that chronic caffeine intake during rapid periods of growth influences various parts of the brain in entirely different biochemical manners.

Animals↗

Biochemical and physical alterations of bones in newborn rats due to excess methionine administered either by gastric intubation or by maternal milk.

In one series of experiments, methionine dissolved in physiological saline solution was gastric-intubated into newborn rats every other day from 3 to 13 days after birth. In a second series, diets supplemented with methionine were fed daily to lactating dams from one day after birth of the pups. Pups were killed 15 days after birth and the mandible and long bones removed. The weight and 45Ca uptake of long bone of the groups receiving 0.1 g/100 g body weight of methionine were increased compared with either the group receiving no methionine supplement or that given 0.2 g/100 g body weight. The bone effects of high levels of methionine in the maternal diet were more severe than after intragastric administration. Dietary methionine supplementation during the early growth period is necessary for the normal development of bones in newborn rats, but excess intragastric methionine impaired long-bone development and altered collagen synthesis in long bones and mandibles. High maternal methionine impaired mandibles and long bones, and led to decreased linear dimensions.

Animals↗

Effects of excess methionine on collagen metabolism: a study on newborn rat skin.

We studied the biochemical effects of excess methionine intake on the skin of newborn rats. Group 1 pups were intubated with methionine dissolved in 0.1 ml physiological saline solution in the amount of 0.1 g/100 body wt as a control using a gastric needle. Group 2 pups were given 0.2 g/100 g in the same manner as group 1 as an experimental group. They were intubated every other day from day 3 to 13. On Day 15, [14C]proline was injected intraperitoneally into pups and their skin was removed. 14C total and hydroxyproline uptake was examined in the tissue as well as in the sequential extracts. Although excess methionine intake by the pups did not alter the collagen content of their skin, it caused an increase in the content of type III collagen and a decrease in crosslinked collagen. In addition, newly synthesized collagen in the neutral salt extract increased in the excess methionine group, indicating that crosslinked collagen decreases as excess methionine was intubated. The present study demonstrated that excess methionine in the early lactational period altered the nature of the skin collagen of suckling newborn rat pups.

Animals↗

Gestational protein-energy malnutrition affects the composition of developing skins of rat fetuses and their dams.

1. Various biochemical variables of the skins of rat dams and their fetuses in which protein-energy malnutrition was induced during pregnancy were analysed. 2. One group of dams was fed on a 200 g protein/kg diet as a control and the other was fed on a 60 g protein/kg diet as an experimental group. Each group of dams was fed from day 13 of gestation until day 22. 3. Water, protein and hexosamine concentrations of the fetal skins in the malnourished group were greater than those in the control group, whereas in the dams' skins, protein concentration was greater in the malnourished group than in the control group. 4. Extractability of collagen with neutral salt and pepsin showed no difference between the groups in the skins of fetuses and dams. The content of type III collagen in the fetal skin did not differ between the groups, but was increased in the malnourished dams' skins compared with that of the control group. 5. The present study showed that protein-energy malnutrition during pregnancy significantly affects the metabolism of the skin in both fetuses and their dams. Furthermore, the skins of fetuses and dams are structurally altered in different ways by this nutritional stress.

Animals↗