Ultrastructural Changes in Calvarial Osteoblast Cytoskeleton after Prostaglandin E2 Administration in Rats

Recent studies on the cytoskeleton of osteoblasts have been made mainly using cultured cells. However, the morphology of cultured cells may be altered during subculture. Therefore, cytoskeletal changes of calvarial osteoblasts were investigated in situ by electron microscopy using the detergent perfusion method to preserve cell morphology as well as selectively observing the cytoskeleton in the presence of a high concentration of prostaglandin E2 (PGE2) in the calvarial periphery in rats. Rats were perfused with a mixture of Triton X-100 and glutaraldehyde, then the cytoskeleton was observed by transmission electron microscopy. In osteoblasts of the control group, thick bundles of microfilaments ran parallel to the long axis of the cells immediately below the cell membrane adjacent to the osteoid layer. In contrast, in the osteoblasts of the PGE2-administered group, the external morphology was changed to an asteroid or cubic shape, and thick bundles of microfilaments immediately below the cell membrane adjacent to the osteoid were not observed, although microfilament fibres, with a diameter of 5-6 nm, were observed in the cytoplasm.     

Single nucleotide polymorphism genotyping of the barley waxy gene by polymerase chain reaction with confronting two-pair primers

A high‐throughput single nucleotide polymorphism (SNP) genotyping procedure was developed to select amylose‐free barley mutants whose waxy genes had a C‐ to T‐base substitution in exon 5, which converted Gln‐89 of the wild‐type gene into a termination codon. An F₂ population carrying an amylose‐free waxy gene was checked for segregation. Polymerase chain reaction with confronting two‐pair primers (PCR‐CTPP) produced allele‐specific PCR products that have different sizes and are inherited in a co‐dominant manner. Two alleles of the barley waxy gene with SNP were correctly identified in parental strains using the PCR‐CTPP procedure. Segregation of the SNP as detected by PCR‐CTPP in an F₂ population fitted the expected 1:2:1 ratio. The PCR‐CTPP procedure can provide a time saving and cost‐effective alternative to derived cleaved amplified polymorphic sequence in marker‐assisted selection.     

DNA fingerprinting in soybean (Glycine max (L.) Merrill) with oligonucleotide probes for simple repetitive sequences

Summary Soybean DNA fingerprints were analyzed by digoxigenin-labeled oligonucleotide probes complementary to simple repetitive sequences. The clearest and most polymorphic patterns were obtained with (AAT)₆ as a probe, with which all 47 soybean cultivars tested could be distinguished. However, DNA fingerprints of individuals within cultivars showed the same pattern. Using (CT)₈, (GAA)₅ or (AAGG)₄ as probes, clear polymorphic patterns among cultivars and accessions in the subgenus Soja (Glycine max and Glycine soja) were not observed, while quite different patterns were found in accessions in the subgenus Glycine. The results suggest that G. max and G. soja are closer in their genome structure. DNA fingerprints of reciprocal crosses between cultivars and accessions in the subgenus Soja were similar, and contained bands of both parents. In an F₂ population from these crosses, such bands segregated in a Mendelian fashion.     

Endothelin: a novel peptide in the posterior pituitary system

Endothelin (ET), originally characterized as a 21-residue vasoconstrictor peptide from endothelial cells, is present in the porcine spinal cord and may act as a neuropeptide. Endothelin-like immunoreactivity has now been demonstrated by immunohistochemistry in the paraventricular and supraoptic nuclear neurons and their terminals in the posterior pituitary of the pig and the rat. The presence of ET in the porcine hypothalamus was confirmed by reversed-phase high-pressure liquid chromatography and radioimmunoassay. Moreover, in situ hybridization demonstrated ET messenger RNA in porcine paraventricular nuclear neurons. Endothelin-like immunoreactive products in the posterior pituitary of the rat were depleted by water deprivation, suggesting a release of ET under physiological conditions. These findings indicate that ET is synthesized in the posterior pituitary system and may be involved in neurosecretory functions.     

Vertical distributions of two herbivorous cichlid fishes of the genus Tropheus in Lake Tanganyika, Africa

The vertical distributions of two herbivorous cichlid fishes, Tropheus moorii and Tropheus duboisi, were studied on the rocky slope at Luhanga and Bemba, 12 km apart, in Lake Tanganyika, Zaire, Africa. At Luhanga, where T duboisi were not present, T. moorii ranged from the shoreline to a site 30 m deep. At Bemba, T moorii occupied areas less than 18 m and T duboisi areas between 5 and 30 m. In the overlapping zone, 5–18 m depth, each fish held an intra- and interspecific feeding territory, involving size-dependent dominance relationship between neighbors irrespective of species. We indicate that the vertical distributions of these species are restricted by competitive interactions.     

Tetraethylammonium and tetrodotoxin: effects on cochlear potentials

Tetraethylammonium chloride, which is believed to decrease potassium conductance, and tetrodotoxin, which apparently decreases sodium conductance in nerve fibers, were introduced iontophoretically into the organ of Corti or the scala media of guinea pig cochlea. The former depressed the direct-current endocochlear potential and also the alternating-current cochlear microphonics (the receptor potential of the ear), but tetrodotoxin was ineffective except on the nerve impulses.     

Measurement of Si adsorption and “Active Si” in a soil amended with slag fertilizers by using stable isotope 30SP

The silicic acid adsorption by a soil (Eutric Gleysols) where slag fertilizers were applied was measured by the addition of a silicic acid solution labelled with ³⁰Si after soil incubation, in order to study the effect of slag application on the specific Si adsorption by the soil and to estimate the amount of Si in the soil solid phase which can easily enter the soil solution. It was evident that the application of slags increased the ability of soil to adsorb Si. It was also shown that the ³⁰Si added was diluted with not only the Si present in the soil solution but also the Si dissolved from the soil solid phase. We proposed the use of the term “active” for Si in soil which can take part in the isotopic dilution within 1 h. The amount of active Si in the soil solid and liquid phases (D ₆₀ - value) was calculated from the ³⁰Si content in the soil solution and compared with the amount of Si taken up by rice plant (Oryza sativa L.), which was determined in our previous study. The buffering capacity of the soil for Si, which can reflect the ability of soil to maintain the Si concentration in the soil solution constant when Si is added to or removed from the soil, was also determined. The D ₆₀ - value and the Si buffering capacity of the soil increased by slag application. These increases were large when the alkalinity of the applied slags was high. The correlation study revealed that the D ₆₀ - value was a better index of Si availability of the soil than the amount of Si dissolved from the soil solid phase during the incubation when the slags were previously applied.     

Pancreatic insulin secretory response and insulin action in heat-exposed sheep given a concentrate or roughage diet

The effects of heat exposure and type of diet on the insulin secretory response to glucose and glucose disposal in response to insulin action in female sheep were investigated employing hyperglycemic and euglycemic clamp techniques. Animals were divided into concentrate and roughage diet groups, and were maintained at the same intake levels of metabolizable energy and crude protein in both diets. Each diet group was subjected to either thermoneutral (20°C, 70% RH) or hot (30°C, 70% RH) environment, followed by glucose clamp experiments.

Heat-exposed sheep showed significant increases in respiration rates (P<.001) and rectal temperature (P<.05). Plasma glucose concentrations in the basal conditions were lower (P<.01) in the hot environment than in the thermoneutral environment, but there was no significant difference in basal levels of plasma insulin between the environmental treatments. In the hyperglycemic clamp experiment, mean plasma insulin increments increased (P<.05) during the heat exposure period across diet treatments. The ratio of mean plasma insulin increment to glucose infusion rate tended to be higher (P<.07) in the hot environment than in the thermoneutral environment, but diet treatment did not affect the ratio of mean plasma insulin increment to glucose infusion rate. The euglycemic clamp technique showed that glucose infusion rates remained unchanged among treatments. Insulin secretion response to glucose could be stimulated in the hot environment.     

CD38 gene disruption inhibits the contraction induced by alpha-adrenoceptor stimulation in mouse aorta

CD38 is an ectoenzyme with ADP-ribosyl cyclase and hydrolase activities, which synthesizes cyclic ADP-ribose from NAD and hydrolyzes cyclic ADP-ribose to ADP-ribose. It has been shown that cyclic ADP-ribose is a potent Ca(2+) mobilizing messenger in many cells. To know the physiological role of cyclic ADP-ribose in vascular smooth muscle, we examined the effects of various agonists in the aorta isolated from CD38 knockout (CD38(-/-)) mouse. Western blot analysis showed that CD38 protein was detected in the aorta isolated from wild-type (CD38(+/+)) mouse, but not from CD38(-/-) mouse. In the aortae isolated from both CD38(+/+) and CD38(-/-) mice, KCl, phenylephrine and norepinephrine induced concentration-dependent contraction. KCl produced similar concentration-dependent responses in the aortae from both CD38(+/+) and CD38(-/-) mice. Maximum force of contraction induced by KCl (65 mM) was same in the size. Phenylephrine- and norepinephrine-induced contractions were, however, significantly smaller in the aortae from CD38(-/-) mice than in those from CD38(+/+) mice. 5-Hydroxytryptamine, endothelin-1, caffeine and thapsigargin-induced contractions were not significantly different in these two aortae. These results suggest that CD38 gene disruption inhibits alpha-adrenoceptor-induced vascular contractions and cyclic ADP-ribose-mediated signal transduction system is committed in these responses.