Immunohistochemical localization of vasoactive intestinal polypeptide(VIP)-containing neurons in the hypothalamus of the Japanese quail,Coturnix coturnix

Summary The localization of vasoactive intestinal polypeptide (VIP) in the hypothalamus of the quail has been studied by means of light- and electron-microscopic immunohistochemistry. Numerous VIP-immunoreactive perikarya are distributed in the caudal portion of the nucleus infundibularis (n. tuberis) and nucleus mamillaris lateralis, and sparse in the preoptic area, nucleus supraopticus and nucleus paraventricularis. Dense localization of immunoreactive-VIP fibers is observed in the external layer of the median eminence, in close contact with the primary portal capillaries. The main origins of these fiber terminals are VIP-immunoreactive perikarya of the nucleus infundibularis. These neurons are spindle or bipolar and extend one process to the ventricular surface and another to the external layer of median eminence. They are CSF-contacting neurons and apparently constitute the tubero-hypophysial tract that links the third ventricle and the hypophysial portal circulation. VIP-reactive neurons in the nucleus mamillaris lateralis also project axons to the external layer of the median eminence, constituting the posterior bundle of the tuberohypophysial tract. Numerous VIP-immunoreactive perikarya occur also in the nucleus accumbens/pars posterior close to the lateral ventricle. They are also CSF-contacting neurons extending a process to the lateral ventricle. There are moderate distributions of VIP-reactive fibers in the area ventralis and in the area septalis.Ultrastructurally, the immunoreactive products against VIP are found in the elementary granules, 75–115 nm in diameter, within the nerve fibers in the median eminence.This investigation was supported by Scientific Research Grants No. 00556196, No. 56360027 and No. 56760183 from the Ministry of Education of Japan to Professor Mikami and Mr. Yamada     

Phytochrome A-mediated inhibition of seed germination in tomato

Far-red light (FR) inhibition of seed germination of tomato (Solanum lycopersicum L.) was studied with the phytochrome (phy)-hypersensitive mutants, hp-1w, hp-1w,fri1, a phyA-deficient double mutant, and hp-1w,tri1, a phyB1-deficient double mutant. Seeds of all mutants germinated readily in the dark at 25 degrees C, and the germination was retarded by a single 100-s FR pulse given 1-3 h after sowing. The effect of an FR pulse was red-light reversible in all mutants used. After 24 h where a single FR pulse was no longer effective, prolonged FR exposure or hourly FR pulses suppressed germination in hp-1w and hp-1w,tri1, whereas in hp-1w,fri1 the suppressive effect of FR was almost absent. The effect of the prolonged FR was greater than that of the hourly 3-min FR pulses having equal photon fluence, and was fluencerate dependent. Thus we conclude that the germination inhibition by FR in tomato seed consists of a low-fluence response and a high irradiance response (HIR); the latter is controlled by phyA, but not phyB1. This is the first indication of phyA being involved in the HIR of seed germination inhibition.     

Association of the interferon-γ receptor variant (Val14Met) with systemic lupus erythematosus

 Genetic factors seem to play a significant role in susceptibility to systemic lupus erythematosus (SLE). The purpose of this study was to investigate whether the amino acid polymorphism (Val14Met) found within the IFN-γ receptor gene (IFNGR1) plays a prominent role in susceptibility to SLE. We found Val14Met located at the COOH terminal of the signal peptide of the IFN-γ receptor. There was a significant difference in this polymorphism frequency between SLE patients and healthy populations. To clarify whether this amino acid substitution resulted in the alteration of the receptor function, we evaluated the induction of HLA-DR antigen expression on B cells by IFN-γ stimulation. There was also a significant difference in the induction of HLA-DR by IFN-γ stimulation between B cells. Furthermore, an intracellular cytokine assay indicated that the Th1/Th2 balance of Th cells bearing the variant receptor shifted to Th2. The genetic polymorphism found within the IFN-γ receptor gene (Val14Met) may result in a shift to Th2, and this shift may increase susceptibility to SLE. Received: 13 April 1998 / Revised: 30 July 1998     

Propranolol blocks the tachycardia induced by galanin (1-15) but not by galanin (1-29)

The efferent pathways involved in the tachycardia induced by intracisternal injections of the N-terminal galanin fragment (1-15) (GAL (1-15)) and galanin (GAL (1-29)) has been evaluated in rats pretreated with the cholinergic antagonist atropine or the beta-antagonist propranolol. The pretreatment with propranolol significantly blocked the tachycardic and vasopressor effect produced by intracisternal injection of GAL (1-15) (p<0.05), but the pretreatment with atropine did not modify these cardiovascular effects. However, the cardiovascular response elicited by GAL (1-29) is modified by the pretreatment with atropine (p<0.05) but not by propranolol. These findings demonstrate that the central cardiovascular action of GAL (1-15), but not GAL (1-29), is mediated by beta-receptor stimulation and this suggests the existence of a different pathway involved in the cardiovascular response produced by the N-terminal galanin fragment as compared with the parent molecule GAL (1-29).     

Comparative study on the biological properties of 2',5'-oligoadenylate derivatives with purified human RNase L expressed in E. coli

An endoribonuclease, RNase L, which is activated in the presence of 2',5'-linked oligoadenylates, p(1-3)A(2'p5'A)(>2), is the terminal factor of the anti-viral action of interferon. Activation of RNase L results in inhibition of viral proliferation along with induction of apoptosis. Attempts to acquire more effective activators, 2-5A derivatives, have been made for the development of antiviral or anticancer agents. However, the ability of 2-5A derivatives to activate RNase L could not simply be compared due to the diversity of the assay methods used. We have now developed a facile method for assaying the activity of RNase L involving the use of non-fusion RNase L expressed in Escherichia coli and yeast 5S ribosomal RNA as a substrate. Using this method, several 2-5A derivative species have been revaluated. The results suggest that 2-5A molecules modified at the 8-position of the third (from the 5' terminus) adenine ring cause effective dimerization of RNase L and thus increase the ability of RNase L activation.     

Syntheses and receptor-binding studies of derivatives of the opioid antagonist naltrexone

Naltrexone (1), which is a member of the group of competitive opioid antagonists, shows a strong affinity for mu-receptors and its derivatives have been notable as novel receptor antagonists. In this paper, the preparation of several naltrexone derivatives is described; these were used to investigate the role of the oxygenated functional groups in facilitating binding to a series of the opioid receptors. The derivatives showed affinity for opioid mu-receptors which was similar to that of naltrexone, but these compounds, which had masked hydroxyl functional groups, displayed a moderate activity. These results suggest that every oxygenated functional group in naltrexone (1) plays an important role in binding to the opioid receptor.     

Steady-state force-velocity relation in human multi-joint movement determined with force clamp analysis

To study the force-velocity characteristics of human knee-hip extension movement, a dynamometer, in which force was controlled by a servo system, was developed. Seated subjects pressed either bilaterally or unilaterally a force plate, a horizontal position of which was servo-controlled so as to equalize the measured force and a force command generated by a computer at a time resolution of 2 ms (force clamp). The force command was based on the relation between maximum isometric force and foot position within the range between 70% and 90% of "leg length" (LL: longitudinal distance between the sole of the foot and the hip joint), so that the same force relative to the maximum isometric force was consistently applied regardless of the foot position. By regulating the force according to this function, the force-velocity relation was determined. The force-velocity relation obtained was described by a linear function (n=17, r=-0.986 for 80% LL, r=-0.968 for 85% LL) within a range of force between 0.1 and 0.8F(0) (maximum isometric force). The maximum force extrapolated from the linear regression (F(max)) coincided with F(0) (n=17, F(0)/F(max)=1.00+/-0.09 for 80% LL and 1.00+/-0.20 for 85% LL). Also, the velocity at zero force (V(max)) was obtained from the extrapolation. When compared to the bilateral movements, unilateral movements gave rise to a smaller F(max) but the same V(max), suggesting that V(max) is independent of force and therefore represents the proper unloaded velocity. It is suggested that some neural mechanisms may be involved in the force-velocity relation of the knee-hip extension movement, and make it exhibit a linear appearance rather than a hyperbola.     

Possible involvement of endogenous nociceptin/orphanin FQ in the pain-related behavioral responses induced by its own metabolite, nociceptin/orphanin FQ(14-17)

Nociceptin/orphanin FQ(14-17) (N/OFQ(14-17)) is one of the major fragments that are released from N/OFQ, an endogenous ligand for the opioid receptor like-1 (ORL-1) receptor by endopeptidase 24.11. In the present study, we determined the pharmacological profiles of N/OFQ(14-17) on pain-related behavioral responses in the mouse. Intrathecal (i.t.) administration of N/OFQ(14-17) (5-160 pmol) evoked pain-related behaviors, and these behavioral responses were reduced by i.t. co-administration of an ORL-1 receptor antagonist, [Nphe(1)]N/OFQ(1-13)NH2 (4 pmol). However, in the ligand-binding receptor assay, N/OFQ(14-17) had no affinity for the ORL-1 receptor. Furthermore, i.t. pretreatment with an antiserum against N/OFQ (1:50) diminished the N/OFQ(14-17)-induced pain-related behaviors, suggesting that endogenous N/OFQ is involved in their expression. Therefore, N/OFQ(14-17)-induced pain-related behaviors may be mediated through the release of endogenous N/OFQ in the mouse spinal cord.