Metal ion and guanine nucleotide modulations of agonist interaction in G-protein-coupled serotonin<SUB>1A</SUB> receptors from bovine hippocampus

1. The serotonin type 1A (5-HT1A) receptors are members of a superfamily of seven transmembrane domain receptors that couple to GTP-binding regulatory proteins (G-proteins). We have studied the modulation of agonist binding to 5-HT1A receptors from bovine hippocampus by metal ions and guanine nucleotide. 2. Bovine hippocampal membranes containing the 5-HT1A receptor were isolated. These membranes exhibited high-affinity binding sites for the specific agonist [3H]OH-DPAT. 3. The agonist binding is inhibited by monovalent cations Na+, K+, and Li+ in a concentration-dependent manner. Divalent cations such as Ca2+, Mg2+, and Mn2+, on the other hand, show more complex behavior and induce enhancement of agonist binding up to a certain concentration. The effect of the metal ions on agonist binding is strongly modulated in the presence of GTP-&#947; -S, a nonhydrolyzable analogue of GTP, indicating that these receptors are coupled to G-proteins. 4. To gain further insight into the mechanisms of agonist binding to bovine hippocampal 5-HT1A receptors under these conditions, the binding affinities and binding sites have been analyzed by Scatchard analysis of saturation binding data. Our results are relevant to ongoing analyses of the overall regulation of receptor activity for G-protein-coupled seven transmembrane domain receptors

Modulation of antagonist binding to serotonin<SUB>1A</SUB> receptors from bovine hippocampus by metal ions

1. The serotonin1A(5-HT1A) receptors are members of a superfamily of seven transmembrane domain receptors that couple to G-proteins. They appear to be involved in various behavioral and cognitive functions. Although specific 5-HT1Aagonists have been discovered more than a decade back, the development of selective 5-HT1Aantagonists has been achieved only recently. 2. We have examined the modulation of the specific antagonist [3H]p-MPPF binding to 5-HT1Areceptors from bovine hippocampal membranes by monovalent and divalent metal ions. Our results show that the antagonist binding to 5-HT1Areceptors is inhibited by both monovalent and divalent cations in a concentration-dependent manner. This is accompanied by a concomitant reduction in binding affinity. 3. Our results also show that the specific antagonist p-MPPF binds to all available receptors in the bovine hippocampal membrane irrespective of their state of G-protein coupling and other serotonergic ligands such as 5-HT and OH-DPAT effectively compete with the specific antagonist [3H]p-MPPF. 4. These results are relevant to ongoing analyses of the overall modulation of ligand binding in G-protein-coupled seven transmembrane domain receptors

On the equivalence between topologically and non-topologically massive abelian gauge theories

We analyse the equivalence between topologically massive gauge theory (TMGT) and different formulations of non-topologically massive gauge theories (NTMGTs) in the canonical approach. The different NTMGTs studied are St\"uckelberg formulation of (A) a first order formulation involving one and two form fields, (B) Proca theory, and (C) massive Kalb-Ramond theory. We first quantise these reducible gauge systems by using the phase space extension procedure and using it, identify the phase space variables of NTMGTs which are equivalent to the canonical variables of TMGT and show that under this the Hamiltonian also get mapped. Interestingly it is found that the different NTMGTs are equivalent to different formulations of TMGTs which differ only by a total divergence term. We also provide covariant mappings between the fields in TMGT to NTMGTs at the level of correlation function.Comment: One reference added and a typos corrected. 15 pages, To appear in Mod. Phys. Lett.

Role of disulfides and sulfhydryl groups in agonist and antagonist binding in serotonin<SUB>1A</SUB> receptors from bovine hippocampus

1. The serotonin1A (5-HT1A) receptors are members of a superfamily of seven-transmembrane- domain receptors that couple to G-proteins. They appear to be involved in various behavioral and cognitive functions. Mutagenesis and modeling studies point out that the ligand-binding sites in serotonin receptors are located in the transmembrane domain. However, these binding sites are not very well characterized. Since disulfide bonds and sulfhydryl groups have been shown to play vital roles in the assembly, organization, and function of various G-protein-coupled receptors, we report here the effect of disulfide and sulfhydryl group modifications on the agonist and antagonist binding activity of 5- HT1A receptors from bovine hippocampus. 2. DTT or NEM treatment caused a concentration-dependent reduction in specific binding of the agonist and antagonist in 5-HT1A receptors from bovine hippocampal native and solubilized membranes. This is supported by a concomitant reduction in binding affinity. 3. Pretreatment of the receptor with unlabeled ligands prior to chemical modifications indicate that the majority of disulfides or sulfhydryl groups that undergo modification giving rise to inhibition in binding activity could be at the vicinity of the ligand-binding sites. 4. In addition, ligand-binding studies in presence of GTP-&#947;-S, a nonhydrolyzable analogue of GTP, indicate that sulfhydryl groups (and disulfide bonds to a lesser extent) are vital for efficient coupling between the 5-HT1A receptor and the G-protein. 5. Our results point out that disulfide bonds and sulfhydryl groups could play an important role in ligand binding in 5-HT1A receptors

Molecular basis of association of receptor activity-modifying protein 3 with the family B G protein-coupled secretin receptor

The three receptor activity-modifying proteins (RAMPs) have been recognized as being important for the trafficking and function of a subset of family B G protein-coupled receptors, although the structural basis for this has not been well established. In the current work, we use morphological fluorescence techniques, bioluminescence resonance energy transfer, and bimolecular fluorescence complementation to demonstrate that the secretin receptor associates specifically with RAMP3, but not with RAMP1 or RAMP2. We use truncation constructs, peptide competition experiments, and chimeric secretin-GLP1 receptor constructs to establish that this association is structurally specific, dependent on the intramembranous region of the RAMP and TM6 and TM7 of this receptor. There were no observed changes in secretin-stimulated cAMP, intracellular calcium, ERK1/2 phosphorylation, or receptor internalization in receptor-bearing COS or CHO-K1 cells in the presence or absence of exogenous RAMP transfection, although the secretin receptor trafficks normally to the cell surface in these cells in a RAMP-independent manner, resulting in both free and RAMP-associated receptor on the cell surface. RAMP3 association with this receptor was shown to be capable of rescuing a receptor mutant (G241C) that is normally trapped intracellularly in the biosynthetic machinery. Similarly, secretin receptor expression had functional effects on adrenomedullin activity, with increasing secretin receptor expression competing for RAMP3 association with the calcitonin receptor-like receptor to yield a functional adrenomedullin receptor. These data provide important new insights into the structural basis for RAMP3 interaction with a family B G protein-coupled receptor, potentially providing a highly selective target for drug action. This may be representative of similar interactions between other members of this receptor family and RAMP proteins

Observing mode-dependent wavelength-to-time mapping in few-mode fibers using a single-photon detector array

Wavelength-to-time mapping (WTM)—stretching ultrashort optical pulses in a dispersive medium such that the instantaneous frequency becomes time-dependent—is usually performed using a single-mode fiber. In a number of applications, such as time-stretch imaging (TSI), the use of this single-mode fiber during WTM limits the achievable sampling rate and the imaging quality. Multimode fiber based WTM is a potential route to overcome this challenge and project a more diverse range of light patterns. Here, we demonstrate the use of a twodimensional single-photon avalanche diode (SPAD) array to image, in a time-correlated single-photon counting (TCSPC) manner, the time- and wavelength-dependent arrival of different spatial modes in a few-mode fiber. We then use a TCSPC spectrometer with a onedimensional SPAD array to record and calibrate the wavelength-dependent and mode-dependent WTM processes. The direct measurement of the WTM of the spatial modes opens a convenient route to estimate group velocity dispersion, differential mode delay, and the effective refractive index of different spatial modes. This is applicable to TSI and ultrafast optical imaging, as well as broader areas such as telecommunications

Notoph Gauge Theory: Superfield Formalism

We derive absolutely anticommuting Becchi-Rouet-Stora-Tyutin (BRST) and anti-BRST symmetry transformations for the 4D free Abelian 2-form gauge theory by exploiting the superfield approach to BRST formalism. The antisymmetric tensor gauge field of the above theory was christened as the "notoph" (i.e. the opposite of "photon") gauge field by Ogievetsky and Palubarinov way back in 1966-67. We briefly outline the problems involved in obtaining the absolute anticommutativity of the (anti-) BRST transformations and their resolution within the framework of geometrical superfield approach to BRST formalism. One of the highlights of our results is the emergence of a Curci-Ferrari type of restriction in the context of 4D Abelian 2-form (notoph) gauge theory which renders the nilpotent (anti-) BRST symmetries of the theory to be absolutely anticommutative in nature.Comment: LaTeX file, 12 pages, Talk delivered at SQS'09 (BLTP, JINR, Dubna

Wave forecasting and monitoring during very severe cyclone Phailin in the Bay of Bengal

Wave fields, both measured and forecast during the very severe cyclone Phailin, are discussed in this communication. Waves having maximum height of 13.54 m were recorded at Gopalpur, the landfall point of the cyclone. The forecast and observed significant wave heights matched well at Gopalpur with correlation coefficient of 0.98, RMS error of 0.35 m and scatter index of 14%. Forecasts were also validated in the open ocean and found to be reliable (scatter index &#60; 15%). The study also revealed the presence of Southern Ocean swells with a peak period of 20-22 sec hitting Gopalpur coast along with the cyclone-generated waves