Molecular basis for cis-urocanic acid as a 5-HT2A receptor agonist

The underlying mechanisms of urocanic acid (UA) to induce immune suppression remain elusive until the recent finding that cis-UA acts via the serotonin, 5-hydroxytryptamine (5-HT) receptor subtype 5-HT_2A. In the present study, the interactions of cis-UA to 5-HT_2A receptor were explored and compared with those of 5-HT to the same receptor using computational docking. Similar binding modes were observed for cis-UA and 5-HT with 5-HT_2A receptor and the former possessed relatively higher binding affinity, which may account for cis-UA being a serotonin receptor agonist. Moreover, the molecular basis for the distinct binding affinities between the trans- and cis-UA with 5-HT_2A receptor was also provided.     

Bioisosteric replacement of the hydrazide pharmacophore of the cannabinoid-1 receptor antagonist SR141716A. Part I: Potent, orally-active 1,4-disubstituted imidazoles

A new series of CB₁ receptor antagonists incorporating an imidazole-based isosteric replacement for the hydrazide moiety of rimonabant (SR141716) is disclosed. Members of this imidazole series possess potent/selective binding to the rCB₁ receptor and exhibit potent hCB₁ functional activity. Isopropyl analog 9a demonstrated activity in the tetrad assay and was orally-active in a food intake model.     

Molecular modeling of flexible arm‐mediated interactions between bacterial chemoreceptors and their modification enzyme

Sensory adaptation in bacterial chemotaxis is mediated by methylation and demethylation of specific glutamyl residues in the cytoplasmic domain of chemoreceptors. Methylation is catalyzed by methyltransferase CheR. In E. coli and related organisms, methylation sufficiently rapid to be physiologically effective requires a carboxyl terminal pentapeptide sequence on the receptor being modified or, via adaptational assistance, on a neighboring homodimer in a receptor cluster. Pentapeptide‐enhanced methylation is thought to be mediated by a ～30 residue, potentially disordered sequence that serves as a flexible arm connecting the receptor body and pentapeptide‐bound methyltransferase, thus allowing diffusionally restricted enzyme to reach methyl‐accepting sites. However, it was not known how many or which sites on the same or neighboring receptors were accessible to the tethered enzyme. We investigated using molecular modeling and found that, in a hexagonal array of trimers of receptor dimers, CheR tethered to a dimer of chemoreceptor Tar by its native 30‐residue flexible‐arm sequence could reach all methyl‐accepting sites on the dimer to which it was tethered plus 48 methyl‐accepting sites distributed among nine neighboring dimers, equivalent to the total sites carried by six receptors. This modeling‐determined methylation neighborhood of one enzyme‐binding dimer and six neighbors corresponds precisely with the experimentally identified neighborhood of seven. Thus, the experimentally observed adaptational assistance can occur by docking of pentapeptide‐bound, diffusionally restricted enzyme to methyl‐accepting sites on neighboring receptors. Our analysis introduces the notion that physiologically relevant adaptational assistance could occur even if only a subset of sites on a particular receptor are within reach.     

Synthesis, structure–affinity relationships, and modeling of AMDA analogs at 5-HT2A and H1 receptors: Structural factors contributing to selectivity

Histamine H₁ and serotonin 5-HT_2A receptors present in the CNS have been implicated in various neuropsychiatric disorders. 9-Aminomethyl-9,10-dihydroanthracene (AMDA), a conformationally constrained diarylalkyl amine derivative, has affinity for both of these receptors. A structure–affinity relationship (SAFIR) study was carried out studying the effects of N-methylation, varying the linker chain length and constraint of the aromatic rings on the binding affinities of the compounds with the 5-HT_2A and H₁ receptors. Homology modeling of the 5-HT_2A and H₁ receptors suggests that AMDA and its analogs, the parent of which is a 5-HT_2A antagonist, can bind in a fashion analogous to that of classical H₁ antagonists whose ring systems are oriented toward the fifth and sixth transmembrane helices. The modeled orientation of the ligands are consistent with the reported site-directed mutagenesis data for 5-HT_2A and H₁ receptors and provide a potential explanation for the selectivity of ligands acting at both receptors.     

Structural determinants for selective recognition of peptide ligands for endothelin receptor subtypes ETA and ETB

The molecular basis for recognition of peptide ligands endothelin‐1, ‐2 and ‐3 in endothelin receptors is poorly understood. Especially the origin of ligand selectivity for ET_A or ET_B is not clearly resolved. We derived sequence‐structure‐function relationships of peptides and receptors from mutational data and homology modeling. Our major findings are the dissection of peptide ligands into four epitopes and the delineation of four complementary structural portions on receptor side explaining ligand recognition in both endothelin receptor subtypes. In addition, structural determinants for ligand selectivity could be described. As a result, we could improve the selectivity of BQ3020 about 10‐fold by a single amino acid substitution, validating our hypothesis for ligand selectivity caused by different entrances to the receptors' transmembrane binding sites. A narrow tunnel shape in ET_A is restrictive for a selected group of peptide ligands' N‐termini, whereas a broad funnel‐shaped entrance in ET_B accepts a variety of different shapes and properties of ligands. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd.     

Zebrafish β-adrenergic receptor mRNA expression and control of pigmentation

Beta adrenergic receptors (β-ARs) are members of the G-protein-coupled receptor superfamily and mediate various physiological processes in many species. The expression patterns and functions of β-ARs in zebrafish are, however, largely unknown. We have identified zebrafish β-AR orthologs, which we have designated as adrb1, adrb2a, adrb2b, adrb3a and adrb3b. adrb1 was found to be expressed in the heart and brain. Expression of adrb2a predominated in the brain and skin, whereas adrb2b was found to be highly expressed in muscle, pancreas and liver. Both adrb3a and adrb3b were exclusively expressed in blood. Knock-down of these β-ARs by morpholino oligonucleotides revealed a functional importance of adrb2a in pigmentation. Expression of atp5a1 and atp5b, genes that encode subunits of F1F0-ATPase, which is known to be involved in pigmentation, was significantly increased by knock-down of adrb2a. Our data suggest that adrb2a may regulate pigmentation, partly by modulating F1F0-ATPase.     

Pig melatonin receptor 1a (MTNR1A) genotype is associated with seasonal variation of sow litter size

The duration of nocturnal melatonin secretion reaches its minimum in summer, a physiological event that is likely related with the diminished sow fertility and delayed puberty typically observed in this season. Melatonin exerts its function by binding two different receptors named as MTNR1A and MTNR1B. Interestingly, the MTNR1A gene is located on a chromosome SSC17 region where QTL for prolificacy traits have been detected in previous studies. In this work, we have found a synonymous T162C polymorphism at exon 2 of the pig MTNR1A gene. An association analysis between this polymorphism and sow prolificacy in an Iberian ×  Meishan intercross was performed. The utilization of four statistical models of increasing complexity demonstrated that the MTNR1A gene has both additive and dominant effects on total number of born piglets (TNB) and number of piglets born alive (NBA). Additive effects were significant in summer (TNB, P < 0.01; NBA, P < 0.001), whereas dominant effects reached significance both in fall (TNB, P < 0.01; NBA, P < 0.05) and in winter (TNB, P < 0.001; NBA, P < 0.05). The seasonal variation observed for MTNR1A additive and dominant effects might be produced by the influence of photoperiod on the pattern and duration of melatonin secretion. These results illustrate that the complex interaction between genotype and environment can be an important source of phenotypic variation of reproductive traits.     

Corpus luteum size and plasma progesterone concentration in cows

It is often assumed that a larger corpus luteum will produce more progesterone and generate higher circulating plasma concentrations. The aim of the study was to determine whether the size of the corpus luteum does actually determine circulating plasma progesterone concentrations. Data were collated from a number of studies on various aspects of luteal function in non-lactating dairy cows to allow comparisons to be made between corpus luteum weight and plasma progesterone concentration across the luteal phase. In these studies oestrous cycles had been synchronised and animals slaughtered on day 5, day 8 or day 16 following oestrus. Both corpus luteum weight and plasma progesterone concentration increased between day 5 and day 8. Plasma progesterone concentration but not luteal weight also increased between day 8 and day 16. On day 5 there was a strong relationship between corpus luteum weight and plasma progesterone (R² = 0.64; P < 0.001). However, no such relationship was present on day 8 or day 16. These results indicate that while during the early stage of corpus luteum development a relationship between size and progesterone is present, by day 8 of the cycle, the size of the corpus luteum is no longer of importance in determining circulating progesterone concentrations.     

Endocrine changes during pregnancy, parturition and post-partum in guanacos (Lama guanicoe)

Plasma concentrations of progesterone (P4), estradiol-17β (E2), estrone (E1) and estrone sulfate (E1S) were measured during gestation in eight guanacos kept in captivity. Gestational length was 346.1 ± 9.8 days. P4 plasma concentrations increased after ovulation and remained elevated until parturition. However, during the last 4 weeks of gestation, a gradual decrease from 4.17 × 1.17^±1 nmol/L to 2.02 × 1.95^±1 nmol/L on day 5 before parturition was observed, followed by a more abrupt final decline to baseline concentrations which were reached on the day after parturition. Mean E2 plasma concentrations started to increase during the eighth month of gestation, and were significantly elevated up to maximum concentrations of 484.7 × 1.21^±1 pmol/L during the last 2 months of pregnancy. Concentrations returned to baseline during the last 2 days of gestation. An increase of E1S concentrations (p < 0.01) was observed in the eleventh month of gestation. Mean E1S concentrations remained rather constant during the last 3 weeks of gestation between 4 to 8 nmol/L until parturition, when a steep precipitous decline was observed. E1 concentrations were slightly elevated during the last 4 weeks of gestation, however, maximum concentrations did not exceed 1.5 nmol/L. The results show distinct species specific features of gestational steroid hormone profiles in the guanaco in comparison to domestic South American camelids, such as a more pronounced gradual prepartal decrease of P4 concentrations prior to the final decline to baseline, and clearly lesser E1S concentrations during the last 4 weeks of gestation, which lack a continuous prepartal increase.     

Allelic switching of the imprinted IGF2R gene in cloned bovine fetuses and calves

Cloned animals often suffer from loss of development to term and abnormalities, typically classified under the umbrella term of Large Offspring Syndrome (LOS). Cattle are an interesting species to study because of the relatively greater success rate of nuclear transfer in this species compared with all species cloned to date. The imprinted insulin-like growth factor receptor (IGF2R; mannose-6-phosphate) gene was chosen to investigate aspects of fetal growth and development in cloned cattle in the present study. IGF2R gene expression patterns in identical genetic clones of several age groups were assessed in day 25, day 45, and day 75 fetuses as well as spontaneously aborted fetuses, calves that died shortly after birth and healthy cloned calves using single stranded conformational polymorphism gel electrophoresis. A variable pattern of IGF2R allelic expression in major organs such as the brain, cotyledon, heart, liver, lung, spleen, kidney and intercotyledon was observed using a G/A transition in the 3’UTR of IGF2R. IGF2R gene expression was also assessed by real time RT-PCR and found to be highly variable among the clone groups. Proper IGF2R gene expression is necessary for survival to term, but is most likely not a cause of early fetal lethality or an indicator of postnatal fitness. Contrary to previous reports of the transmission of imprinting patterns from somatic donor cells to cloned animals within organs in the same cloned animal the paternal allele of IGF2R can be imprinted in one tissue while the maternal allele is imprinted in another tissue. This observation has never been reported in any species in which imprinting has been studied.