A cDNA library from the antenna of Monochamus alternatus Hope and binding properties of odorant‐binding proteins

Chemoreception is a key feature in selection of host plants by insects. We performed a preliminary functional characterization of olfactory proteins isolated from an antennal cDNA library of Monochamus alternatus. We identified four olfactory genes, including two encoding putative classic odorant‐binding proteins (OBPs) and two encoding minus‐C OBPs. We expressed two of the four OBPs, MaltOBP3 and MaltOBP5, in a bacterial system and assessed their ligand specificity by measuring the competitive binding of fluorescent probe, N‐phenyl‐1‐naph‐thylamine, in the presence of 17 volatile beetle‐ or host‐plant‐related ligands. The results indicated that although MaltOBP3 and MaltOBP5 bound a distinctly different group of competitors, both had relatively high binding affinities (K_i < 20 μm ) for certain compounds. The differences in their binding affinities towards host‐plant ligands suggest the roles of MaltOBP3 and MaltOBP5 in host‐plant selection.     

β‐Ionone as putative semiochemical suggested by ligand binding on an odorant‐binding protein of Hylamorpha elegans and electroantennographic recordings

Currently, odorant‐binding proteins (OBPs) are considered the first filter for olfactory information for insects and constitute an interesting target for pest control. Thus, an OBP (HeleOBP) from the scarab beetle Hylamorpha elegans (Burmeister) was identified, and ligand‐binding assays based on fluorescence and in silico approaches were performed, followed by a simulated binding assay. Fluorescence binding assays showed slight binding for most of the ligands tested, including host‐plant volatiles. A high binding affinity was obtained for β‐ionone, a scarab beetle‐related compound. However, the binding of its analogue α‐ionone was weaker, although it is still considered good. On the other hand, through a three‐dimensional model of HeleOBP constructed by homology, molecular docking was carried out with 29 related ligands to the beetle. Results expressed as free binding energy and fit quality (FQ) indicated strong interactions of sesquiterpenes and terpenoids (α‐ and β‐ionone) with HeleOBP as well as some aromatic compounds. Residues such as His102, Tyr105 and Tyr113 seemed to participate in the interactions previously mentioned. Both in silico scores supported the experimental affinity for the strongest ligands. Therefore, the activity of α‐ionone, β‐ionone and 2‐phenyl acetaldehyde at antennal level was studied using electroantenography (EAG). Results showed that the three ligands are electrophysiologically active. However, an aliquot of β‐ionone (represented by 3.0 ng) elicited stronger EAG responses in antennae of males than of females. Finally, the role of these ligands as potential semiochemicals for H. elegans is discussed.     

Assessing cultivar resistance to Sitodiplosis mosellana (Géhin) (Diptera: Cecidomyiidae) using a phenotyping method under semi‐field conditions

The orange wheat blossom midge, Sitodiplosis mosellana (Géhin), can significantly reduce wheat yield. Growing resistant wheat cultivars is an effective way of managing this pest. The assessment of cultivar resistance in field trials is difficult because of unequal pressure of S. mosellana caused by differences in cultivar heading dates relative to the flight period of S. mosellana adult females and huge variations of egg laying conditions from 1 day to another. To overcome these hurdles and to expose all cultivars homogeneously to the pest, an assessment method of cultivar resistance was developed under semi‐field conditions. In 2015, the resistance of 64 winter wheat cultivars to S. mosellana was assessed. Few or no larvae developed in the ears of resistant cultivars, but in susceptible cultivars, large numbers of larvae developed. Seventeen cultivars proved to be resistant, whereas 47 were susceptible. The identification of new resistant cultivars offers more opportunities to manage S. mosellana. The phenotyping method is easy, cheap, efficient and reliable. It can be used to guide the breeding of new resistant wheat cultivars. Using specific midge populations, this method could also be used in research on new resistance mechanisms in winter wheat or in other cereal species.     

Molecular cloning,expression profile,odorant affinity,and stability of two odorant‐binding proteins in Macrocentrus cingulum Brischke (Hymenoptera: Braconidae)

The polyembryonic endoparasitoid wasp Macrocentrus cingulum Brischke (Hymenoptera: Braconidae) is deployed successfully as a biocontrol agent for corn pest insects from the Lepidopteran genus Ostrinia in Europe and throughout Asia, including Japan, Korea, and China. The odorants are recognized, bound, and solubilized by odorant‐binding protein (OBP) in the initial biochemical recognition steps in olfaction that transport them across the sensillum lymph to initiate behavioral response. In the present study, we examine the odorant‐binding effects on thermal stability of McinOBP2, McinOBP3, and their mutant form that lacks the third disulfide bonds. Real‐time PCR experiments indicate that these two are expressed mainly in adult antennae, with expression levels differing by sex. Odorant‐binding affinities of aldehydes, terpenoids, and aliphatic alcohols were measured with circular dichroism spectroscopy based on changes in the thermal stability of the proteins upon their affinities to odorants. The obtained results reveal higher affinity of trans‐caryophelle, farnesene, and cis‐3‐Hexen‐1‐ol exhibits to both wild and mutant McinOBP2 and McinOBP3. Although conformational flexibility of the mutants and shape of binding cavity make differences in odorant affinity between the wild‐type and mutant, it suggested that lacking the third disulfide bond in mutant proteins may have chance to incorrect folded structures that reduced the affinity to these odorants. In addition, CD spectra clearly indicate proteins enriched with α‐helical content.     

Electroantennogram response and attraction of Anastrepha suspensa to volatiles of various sugar sources and aged sugar solutions

With the aim of finding new, sugar‐based volatile attractants for economically important tephritid fruit fly species, we used electroantennography (EAG) to quantitate olfactory responses of female Caribbean fruit fly, Anastrepha suspensa (Loew) (Diptera: Tephritidae), to volatiles of six sugar sources (refined white and brown cane sugar, coconut sugar, date sugar, date jaggery, and cane panela). Laboratory‐strain and wild flies, both sexually immature and mature, were tested for EAG responses to the volatiles of dry crystallized sugar sources and 10% (wt/vol) aqueous solutions that had aged in the laboratory for 0–7 days. In general, wild flies exhibited higher EAG responses than laboratory flies, and immature females responded more strongly than mature females. With the exception of date jaggery and cane panela, volatiles of dry sugar sources and 0‐ and 1‐day‐old solutions elicited lower EAG responses than any of the aged solutions. Most solution volatiles elicited the strongest EAG response after 2 days of aging. Of the treatments evaluated, volatiles of the 5‐day‐old date jaggery solution elicited the highest‐amplitude EAG responses (39%) in A. suspensa females. On the basis of the latter, we tested the attraction response of mature and immature females to date‐jaggery solutions aged over 2 and 4 days in two‐choice flight tunnel bioassays. With both mature and immature females, the 2‐day‐old solution was more attractive than the 4‐day‐old jaggery solution, but significantly more mature females (70% of captures) were attracted to 2‐day‐old jaggery solution. We discuss our results with respect to the improvement of fruit fly lures and attractants by incorporating elements from aged date‐jaggery sugar.     

Antennal sensitivity to female sex pheromone compounds of Spodoptera frugiperda males (Lepidoptera: Noctuidae) and associated field behaviour

The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is one of the most important insect pests of corn, sorghum, rice, and grasses. The sex pheromone produced by S. frugiperda is composed of a mixture of esters. In this study, I determined the antennal responses of FAW males from 22 populations in Mexico to the components of the sex pheromone of this species: (Z)-7-dodecenyl acetate (Z7-12:OAc), (Z)-9-tetradecenyl acetate (Z9-14:OAc), and (Z)-11-hexadecenyl acetate (Z11-16:OAc) at four doses (0.01, 0.1, 1, and 10 μg). In addition, I evaluated the behavioural activity of single compounds in the field. The largest antennal responses of FAW males from all the sampled regions were elicited by Z9-14:OAc (all doses evaluated) and Z7-12:OAc (0.01, 0.1, and 1 μg). The male antennal responses evoked by Z11-16:OAc were smaller but similar to responses to Z7-12:OAc at lower doses (0.01 and 0.1 μg). In the field test, Z9-14:OAc produced significantly higher captures of S. frugiperda males compared to Z7-12:AOc and Z11-16:OAc. Antennal responses were related to captures obtained in the field, supported by the positive correlation of these variables [electroantennography (EAG) response and behaviour], with the exception of Z7-12:OAc, which elicited relatively large EAG responses but did not produce high captures of S. frugiperda. These results are of importance to agriculture, enabling the implementation of better methods of monitoring and controlling this pest, and support the suggestion that a mixture of Z9-14:OAc and Z7-12:OAc could represent an effective attractant for all evaluated populations of S. frugiperda in Mexico.     

Volatiles released from foliar extract of host plant enhance landing rates of gravid Polygonia c‐album females,but do not stimulate oviposition

The role of olfactory cues for host search is much less investigated in day‐active butterflies than in their relatives, the nocturnal moths. The goal of this study was to investigate whether host‐plant volatiles from foliar extracts of hop, Humulus lupulus L. (Cannabaceae), evoke electroantennographic (EAG) responses, increase landing rates, and stimulate egg‐laying behavior of gravid Polygonia c‐album L. (Lepidoptera: Nymphalidae) females. Eighty‐nine volatile compounds were detected in a non‐concentrated methanol extract of hop by gas chromatography–mass spectrometry, 11 of which elicited an EAG response. Concentration of the crude extract significantly reduced landing rates on artificial leaves treated with the sample due to loss of volatile compounds, but after landing the oviposition response of gravid females was not affected. A mixture of eight commercially available EAG‐active volatiles increased the landing rate of gravid females to their source but did not act as oviposition stimulants. Dividing the volatile compounds into two groups – consisting of (1) hexanal, (E)‐2‐hexenal, octanal, nonanal, and decanal, and (2) sulcatone, humulene, and benzyl alcohol – obliterated effectiveness, revealing synergism between compounds. Although volatiles did not stimulate oviposition, they significantly contributed to the distribution of eggs by increasing the landing rates on treated artificial leaves.     

Characterization and Distribution of a Cloned Rat μ-Opioid Receptor

Abstract: We have cloned and expressed a rat brain cDNA, TS11, that encodes a μ-opioid receptor based on pharmacological, physiological, and anatomical criteria. Membranes were prepared from COS-7 cells transiently expressing TS11 bound [³H]diprenorphine with high affinity (K_D = 0.23 ± 0.04 nM). The rank order potency of drugs competing with [³H]diprenorphine was as follows: levorphanol (K_i = 0.6 ± 0.2 nM) ≈β-endorphin (K_i = 0.7 ± 0.5 nM) ≈ morphine (K_i = 0.8 ± 0.5 nM) ≈ [d -Ala², N-Me-Phe⁴,Gly-ol⁵]-enkephalin (DAMGO; K_i = 1.6 ± 0.5 nM) ? U50,488 (K_i = 910 ± 0.78 nM) > [d -Pen^2,5]-enkephalin (K_i = 3,170 ± 98 nM) > dextrorphan (K_i = 4,100 ± 68 nM). The rank order potencies of these ligands, the stereospecificity of levorphanol, and morphine's subnanomolar K_i are consistent with a μ-opioid binding site. Two additional experiments provided evidence that this opioid-binding site is functionally coupled to G proteins: (a) In COS-7 cells 50 µM 5′-guanylylimidodiphosphate shifted a fraction of receptors with high affinity for DAMGO (IC₅₀ = 3.4 ± 0.5 nM) to a lower-affinity state (IC₅₀ = 89.0 ± 19.0 nM), and (b) exposure of Chinese hamster ovary cells stably expressing the cloned μ-opioid receptor to DAMGO resulted in a dose-dependent, naloxone-sensitive inhibition of forskolin-stimulated cyclic AMP production. The distribution of mRNA corresponding to the μ-opioid receptor encoded by TS11 was determined by in situ hybridization to brain sections prepared from adult female rats. The highest levels of μ-receptor mRNA were detected in the thalamus, medial habenula, and the caudate putamen; however, significant hybridization was also observed in many other brain regions, including the hypothalamus.     

Electroantennogram responses of aphid nymphs to plant volatiles

Abstract. Electroantennogram (EAG) responses of immature aphids were investigated for 30 plant volatile compounds in third‐ and fourth‐stadium nymphs of the black bean aphid, Aphis fabae. The nymphs were destined to develop into adult alate (winged) virginoparae. The EAG response profiles were similar to those previously reported in the adults. Among the compounds tested, hexanonitrile elicited the largest EAG responses in both nymphal stadia, corresponding to previously reported results with adults. Six‐carbon aliphatic compounds showed relatively higher EAG activities in the nymphs but, in contrast, (E)‐2‐hexenal, benzaldehyde, α‐pinene and β‐pinene, and citronellal elicited relatively smaller EAG responses in nymphs than adults. Although overall EAG response profiles were similar between the third and the fourth stadia for the majority of the volatiles, four aldehyde compounds, hexanal (E)‐2‐heptenal, 2‐hydroxybenzaldehyde and citronellal, showed relatively higher EAG activities in the third than in the fourth stadium. The present study indicates that aphid nymphs possess a functional olfactory receptor system before the antennae are fully developed morphologically and physiologically.     

青杨脊虎天牛对植物源挥发物的EAG和行为反应

测定了青杨脊虎天牛Xylotrechus rusticus （L.）雌、雄成虫对其寄主杨树中的水杨醛（0.95 μmol/μL）和非寄主植物中0.3 μmol/μL的叶绿醇、0.4 μmol/μL的水芹烯和0.6 μmol/μL的 R 型α-蒎烯、S 型α-蒎烯、S型β-蒎烯、3-蒈烯、罗勒烯、香草烯和松节油等10种植物挥发性气味物质的触角电位（EAG）反应。结果表明,与对照相比,这10种植物挥发物多能引起成虫明显的EAG反应( P<0.05,P<0.01 ),其中雌虫对松节油、水杨醛、R 型α-蒎烯和 S 型α-蒎烯的EAG反应较强; 雄虫对 R 型α-蒎烯的EAG反应最强,松节油次之。根据雌虫对这10种挥发物EAG反应的强弱,进一步测定了雌虫对0.00006、0.0006、0.006、0.06、0.6、0.12 μmol/μL的松节油、R 型α-蒎烯、S 型α-蒎烯以及0.000095、0.00095、0.0095、0.095、0.95、0.19 μmol/μL的水杨醛的EAG和行为反应。结果表明,雌虫对松节油、水杨醛和 R 型α-蒎烯的EAG反应随气味物质浓度的增加而增加,水杨醛浓度增加到0.95 μmol/μL、松节油和 R 型α-蒎烯浓度增加到0.6 μmol/μL以后,EAG反应值趋于平稳;对 S 型α-蒎烯的反应随浓度的增加而呈线性增加。水杨醛浓度低于0.095时,对雌虫没有明显的定向作用( P>0.05 ),高于此浓度时表现为驱避作用( P<0.05 ); 松节油在浓度低于或等于0.6 μmol/μL时对雌虫表现为驱避作用,浓度为0.6时驱避效果最佳（ P<0.01 ）。雌虫对 R 型α-蒎烯和 S 型α-蒎烯没有明显的定向行为反应。     

Stereoselective binding of isradipine to human plasma proteins

Isradipine (PN 200–110) is a highly potent calcium entry blocker with an asymmetrically substituted dihydropyridine ring (methyl- and isopropylester, respectively). The binding of the (+)-(S)-isradipine and (?)-(R)-isradipine to isolated human serum albumin (HSA, 30 μmol/l) and α₁-acid glycoprotein (AAG, 10 μmol/l) has been studied in vitro over a wide range of isradipine concentrations (0.06–20 μmol/l) using high-performance liquid chromatography (HPLC). HPLC experiments revealed that both isradipine enantiomers were bound to one class of high-affinity binding sites on the AAG molecule (n_(S) = 0.83 ± 0.05, K_a(S) = (1.33 ± 0.25) × 10⁶ 1/mol, n_(R) = 0.85 ± 0.07, K_a(R) = (1.17 ± 0.44) × 10⁷ l/mol). The (R)-enantiomer also exhibited an interaction with the secondary low-affinity binding sites (n′K′_{a (R)} = (2.66 ± 0.65) × 10⁴ l/mol). In contrast, the pharmacologically more potent (+)-(S)-enantiomer was more strongly bound to HSA than its optical antipode (n_(S) = 1.07 ± 0.07, K_a(S) = (1.76 ± 0.26) × 10⁵ l/mol, nK_a(R) = (3.62 ± 0.06) × 10⁴ l/mol). In general, the resulting binding characteristics of individual isradipine enantiomers showed stereoselectivity, but this was opposite for the two most important plasma binding proteins. The process of accumulation of isradipine by human platelets in the therapeutically relevant range (10–80 ng/ml) at 37°C was devoid of stereoselectivity. © 1995 Wiley-Liss, Inc.     

Comparison of the olfactory sensitivity of two sympatric steppe grasshopper species (Orthoptera: Acrididae) to plant volatile compounds

Electroantennogram (EAG) responses of male and female Oedipodinae grasshoppers, Oedaleus decorus asiaticus B.-Bienko and Angaracris barabensis Pall to 37 plant volatile compounds were recorded. The two species have sympatric distribution and synchronous seasonal activity in Inner Mongolia Grasslands. They have different host plant preference with Oedaleus decorus asiaticus graminivorous and A. barabensis forbivorous. The EAG response profiles to 37 compounds of the two species and their both sexes were similar. Most of the green leaf volatiles elicited higher EAG responses in both species and sexes than terpenic compounds and some aromatic compounds. Strong EAG responses were elicited by 6-carbon alcohols (1-hexanol, Z-hexen-2-ol-1, E-2-hexen-1-ol, E-hexen-3-ol-1), aldehyde (E-2-hexen-1-al), ester (Z-hexen-3-yl acetate), and 7-carbon alcohols (1-heptanol) in both species and sexes. Monoterpenes with functional groups of alcohols and aldehydes were more stimulating than other monoterpenes tested. The sesquiterpenes tested elicited relatively low responses. Benzaldehyde elicited the highest responses for both species among aromatic components. Oedaleus decorus asiaticus showed higher EAG responses than A. barabensis to the green leaf volatiles, 1-decanol, 1-nonanol, 1-pentanol, hexanal, Z-hexen-3-yl acetate and to the sesquiterpenes (-)-trans-caryophyllene. Males have higher responses than females in Oedaleus decorus asiaticus. No sexual difference was observed in A. barabensis. Dose-dependent responses to six selected chemicals were observed from females. In both species, all the chemicals tested elicit EAG responses at concentration between 10^-3 mol/L and 10^-2 mol/L, except that the responses of A. barabensis to terpineol had a threshold concentration of 10-2 mol/L benzaldehyde and 1-hexanol had the highest slopes in dose curves, while 1-octen-3-ol showed the smallest slope in responses to the six chemicals tested. Comparative studies on the responses of two antennal sections and the whole antenna to six selected chemicals were carried out using females of both species. A significant EAG response was only recorded from the distal part of the antenna and not from the proximal seven segments.     

The strengths and limitations of using biolayer interferometry to monitor equilibrium titrations of biomolecules

Every method used to quantify biomolecular interactions has its own strengths and limitations. To quantify protein‐DNA binding affinities, nitrocellulose filter binding assays with ³²P‐labeled DNA quantify K_d values from 10^?12 to 10^?8 M but have several technical limitations. Here, we considered the suitability of biolayer interferometry (BLI), which monitors association and dissociation of a soluble macromolecule to an immobilized species; the ratio k_off/k_on determines K_d. However, for lactose repressor protein (LacI) and an engineered repressor protein (“LLhF”) binding immobilized DNA, complicated kinetic curves precluded this analysis. Thus, we determined whether the amplitude of the BLI signal at equilibrium related linearly to the fraction of protein bound to DNA. A key question was the effective concentration of immobilized DNA. Equilibrium titration experiments with DNA concentrations below K_d (equilibrium binding regime) must be analyzed differently than those with DNA near or above K_d (stoichiometric binding regime). For ForteBio streptavidin tips, the most frequent effective DNA concentration was ~2 × 10^?9 M. Although variation occurred among different lots of sensor tips, binding events with K_d ≥ 10^?8 M should reliably be in the equilibrium binding regime. We also observed effects from multi‐valent interactions: Tetrameric LacI bound two immobilized DNAs whereas dimeric LLhF did not. We next used BLI to quantify the amount of inducer sugars required to allosterically diminish protein‐DNA binding and to assess the affinity of fructose‐1‐kinase for the DNA‐LLhF complex. Overall, when experimental design corresponded with appropriate data interpretation, BLI was convenient and reliable for monitoring equilibrium titrations and thereby quantifying a variety of binding interactions.     

Floral volatiles with colour cues from two cucurbitaceous plants causing attraction of Aulacophora foveicollis

Aulacophora foveicollis Lucas (Coleoptera: Chrysomelidae) is an important phytophagous pest of two cucurbitaceous plants, Momordica cochinchinensis Spreng and Solena amplexicaulis (Lam.) Gandhi. The volatile organic compound profiles from flowers of M. cochinchinensis and S. amplexicaulis were identified and quantified by gas chromatography‐mass spectrometry (GC‐MS) and GC‐flame ionization detector (FID) analyses. Twenty nine and 28 compounds were identified in volatiles of M. cochinchinensis and S. amplexicaulis flowers, respectively. Methyl jasmonate and 3‐octanol were the predominant volatiles of M. cochinchinensis flowers, whereas 1‐octadecanol and 1‐hexanol were most found in the headspace of S. amplexicaulis flowers. Aulacophora foveicollis were more attracted by the flower volatiles of M. cochinchinensis than by those of S. amplexicaulis in a glass Y‐tube olfactometer. A mixture of 1‐heptanol, linalool oxide, 1‐octanol, and nonanal in the proportions present in the headspace of both flower types elicited attraction in the insect. From 25 cm distance, A. foveicollis displayed a preference for artificial flowers of 6.5 cm diameter of S. amplexicaulis flower colour (white) over M. cochinchinensis flower colour (white‐yellow). Finally, a synthetic blend (0.43 μg 1‐heptanol + 1.44 μg linalool oxide + 0.14 μg 1‐octanol + 1.77 μg nonanal dissolved in 25 μl methylene chloride) attracted more beetles when applied in a white artificial flower than when applied in a white‐yellow artificial flower from 40 cm distance. This finding may be helpful in the development of traps for pest management strategies.     

Field efficacy of insect pathogen,botanical, and jasmonic acid for the management of wheat midge Sitodiplosis mosellana and the impact on adult parasitoid Macroglenes penetrans populations in spring wheat

The wheat midge, Sitodiplosis mosellana, is a serious pest of wheat worldwide. In North America, management of S. mosellana in spring wheat relies on the timely application of pesticides, based on midge adults levels caught in pheromone traps or seen via field scouting during wheat heading. In this context, biopesticides can be an effective alternative to pesticides for controlling S. mosellana within an Integrated Pest Management program. A field study using insect pathogenic fungus Beauveria bassiana GHA, nematode Steinernema Jeltiae with Barricade polymer gel 1%, pyrethrin, combined formulations of B. bassiana GHA and pyrethrin, Jasmonic acid (JA) and chlorpyrifos (chemical check) was performed to determine to which extent they affect midge larval populations, kernel damage levels, grain yield, and quality, and the impacts on adult parasitoid Macroglenes penetrans populations. The results indicated that biopesticides JA and S. Jeltiae were the most effective in reducing larval populations and kernel damage levels, and produced a higher spring wheat yield when compared to the water control at both study locations (East Valier and North Valier, Montana, USA). Increased test weight in wheat had been recorded with two previous biopesticides at East Valier but not for North Valier, when compared over water control. These results were comparable in efficacy to the chlorpyrifos. This study also suggested that B. bassiana and pyrethrin may work synergistically, as exemplified by lower total larval populations and kernel damage levels when applied together. This study did not demonstrate the effect of any treatments on M. penetrans populations.     

Electroantennographic responses of red flour beetle Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) to volatile organic compounds

The red flour beetle Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is the most encountered and destructive stored product insect pest of cereal grains and seeds. Although this beetle has been used as a model organism for many decades, there is no systematic knowledge about antennal detection of host and non‐host volatiles. Electroantennogram responses to 94 selected volatile organic compounds including alkanes, alkenes, alcohols, aldehydes, ketones, carboxylic acids, esters, terpenoids and aromatic compounds were recorded from both sexes of T. castaneum. Overall, female and male T. castaneum exhibited similar electroantennography (EAG) responses. Compounds eliciting the strongest EAG responses within compound groups of chemical similarity were undecane, 1‐hexen‐3‐ol, octanal, 2‐heptanone, hexanoic acid and ethyl hexanoate. Comparison of vapour pressure and EAG amplitudes within homologous series of compounds revealed responses to undecane, octadecane, octanal, nonanal, 2‐heptanone, hexanoic acid and octanoic acid as outstanding. Given that systematic EAG screenings have not been conducted before, these are the best candidates for evaluation in future behavioural studies to unravel their potential for application in integrated pest management strategies of T. castaneum.     

Characterization of [3H]LS‐3‐134, a novel arylamide phenylpiperazine D3 dopamine receptor selective radioligand

LS‐3‐134 is a substituted N‐phenylpiperazine derivative that has been reported to exhibit: (i) high‐affinity binding (K_i value 0.2 nM) at human D3 dopamine receptors, (ii) > 100‐fold D3 versus D2 dopamine receptor subtype binding selectivity, and (iii) low‐affinity binding (K_i > 5000 nM) at sigma 1 and sigma 2 receptors. Based upon a forskolin‐dependent activation of the adenylyl cyclase inhibition assay, LS‐3‐134 is a weak partial agonist at both D2 and D3 dopamine receptor subtypes (29% and 35% of full agonist activity, respectively). In this study, [³H]‐labeled LS‐3‐134 was prepared and evaluated to further characterize its use as a D3 dopamine receptor selective radioligand. Kinetic and equilibrium radioligand binding studies were performed. This radioligand rapidly reaches equilibrium (10–15 min at 37°C) and binds with high affinity to both human (K_d = 0.06 ± 0.01 nM) and rat (K_d = 0.2 ± 0.02 nM) D3 receptors expressed in HEK293 cells. Direct and competitive radioligand binding studies using rat caudate and nucleus accumbens tissue indicate that [³H]LS‐3‐134 selectively binds a homogeneous population of binding sites with a dopamine D3 receptor pharmacological profile. Based upon these studies, we propose that [³H]LS‐3‐134 represents a novel D3 dopamine receptor selective radioligand that can be used for studying the expression and regulation of the D3 dopamine receptor subtype.