On the sense of taste in two Malagasy Primates (Microcebus murinus and Eulemur mongoz)

The relationship between phylogeny and taste is of growing interest.In this study we present recordings from the chorda tympaniproper (CT) nerve of two lemuriforme primates, the lesser mouselemur (Microcebus murinus) and the mongoose lemur (Eulemur mongoz),to an array of taste stimuli which included the sweeteners acesulfame-K,alitame, aspartame, D-glucose, dulcin, monellin, neohesperidindihydrochalcone (NHDHC), saccharin, sodium superaspartame, stevioside,sucralose (TGS), sucrose, suosan, thaumatin and xylitol, aswell as the non–sweet stimuli NaC1, citric acid, tanninand quinine hydrochloride. In M.murinus the effects of the tastemodifiers gymnemic acid and miraculin on the CT response wererecorded. Conditioned taste aversion (CTA) experiments in M.murinusand two-bottle preference (TBP) tests in E.mongoz were alsoconducted. We found that all of the above tastants except thaumatinelicited a CT response in both species. The CTA technique showedthat M.murinus generalized from sucrose to monellin but notto thaumatin. The intake of aspartame, ranging in concentrationfrom 0.1 to 30 mM was measured in E.mongoz with TBP tests. Atno concentration did we see a preference, but there was a significantrejection of 10 and 30 mM aspartame (P     

Gustatory responses in primates to the sweetener aspartame and their phylogenetic implications

Two-bottle preference tests have been applied to 70 (sub-) speciesof the order of Primates and, for comparison, to the tree shrew(Tupaia belangeri) to determine their responses to aspartame(APM), the first known sweet-tasting dipeptide which has toman a sweetness potency of about 200 times that of sucrose.It was found that only the Cercopithecidae, the Hylobatidaeand the Pongidae respond like man to this dipeptide and preferit to water. The other primates tested to date, show no responseto this sweetener. From a phylogenetic point of view, we notethat APM shows species specificity similar to thaumatin. Thus,a clear dichotomy exists within the order of Primates with respectto both thaumatin and APM. The results here illustrate oncemore the gustatory diversity among primates and are a compellingargument for the existence of different sweet taste receptorsor recognition sites in primate species.     

ON THE GUSTATORY EFFECTS OF MONELLIN AND THAUMATIN IN DOG, HAMSTER, PIG AND RABBIT

The electrical activity in the chorda tympani proper nerve ofdog, hamster, pig and rabbit was recorded during stimulationof the tongue with the sweet proteins, monellin and thaumatin,and stimuli representing the four taste qualities. It was observedthat these proteins, which to man taste extremely sweet andin the monkey elicit a significant neural response, caused,except for monellin in the dog, no significant change in theneural activity. On the basis of these results it is suggestedthat different types of ‘sweet’ receptor sites existin mammals.     

Gustatory responses in three prosimian and two simian primate species (Tupaia glis, Nycticebus coucang, Galago senegalensis, Callithrix jacchus jacchus and Saguinus midas niger) to six sweeteners and miraculin and their phylogenetic implications

The intake of six sweeteners was recorded together with theireffects on the impulse activity of the chorda tympani propernerve during their application to the tongue. The sweetenerswere: acetosulfam, aspartame, D-tryptophan, glycine, xylitoland thaumatin. They were used at human equi-sweet concentrations.In all species, D-tryptophan was strongly preferred and gavea significant response, while aspartame and thaumatin gave neithera significant behavioral nor a significant neural response.Acetosulfam, glycine and xylitol elicited neural responses,but their behavioral effects differed from a rejection in somespecies to a preference in others. Miraculin, which has a sweetnessinducing effect in man, showed this effect only in the platyrrhineanspecies and not in the prosimian.     

Study of the effect of gymnemic acid on taste in hamster

Behavioral and electrophysiological experiments with 10 sweetenershave been made to test if gymnemic acid (GA) is able to blockthe response to sweet stimuli in single taste fibres of thechorda tympani proper nerve in hamsters. The hamster was chosenbecause earlier studies show that it is more sensitive to GAthan any other non-human species. Since GA has been shown toaffect the sweetness of many different substances, its effectswere studied on an array of sweeteners. To avoid, however, theinclusion of sweeteners unpalatable to the hamster, the hamsters'liking of this array was tested with two-bottle preference techniquefor –24 h. It was found that acesulfam-K, fructose, glucose,sucrose and xylitol were strongly liked, while the animals showedno preference for aspartame, D-tryptophane, sodium cyclamate,sodium saccharin and thaumatin over water. The summated nerveresponse of these stimuli was then recorded. It was found thatneither thaumatin nor aspartame elicited a response, while theother stimuli gave a good response. Finally, the sweetenerswhich were both preferred in the two-bottle tests and gave anerve response were used as taste stimuli in single fibre experimentstogether with sodium chloride, quinine hydrochloride, citricacid and saccharin. The single fibre recordings were made beforeand after application of 5 mg GA for 3 min on the tongue. Itwas found that GA did not cause any dramatic decrease or disappearanceof the responses to either the sweet or the non-sweet substances.The responses to the sweeteners, however, were more depressedthan those to the non-sweet stimuli.     

Catecholaminergic contributions to the neuronal machinery of the olfactory bulb: aftoradiographic, immunohistochemical and evoked feild potential studies

aftographic exeperiments on the localization of radiolabelednoradrenaline, dopamine and dopa, as well as immunohistochemicalstudies on hydroxylase-like activity, are summarized and comparedin both rat and turtle olfactory bulbs. Evoked field potentialstudies on effects of dopamine are also discussed. The histochemicalstudies suggest that dopaminergic periglomerular neurons arethe most significant cellular component of the catecholaminergicsystem in the olfactory bulb of both species. Scattered fluorescentcell group was also present in the internal plexiform layerand superficial granule cell layer of the turtle olfactory bulb.Other fibres, not related to intrinsic bulbar neuronal cellbodies, were also labeled, mostly in the granule cell layerbut also in the external plexiform layer. These might belongto a centrifugal catecholaminergic system from brain stem neurons.In the in vitro turtle olfactory bulb, dopamine and apomorphinedepressed the amplitude of field potentials evoked by a singlevolley in the olfactory nerve or lateral olfactory tract, andreduced the depression and latency of reponses when paired volleywere delivered. It is suggested that catecholaminergic systemsplay a key role in modulating mitral cell activity through actionsin both superficial (glomerular) and deep (granule) layers.This may involve direct actions, or other, non-catecholaminergicinterneurons.     

The Effect of Hypoxia and Sulphide on Culture-Grown Wetland and Non-Wetland Plants: II. METABOLIC AND PHYSIOLOGICAL CHANGES

Pearson, J. and Havill, D. C. 1988. The effect of hypoxia andsulphide on culture-grown wetland and non-wetland plants. II.Metabolic and physiological changes.—J. exp. Bot. 39:431–439. Two non-wetland (Agropyron pungens, Hordeum vulgare) and fivewetland species (Oryza sativa, Aster tripolium, three Salicorniaspp.) were grown in aerated, unaerated and sulphide-treatedculture solution. Changes in the activity of alcohol dehydrogenase(ADH) and cytochrome oxidase (COase) in the roots were measured.In the non-wetland species, treatment with hypoxia or sulphideincreased ADH activity by 900–1 800%, whereas COase activitydecreased by 80–92% of the aerated control. In the wetlandspecies, except S. europaea which was not affected, hypoxiaincreased ADH activity by 350–550%, while COase activitywas little affected. Generally, when treated with sulphide theactivity of ADH increased to about 750% in most of the wetlandspecies, but increases as low as 175% (S. europaea) and as highas 1400% (S.fragilis) were recorded. The effect of sulphideon the COase activity in the wetland plants was not as markedas in the non-wetland plants. The Salicornia spp. were the leastaffected by the sulphide treatment and they also had intrinsicallyhigher levels of COase activity than the other species sampled.Of the wetland plants the Salicornia species had the lowestvalue for root aerenchyma, 3–6%. Therefore, there wasno correlation between the possession of aerenchyma and thephysiological changes measured. Measurement of malate, lactateand ethanol in roots of the first four species listed abovegave no evidence for alternative anaerobic fermentation pathways.While in the flood-intolerant species, high ADH activities werenot able to maintain the energy charge. It is suggested thatmaintenance of relatively high COase activity in wetland plantsmay help to ‘scavenge’ any available oxygen withinroots and thus help reduce energy loss. Key words: Cytochrome oxidase, alcohol dehydrogenase, metabolic adaptation     

Anterograde and retrograde axonal transport of 6-phosphofructokinase activity in the rat sciatic nerve

Optimal conditions for the measurement of phosphofructokinase activity in segments of rat sciatic nerve were established. It was found that maximal activity was obtained when the Triton X-100 concentration of the extraction buffer was 1% (v:v). Nerve segments could be stored at −80°C for at least 1 week without measurable loss of activity. The femoral portion of the sciatic nerve showed no proximo-distal bias in the distribution of phosphofructokinase activity and there were no differences in the activities of anatomically equivalent segments from contralateral nerves. Phosphofructokinase activity was subject to both anterograde and retrograde axonal transport as indicated by accumulation on both sides of a constriction applied to the sciatic trunk. Accumulation was progressive and appeared to be linear with time for at least 24 h. Linearity was lost at constrictions applied for 48 h. In experiments in which synchronous double ligations were applied to the nerve (9 mm apart), there was no redistribution of phosphofructokinase activity in the segment of nerve isolated between the two constrictions.     

拟除虫菊酯对家蝇Na-K-ATPase抑制作用的研究

通过对家蝇神经系统Na-K-ATPase性质的研究,表明Na-K-ATPase反应的适宜pH值为7.0～8.0,适温为35～40℃,K_m为0.22 mmol/L,V_max为555.56 nmol/(mg·min)。比较测定了家蝇敏感品系、Del-R、2Cl-R抗性品系的Na-K-ATPase活性及溴氰菊酯和氯菊酯对该酶的抑制作用。实验证明,敏感与抗性品系间Na-K-ATPase活力无显著的差异,但溴氰菊酯和氯菊酯对不同家蝇品系Na-K-ATPase的抑制作用有明显区别,两种拟除虫菊酯可抑制敏感家蝇品系Na-K-ATPase的活性,而对抗性品系无明显的抑制作用。     

QUININE HYDROCHLORIDE ACCEPTABILITY AFTER WATER DEPRIVATION IN GALLUS DOMESTICUS

The latency to peck and show the oral aversion response wasmeasured when 6 Brown Leghorn hens were presented with eitherwater or 0.005 M quinine hydrochloride (QHCl) after either 2,4 or 6 h water deprivation. The absence of any differences inthe latencies to peck at water with the 3 deprivation schedulesindicated no significant increase in the level of motivation,whereas, it was found that the latencies to peck at the quininesolution were significantly reduced with increasing deprivation.After 2 h deprivation the birds showed significantly longerlatencies than with water but after 6 h deprivation there wasno significant difference. By comparing the ratio values ofthe number of pecks followed by the oral aversion response dividedby the total number of pecks given to the QHCl solution it wasfound that there was a significant decrease in this ratio from2 to 6 h deprivation. It was suggested that the increase inacceptance of the quinine hydrochloride following deprivationmay be due to changes in ‘taste’ sensitivity.     

Some Physiological Characteristics of Two Tomato Cultivars, One Tolerant and One Susceptible to Tobacco Mosaic Virus

A comparison was made of physiological activity in the tomatocv. Potentate, susceptible to tobacco mosaic virus (TMV) withthat in cv. Virocross which is tolerant. Growth analysis overa 5-week period showed Potentate to have a higher relative growthrate and a higher net assimilation rate than Virocross. In reciprocalgrafts scion growth, including dry weight, was greater on Potentaterootstocks than on Virocross. Growth responses were studiedin these grafts following foliar sprays of N-6 benzyladenine(BA) and gibberellic acid (GA). The response of the scion toGA was similar in both cultivars and there was no interactionbetween GA and rootstock for scion growth. An interaction betweenstock and BA was found for leaf growth. Bleeding sap from Virocross contained more gibberellins, morecytokinins, and a lower concentration of amino compounds, especiallynon-protein acids, than Potentate. There were more gibberellinsin the roots of Potentate but no difference between shoots ofthe two cultivars. In reciprocal grafts TMV multiplied more rapidly in Virocrossscions on Potentate rootstock than on their own, but Virocrossstocks did not affect virus multiplication in Potentate scions.Spraying Potentate plants with BA reduced the rate of virusmultiplication. It is suggested that virus multiplication in the shoot may becontrolled by two distinct root factors, viz, the supply ofnative cytokinins and the amount and nature of the nitrogenouscompounds which directly influence the rate of shoot growth.     

Effect of CCK pretreatment on the CCK sensitivity of rat polymodal gastric vagal afferent in vitro

To prevent the blood-borne interference and reflex actions via neighboring organs and the central nervous system, the study was conducted in an in vitro isolated stomach-gastric vagus nerve preparation obtained from overnight-fasted, urethan-anesthetized rats. Afferent unit action potentials were recorded from the gastric branch of the vagus nerve. The left gastric artery was catheterized for intra-arterial injection. In vitro we found that 1) 55/70 gastric vagal afferents (GVAs) were polymodal, responding to CCK-8 and mechanical stimuli, 13 were mechanoreceptive, and 2 were CCK-responsive; 2) sequential or randomized intra-arterial injections of CCK-8 (0.1-200 pmol) dose-dependently increased firing rate and reached the peak rate at 100 pmol; 3) the action was suppressed by CCK-A (Devazepide) but not by CCK-B (L-365,260) receptor antagonist; 4) neither antagonist blocked the mechanosensitivity of GVA fibers. These results are consistent with corresponding in vivo well-documented findings. Histological data indicate that the layered structure of the stomach wall was preserved in vitro for 6-8 h. Based on these results, it seems reasonable to use the in vitro preparation for conducting a study that is usually difficult to be performed in vivo. For instance, because there was no blood supply in vitro, the composition of the interstitial fluid, i.e., the ambient nerve terminals, can be better controlled and influenced by intra-arterial injection of a defined solution. Here we report that acutely changing the ambient CCK level by a conditioning stimulus (a preceding intra-arterial injection of increasing doses of CCK-8) reduced the CCK sensitivity of GVA terminals to a subsequent test stimulus (a constant dose of CCK-8 intra-arterial injection).     

Measurement of sympathetic nerve activity in the unanesthetized rat

A new system was developed in our laboratory to continuously monitor intra-arterial pressure, heart rate, and sympathetic nerve activity in unanesthetized rats. The animals were prepared 24 h before the start of the experiments. Sympathoneural traffic was measured at the level of splanchnic nerve. The amplitude of the spikes recorded at this level was utilized to express sympathetic nerve activity. The amplitude of the residual electroneurogram signal present 30 min after the rats were killed was 32 +/- 2 mV (mean +/- SE; n = 11). For analysis, these background values were subtracted from values determined in vivo. The nerve we studied contains postganglionic fibers, since electrical activity decreased in response to ganglionic blockade with pentolinium (1.25 mg/min iv for 4 min). The amplitude of spikes fell by 43 +/- 4% (n = 4). Sympathetic nerve activity was highly reproducible at a 24-h interval (104 +/- 26 vs. 111 +/- 27 mV for the amplitude of spikes; n = 11). Dose-response curves to the alpha 1-stimulant methoxamine and to bradykinin were established in four rats. The increase in blood pressure induced by methoxamine caused a dose-dependent fall in sympathetic nerve activity, whereas the blood pressure reduction resulting from bradykinin was associated with a dose-dependent activation of sympathetic drive. These data therefore indicate that it is possible with out system to accurately measure sympathetic nerve activity in the awake rat, together with intra-arterial pressure and heart rate.     

Investigations of thermotropic phase changes in peripheral nerve of frog and rat. A spin label study

The temperature-dependent fluidity of myelin of frog and rat peripheral nerve (Nervus ischiadicus) was studied using the spin label technique. In frog nerve a phase change was detected at 38 °C. In rat nerve no sharp phase change could be established, and the lipid-depleted frog and rat nerve also showed no transition. From the spectral data, it was concluded that in frog and rat nerve the lipid-protein interactions are different, i.e. species dependent. Ca²⁺-depletion of frog nerve caused a loss of transition, while rat nerve remained unaffected. Thus it was indicated that, in frog nerve, Ca²⁺ is involved in the phase change. In the total lipid extract of frog nerve a phase change centered at 32 °C occurred, while the total lipid extract of rat nerve again showed no transition. It is suggested that a connexion exists between our results and investigations on the temperature dependence of an axonal conduction block of nerve.     

Histological characterization of root-knot nematode resistance in cowpea and its relation to reactive oxygen species modulation

Root-knot nematodes (Meloidogyne spp.) are sedentary endoparasiteswith a broad host range which includes economically importantcrop species. Cowpea (Vigna unguiculata L. Walp) is an importantfood and fodder legume grown in many regions where root-knotnematodes are a major problem in production fields. Severalsources of resistance to root-knot nematode have been identifiedin cowpea, including the widely used Rk gene. As part of a studyto elucidate the mechanism of Rk-mediated resistance, the histologicalresponse to avirulent M. incognita feeding of a resistant cowpeacultivar CB46 was compared with a susceptible near-isogenicline (in CB46 background). Most root-knot nematode resistancemechanisms in host plants that have been examined induced ahypersensitive response (HR). However, there was no typicalHR in resistant cowpea roots and nematodes were able to developnormal feeding sites similar to those in susceptible roots upto 9–14 d post inoculation (dpi). From 14–21 dpigiant cell deterioration was observed and the female nematodesshowed arrested development and deterioration. Nematodes failedto reach maturity and did not initiate egg laying in resistantroots. These results confirmed that the induction of resistanceis relatively late in this system. Typically in pathogen resistanceHR is closely associated with an oxidative burst (OB) in infectedtissue. The level of reactive oxygen species release in bothcompatible and incompatible reactions during early and latestages of infection was also quantified. Following a basal OBduring early infection in both susceptible and resistant roots,which was also observed in mechanically wounded root tissues,no significant OB was detected up to 14 dpi, a profile consistentwith the histological observations of a delayed resistance response.These results will be useful to design gene expression experimentsto dissect Rk-mediated resistance at the molecular level. Key words: Cowpea, histology, hypersensitive response, Meloidogyne incognita, reactive oxygen species, root-knot nematode, Vigna unguiculata Received 5 November 2007; Revised 22 January 2008 Accepted 23 January 2008     

Ultracytochemical localisation and comparison of adenyl cyclase activities in pineal bodies of Wistar rats and mongolian gerbils

Summary In the pineal bodies of Wistar rats and mongolian gerbils (Meriones unguiculati) the main concentration of adenyl cyclase was found in capillary endothelial cells, their basal laminae, pericapillary amorphous material and in nerve endings. In the intercellular clefts between the rat pineal cells there was no adenyl cyclase activity, but a considerable amount of this enzyme was detected in the gerbils. The adenyl cyclase lacks completely in the pinealocyte cytoplasms of both species. It was concluded that the gerbil pineal cells may be more reactive to various hormonal influence than those of the rat.With the technical assistance of Miss Ch. Thommen and Mr. P.-A. MilliquetDedicated to Professor Dr. med. O. Bucher on the occasion of his 65th birthday     

The cysteine-rich region of T1R3 determines responses to intensely sweet proteins

A wide variety of chemically diverse compounds taste sweet, including natural sugars such as glucose, fructose, sucrose, and sugar alcohols, small molecule artificial sweeteners such as saccharin and acesulfame K, and proteins such as monellin and thaumatin. Brazzein, like monellin and thaumatin, is a naturally occurring plant protein that humans, apes, and Old World monkeys perceive as tasting sweet but that is not perceived as sweet by other species including New World monkeys, mouse, and rat. It has been shown that heterologous expression of T1R2 plus T1R3 together yields a receptor responsive to many of the above-mentioned sweet tasting ligands. We have determined that the molecular basis for species-specific sensitivity to brazzein sweetness depends on a site within the cysteine-rich region of human T1R3. Other mutations in this region of T1R3 affected receptor activity toward monellin, and in some cases, overall efficacy to multiple sweet compounds, implicating this region as a previously unrecognized important determinant of sweet receptor function.     

A pharmacological investigation of the biphasic nature of the contractile response of rabbit and rat vas deferens to field stimulation

It has been demonstrated previously with the vas deferens of the guinea-pig that the first and second phases of the contractile response to motor nerve stimulation are preferentially antagonized by the P₂-purinoceptor antagonist arylazido aminopropionyl ATP (ANAPP₃), and the α₁-adrenoceptor antagonist prazosin, respectively. We have now investigated the effect of the two antagonists on the biphasic contraction in the vas deferens of two other species; rabbit and rat. ANAPP₃, in a concentration which antagonized responses to exogenously applied ATP but not those to exogenous norepinephrine, preferentially reduced the initial phasic response of the rabbit vas deferens to motor nerve stimulation without significantly reducing the secondary, tonic phase of the response. Prazosin had the opposite effect; antagonizing the response to norepinephrine but not to ATP and reducing the tonic response to motor nerve stimulation without significantly reducing the initial phasic response. Results obtained with the rat vas deferens were similar. The present results combined with previous findings suggest that ATP and norepinephrine act as cotransmitters in the vas deferens of several species.     

烟夜蛾和棉铃虫对高浓度烟草挥发物的电生理和行为反应

寡食性的烟夜蛾Helicoverpa assulta (Guenée)和广食性的棉铃虫H. armigera (Hübner)是夜蛾属Heliothis的两近缘种昆虫。主要的农作物中,棉花和番茄上极少发现烟夜蛾,而辣椒上几乎没有棉铃虫,只有烟草均为二者所嗜食,也唯独在烟草上它们可以稳定共存。为明确植物挥发物对昆虫寄主定向和选择行为的影响,本文运用触角电位（EAG）和风洞技术,测定了两种夜蛾的处女雌蛾、交配雌蛾及雄蛾对20种高浓度（0.1 mol/L）烟草挥发物的电生理和行为反应。结果表明：烟夜蛾和棉铃虫对测试的烟草挥发物均可产生EAG反应,而且都表现出绿叶气味＞脂类和芳香化合物＞单萜、倍半萜和杂环化合物的总体反应趋势;两种夜蛾对多数化合物的EAG反应既无显著的种内性别差异,也无显著的种间差异,表明二者嗅觉神经系统对寄主气味图谱的识别和感受能力具有一定的相似性,可对寄主植物的化学信息产生相同的“理解”;二者的行为反应结果与EAG反应结果基本一致,即两种夜蛾受到的嗅觉刺激与产生的行为反应具有统一性,结合两种夜蛾寄主范围的差异,推测烟草挥发物主要影响两种夜蛾的寄主定向行为,而寄主选择行为可能更依赖于二者与植物接触后对植物理化性质的评价过程。     

甜味蛋白thaumatin基因转入烟草的研究

利用基因工程技术 ,将分别克隆在两个不同载体上的甜味蛋白 thaum atin c DNA基因片段连接成一个完整的 c DNA基因 ,并将该基因克隆进 p BI12 1,构建成表达载体 p BI12 1- tha.通过冻融法导入农杆菌 ,农杆菌介导叶盘法转入烟草 ,经过组培 ,得到转基因的植株 .提取转基因烟草总 DNA,经 PCR,PCR- Southern和 Southern杂交证实 ,甜味蛋白基因已整合到烟草基因组中 .RT- PCR结果证明 ,thaumatin基因已在转基因烟草中转录成 m RNA,但SDS- PAGE和甜味尝试都表明 thaumatin基因在转基因烟草中没有表达出甜味蛋白