Presynaptic inhibition and the participation of GABAB receptors at neuromuscular junctions of the crab Eriphia spinifrons

Presynaptic inhibition exerted by the common inhibitor on the closer and opener muscles and by the specific inhibitor on the opener muscle was investigated in the crab Eriphia spinifrons. In the closer muscle, activation of GABA_B receptors by baclofen reduced the mean quantal content of excitatory junctional currents by about 25%. Blocking GABA_B receptors with CGP 55845 diminished presynaptic inhibition at a similar percentage. GABA_B receptor-mediated presynaptic inhibition is linked to G proteins. Application of pertussis toxin eliminated about 25% of the inhibition exerted by the common inhibitory neuron. GABA_B receptors participate in presynaptic inhibition at release boutons of the slow and the fast closer excitor at a similar percentage. In the opener muscle, presynaptic inhibition of transmitter release from the same endings of the opener excitor was about 15% stronger with the specific inhibitor than with the common inhibitor. About 10% of the presynaptic inhibition produced by either one of the two inhibitors could be abolished by blocking GABA_B receptors. The amplitudes of the excitatory junctional currents in the opener were reduced in the presence of baclofen by about 25%, suggesting that synaptic terminals of the opener excitor are endowed with a similar percentage of GABA_B receptors as terminals of the slow and the fast closer excitors. Baclofen had no effect on postsynaptic inhibition, indicating that GABA_B receptors are not involved in postsynaptic neuromuscular inhibition. Accepted: 8 January 2000     

Effect of sex pheromone emission on the attraction of Lobesia botrana

Since the discovery of Lobesia botrana Denis & Schiffermüller (Lepidoptera: Tortricidae) sex pheromone, it has played an important role in the control and detection of this pest, for example, through the use of pheromone‐baited traps and mating disruption techniques. Rubber septa are the most common pheromone dispensers used in monitoring traps, but often dispenser performance is not optimized. The key to improve methods based on pheromones as attractants (monitoring, mass trapping, or ‘attract and kill’) is to know the optimum emission interval, because release rates can strongly affect the attraction. In this work, five levels of pheromone load with different release rates were compared in traps using mesoporous pheromone dispensers to investigate the optimum release rate maximizing L. botrana catches. Residual pheromone loads of the dispensers were extracted and quantified by gas chromatography, to study release profiles and to estimate the various emission levels. The efficacy of pheromone emission was measured in field trials as number of moths caught. A quadratic model was fitted to relate the numbers caught vs. the daily emission rates. The resulting quadratic term was statistically significant, confirming the existence of a relative maximum for L. botrana catches. Taking into account that the trial was carried out only in one location, an optimum emission value of ca. 400 μg per day could be considered to enhance the attraction of L. botrana under West‐Mediterranean weather conditions.     

Experimental release of an Iberian lynx (Lynx pardinus)

Reintroduction to the wild of threatened species has become a modern additional justification for captive propagation. This conservation procedure is costly, and both economic resources and the absence of optimal conditions in the field limit the IUCN recommendations for reintroduction to a small proportion of potential candidate species. Furthermore reintroduction attempts often fail. In carnivores, reintroduction failure is attributed to unsuitable adaptation in the field by captive-reared animals, due to their lack of hunting skills, their tendency to leave the target area, their inadequate interaction with conspecifics or their excessive confidence in humans. This list of causes is based on very few studies of carnivore adaptation after reintroduction. In very rare and endangered species, monitoring individual case-histories is the only way to evaluate reintroduction success. We report a successful experimental release of an Iberian lynx (Lynx pardinus) which grew up in captivity. Careful feeding-training and avoidance of human contact during the captive phase, as well as good habitat quality and correct interaction with other wild lynx in the release site, seem to account for the observed success. Permanence of the lynx within the release area might be related to the availability of territory vacancies in the receiving population. Our results allow some optimism for future reintroductions of this endangered species in areas where it has become extinct recently.     

Gastrin and the ultrastructure of G cells after stimulation with acetylcholine

The effect of intragastric administration of acetylcholine on serum and antral gastrin concentrations of rats has been examined using a radioimmunoassay and quantitative electron microscopy. Exposure of the stomach of rats, previously fasted for 24h, to 2% acetylcholine for either 0.5 or 2h resulted in a significant 4--5 fold increase in serum gastrin concentrations to levels similar to those found in fed animals. Such treatment produced no detectable change in antral gastrin concentration or in the number or electron density of secretory granules in the G cells. This lack of detectable change in the G cells was not unexpected since our calculations suggest that less than 10% of the total gastrin stored in the antrum is released over 2h as a result of the stimulation with acetylcholine. The proportion of electron-lucent secretory granules was, however, markedly increased by prolonged fixation in aldehydes. The increase was similar in both ACh stimulated and control animals. These results indicate that the ultrastructural appearance of G cell secretory granules in influenced far more by the conditions of fixation than by the release of gastrin. They therefore cast considerable doubt on the hypothesis that gastrin is released by molecular dispersion from the granules.     

Hydrogen peroxide release from human blood platelets

The release of hydrogen peroxide from human blood platelets after stimulation with particulate membrane-perturbing agents has been determined by fluorescence using scopoletin as the detecting agent. Platelet suspensions containing less than 1 polymorphonuclear leukocyte/10⁸ platelets showed a significant release of hydrogen peroxide (6.11 nmol/10⁹ platelets per 20 min, S.D., 0.26, n=9) after addition of zymosan or latex particles, compared to unstimulated platelets. The release of hydrogen peroxide was only observed when the scopoletin was added to the platelet suspensions during the stimulation. Any attempt to determine hydrogen peroxide release in the supernatant at the end of the incubation with zymosan or latex failed. A NADH-dependent production of hydrogen peroxide was observed by measuring the difference of oxygen uptake in the presence and absence of catalase (500 units), which was not inhibited by potassium cyanide (1 mM). By this method the NADH-dependent cyanide-insensitive peroxide production and release was 6.0 nmol/10⁹ platelets per 20 min from resting platelets (S.D., 2, n=6) vs. 15 nmol/10⁹ platelets per 20 min from stimulated platelets (S.D., 2, n=6).     

Effects of methylglyoxal on platelet hydrogen peroxide accumulation,aggregation and release reaction

Methylglyoxal generates a slight increase in the basal level of hydrogen peroxide in platelets. The oxidation effect of methylglyoxal significantly potentiated by thrombin, depends on both the ketoaldehyde and the agonist concentrations. A further significant increase in hydrogen peroxide accumulation was obtained in platelets pretreated with the alkylating agent N-ethylmaleimide which depletes GSH and blocks glutathione peroxidase. Resting platelets completely transform the ketoaldehyde into D (?)lactate, whereas stimulated platelets transform about 10–15 per cent of the metabolized methylglyoxal into D (?)lactate. The metabolic modifications generated by methylglyoxal such as the GSH depletion and hydrogen peroxide accumulation induce modifications in platelet function. Methylglyoxal inhibits platelet aggregation induced by several agonists and ATP release induced by thrombin.     

Pseudopodia formation by neurosecretory granules

Summary Ultrastructural studies of the mouse neurohypophysis, under various experimental conditions, revealed a number of neurosecretory granules (NSG) bearing single pseudopodia-like protrusions. Some NSG adhered to the axolemma via pseudopodia; other NSG, distant from the axolemma, budded electron lucent microvesicles from the tip of the pseudopod.Pseudopodia counts were made on electron micrographs, and calculated as a percentage of the NSG population. In neural lobes from intact mice, small numbers of pseudopodia were observed (0.3%); the count increased significantly after injections of large doses of horseradish peroxidase (HRP) (9.4–14.5%); hypertonic saline augmented the count, as did histamine.In vitro incubation experiments with isolated neural lobes in Krebs Ringer revealed concomitant pseudopodia formation and elevated vasopressin release (measured by antidiuretic bioassay) in the presence of HRP and di-butyryl cyclic AMP respectively. Histamine and excess potassium also increased hormone secretion, but did not induce pseudopodia formation in vitro; pseudopodia were observed neither in controls, nor in the presence of ineffective secretagogues.It is suggested that the pseudopod may represent the active site on the granule membrane. Different ultrastructural images of granule release suggest that several modes of hormone release may be operative in the neurohypophysis. The role of HRP in pseudopodia formation and vasopressin release is enigmatic.This study is dedicated to Prof. Berta Scharrer, esteemed mentor and beloved friendProf. Zwi Selinger, Department of Biological Chemistry of the Hebrew University, kindly collaborated in the in vitro experiments. Thanks are due to Mrs. Ilana Sabnai and Mrs. Sara Eimerl for excellent technical assistance. Research supported by the Binational Israel-United States Science Foundation (BNSF), grant 200     

Effects of gonadotropin-releasing hormone agonist administered in microparticles on sperm quality and quantity,and plasma sex steroid levels in northern pike

Artificial reproduction of northern pike Esox lucius is impeded by the likelihood of obtaining only a small volume of sperm of inconsistent quality. A controlled-release hormone delivery system has the potential to enhance sperm production while avoiding multiple injections The objective of this study was to investigate the effects of mammalian gonadotropin-releasing hormone agonist (mGnRHa) incorporated into poly(lactic-co-glycolic acid) (PLGA) microparticles on milt production, spermatozoon characteristics, and secretion of 17β-estradiol (E2), 11-keto testosterone (11-KT), and testosterone in northern pike. Fish were divided into four groups and injected with 2 mg/kg BW carp pituitary extract (CPE), 20 µg/kg BW mGnRHa in PLGA microparticles, or 20 µg/kg BW mGnRHa plus 20 mg/kg BW metoclopramide (MET) in PLGA microparticles (PLGA + MET), along with a control group injected with 1 ml/kg 0.9% NaCl. At 48 h postinjection, the volume of milt produced was significantly greater in groups treated with CPE and PLGA + MET than in other groups. At 96 h postinjection, all hormone-treated groups exhibited significantly higher spermatozoon average velocity than recorded in the control group. Spermatozoon motility was significantly increased (P < 0.05) in the CPE and PLGA groups compared to baseline values. All treated groups showed significantly lower levels of 11-KT after the hormone injection compared to baseline values and to controls. Plasma testosterone levels increased in all hormone-treated groups. The use of PLGA microparticles, with or without metoclopramide, is suitable for use as a carrier of hormone treatments to regulate spermiation in mature northern pike.     

Warming and grazing enhance litter decomposition and nutrient release independent of litter quality in an alpine meadow

增温和放牧对高寒草甸凋落物分解及其养分释放的影响不依赖于凋落物品质在放牧生态系统中,增温、放牧和凋落物品质共同决定着凋落物分解和养分释放。然而,在以往的研究中这些因子的效应通常被单独地研究。在本研究中,我们在青藏高原高寒草甸开展了一个昼夜非对称增温和中度放牧两因子的凋落物分解试验。从每个处理中收集了凋落物样品,这些凋落物一部分放在它们的来源处理小区,另一部分放在其他处理小区以此来探究增温、放牧以及凋落物品质对凋落物分解和养分释放的影响。研究结果表明,增温而不是放牧显著增加了凋落物质量的损失、单位面积全碳、全氮以及全磷含量的损失,这主要是因为增温增加了凋落物生物量和分解速率。然而,尽管同时增温放牧处理也加快了凋落物分解速率,但由于降低了凋落物生物量,所以增温放牧处理并没有显著影响单位面积的凋落物碳和养分释放量。相比木质素含量和碳氮比而言,季节性土壤平均温度能够更好地预测凋落物分解速率。增温和放牧对凋落物分解存在交互作用,但它们和凋落物品质对凋落物的影响均不存在交互作用。单位面积的总氮释放的温度敏感性要高于总磷。因此,我们的结果表明,增温对凋落物分解以及养分释放的影响要显著大于凋落物品质变化对其分解的影响。在高寒草甸,氮释放的增加可能会间接导致土壤磷有效性的缺乏。     

不同施肥条件下毛叶苕子的腐解及养分释放特征

利用田间埋袋法,研究不施肥、施氮肥、施石灰3种处理对豫南稻田毛叶苕子腐解及养分释放特征的影响.结果表明:不同施肥处理下毛叶苕子累积腐解率为65.3%~72.5%,腐解过程中呈现前11 d腐解较快、后期腐解缓慢并逐渐趋于平稳的趋势.不同处理养分释放率表现为钾>磷>碳>氮,试验结束时(翻压148 d),碳、氮、磷、钾的累积释放率分别为83.6%~84.6%、78.2%~81.2%、89.8%~91.4%、96.3%~97.0%.在整个腐解期内,毛叶苕子氮释放特征与腐解特征相似,与不施肥相比,施石灰促进毛叶苕子腐解及氮、磷、钾养分释放;施氮肥促进毛叶苕子磷释放,抑制钾释放;施石灰和氮肥对碳释放均无显著影响.施氮肥处理腐解0~11 d促进毛叶苕子腐解及氮释放,腐解11~148 d抑制毛叶苕子腐解及氮释放.采用一级动力学方程及对数函数方程拟合豫南稻区毛叶苕子腐解及碳、氮、磷、钾养分释放特征均达到显著水平,拟合方程的特征参数值与毛叶苕子腐解率及养分释放率呈显著相关.施用石灰促进毛叶苕子腐解及养分释放的效果优于施用氮肥;一级动力学方程及对数函数方程特征参数值可较好地描述毛叶苕子腐解及养分释放能力.