Analysis of connexin expression during seizures induced by 4-aminopyridine in the rat hippocampus

Background

In epilepsy, seizures are generated by abnormal synchronous activity in neurons. In the rat hippocampus (HIP), epileptiform activity has been found to be associated with gap junctions (GJs). GJs are formed by the combination of two hemichannels, each composed of six connexins. At low doses, the convulsive drug 4-aminopyridine (4-AP) produces epileptiform activity without affecting glutamate levels; therefore, GJs could participate in its effect. Based on this argument, in this study, the expression of Cx 32, Cx 36 and Cx 43 protein and mRNA in the HIP of rats treated with 4-AP was evaluated. The evaluation of connexins was carried out by chemifluorescent immunoassay, semiquantitative RT-PCR and immunofluorescence to detect the amount and distribution of connexins and of cellular markers in the HIP and dentate gyrus (DG) of animals treated with NaCl and 4-AP in the right entorhinal cortex. In these animals, convulsive behavior and EEG signals were analyzed.

Results

The animals treated with 4-AP showed convulsive behavior and epileptiform activity 60 min after the administration. A significant increase in the protein expression of Cx 32, Cx 36 and Cx 43 was found in the HIP contralateral and ipsilateral to the site of 4-AP administration. A trend toward an increase in the mRNA of Cx 32 and Cx 43 was also found. An increase in the cellular density of Cx 32 and Cx 43 was found in the right HIP and DG, and an increase in the cellular density of oligodendrocytes in the DG and a decrease in the number of cells marked with NeuN were observed in the left HIP.

Conclusions

Cx 32 and Cx 43 associated with oligodendrocytes and astrocytes had an important role in the first stages of seizures induced by 4-AP, whereas Cx36 localized to neurons could be associated with later stages. Additionally, these results contribute to our understanding of the role of connexins in acute seizures and allow us to direct our efforts to other new anticonvulsant strategies for seizure treatment.     

Increased hippocampal noradrenaline is a biomarker for efficacy of vagus nerve stimulation in a limbic seizure model

Vagus nerve stimulation (VNS) is an effective adjunctive treatment for medically refractory epilepsy. In this study, we measured VNS-induced changes in hippocampal neurotransmitter levels and determined their potential involvement in the anticonvulsive action of VNS, to elucidate the mechanism of action responsible for the seizure suppressing effect of VNS in an animal model for limbic seizures. We used in vivo intracerebral microdialysis to measure VNS-induced changes in hippocampal extracellular concentrations of noradrenaline, dopamine, serotonin and GABA in freely moving, male Wistar rats. During the same experiment, the effect of VNS on pilocarpine-induced limbic seizures was assessed using video-EEG monitoring. The involvement of VNS-induced increases in hippocampal noradrenaline in the mechanims of action of VNS was evaluated by blocking hippocampal α(2)-receptors. VNS produced a significant increase in hippocampal noradrenaline concentration (69 ± 16% above baseline levels). VNS also increased the latency between pilocarpine infusion and the onset of epileptiform discharges, and reduced the duration and severity of pilocarpine-induced limbic seizures. A strong positive correlation was found between the noradrenergic and anticonvulsive effects of VNS. Blockade of hippocampal α(2 -receptors reversed the seizure-suppressing effect of VNS. VNS induces increases in extracellular hippocampal noradrenaline, which are at least partly responsible for its seizure-suppressing effect in a model for limbic seizures, and constitute a potential biomarker for the efficacy of VNS in temporal lobe epilepsy.     

Chasing fate and function of new neurons in adult brains

Neuron production, migration and differentiation are major developmental events that continue, on a smaller scale, into adult life in a wide range of species from insects to mammals. Recent reports of adult neurogenesis in primates, including humans, have led to explosive scientific and public attention. During the last two years, significant discoveries have revealed that the generation, recruitment and survival of new neurons in adult brains are governed by principles similar to those that shape the developing brain, such as neuronal death, sensory experience, activity levels, and learning. Similarly, many factors implicated in embryonic neurogenesis are increasingly found to regulate adult neurogenesis and survival as well. These findings now allow the first manipulations of the numbers of adult-generated neurons to address their potential behavioral function.     

Allocating time between feeding, resting and moving by the two-spotted spider mite, Tetranychus urticae and its predator Phytoseiulus persimilis

This study characterizes the timing of feeding, moving and resting for the two-spotted spider mite, Tetranychus urticae Koch and a phytoseiid predator, Phytoseiulus persimilis Athias-Henriot. Feeding is the interaction between T. urticae and plants, and between P. persimilis and T. urticae. Movement plays a key role in locating new food resources. Both activities are closely related to survival and reproduction. We measured the time allocated to these behaviours at four ages of the spider mite (juveniles, adult females immediately after moult and adult females 1 and 3 days after moult) and two ages of the predatory mite (juveniles and adult females). We also examined the effect of previous spider mite-inflicted leaf damage on the spider mite behaviour. Juveniles of both the spider mite and the predatory mite moved around less than their adult counterparts. Newly emerged adult female spider mites spent most of their time moving, stopping only to feed. This represents the teneral phase, during which adult female spider mites are most likely to disperse. With the exception of this age group, spider mites moved more and fed less on previously damaged than on clean leaves. Because of this, the spider mite behaviour was initially more variable on damaged leaves. Phytoseiulus persimilis rested at all stages for a much larger percentage of the time and spent less time feeding than did T. urticae; the predators invariably rested in close proximity to the prey. Compared to adult predators, juveniles spent approximately four times as long handling a prey egg. The predator-prey interaction is dependent upon the local movement of both the predators and prey. These details of individual behaviours in a multispecies environment can provide an understanding of population dynamics.     

The adaptive significance of behavioural ontogeny in some. centrarchid fishes

Synopsis Studies on the ontogeny of behaviour in fish have seldom considered the adaptive significance of the order of appearance of the behaviours. Results of laboratory studies and field observations on the ontogeny of feeding, predator-avoidance, and agonistic behaviour in largemouth bass, Micropterus salmoides, rock bass, Ambloplites rupestris, and pumpkinseed, Lepomis gibbosus, young indicate that the order of appearance of the various behavioural systems enhances the survival of the young. In the laboratory, all species spend significantly more time feeding during the first weeks of free-swimming than the later weeks. During a corresponding period in the field the young are either occupying an offshore area low in predators (rock bass, pumpkinseed) or are being guarded by a parent (largemouth bass); thus the risk of predation is low. When the young are in a predator-rich environment (inshore) both the predator-avoidance response and agonistic behaviour are well developed. Agonistic behaviour is the last to appear and may serve to disperse the young. Dispersal may relate to the feeding mode of the various species and may also reduce the probability of predation. It is apparent that the sequential onsets of the behavioural systems are in concert with ecological events and selective pressures confronting centarchid young.     

Consequences of competition on the time budgets,growth and distributions of three species of terrestrial slugs

Summary The behavioural time budgets of two non-aggressive slugs and one aggreessive species were evaluated in monocultures or in 2-way combinations in field cages. The seasonal time budgets ofL. maximus, A. ater andA. columbianus in monoculture were remarkably similar despite their strong differences in physiological resource allocation. Similarity in physical design and resource utilization appeared to be more important determinants of behaviour. The time budgets showed distinct seasonal patterns associated with underlying physiological demands for reserve accumulation, sperm acquisition and oviposition.A. ater andL. maximus displayed a definite dispersal phase associated with femalephase maturation. In combination,A. ater andA. columbianus had no detectable effect on one another’s time budgets.L. maximus disrupted nearly every category of behaviour in these non-aggressive species, but its own time budget was not significantly different in the monoculture or in the two combinations, except for minor effects on resting and sexual behaviour.L. maximus appeared to be adapted to maximize behavioural disturbance of interspecific competitors while minimizing intraspecific effects.     

A behavioural scale to measure ease of farrowing in sows

Farrowing is one of the most critical phases in pig production, as it has an impact on neonatal pig survival. Assessing the ease of farrowing can improve the management of sows and thus increase litter survival. The aims of this study were: 1) to develop an ease of farrowing score (EFS) in sows based on the behaviour of the sows and their piglets, and 2) to determine the relationship between the EFS and productive, physiological, and subjective parameters. Eighty hybrid (Large White × Landrace) sows from first to seventh parity housed in individual crates were used. An EFS was constructed using the total duration of farrowing, the birth interval, the total time standing or sitting, the number of position changes during the day before and the day of farrowing, the sow posture at birth, the viability and the position of the piglets at birth (head or back born). Moreover, rectal temperature at 90 min after farrowing, a four categorical subjective visual assessment (VA) of farrowing and litter size (piglets born alive, stillborn and mummified foetus) were recorded. A common factor analysis model yielded five factors with an eigenvalue higher than 0.95 that accounted for 75.05% of the total variation between individuals. The three main factors were “farrowing duration”, “sow posture”, and “sow activity” of sows and explained 23.44%, 15.67%, and 14.23% of the variance, respectively. Primiparous sows had higher values for factor 3 (sow activity) than multiparous sows (P = 0.02). Sows without stillborn or mummification foetus showed higher values of EFS than sows with at least one stillborn or mummification foetus (P = 0.06 and 0.01, respectively). Sows that received a visual assessment of 3 and 4 showed higher values of EFS than sows that received a VA of 1 and 2 (P = 0.0017). The EFS appears to be a good behavioural scale to measure ease of farrowing in sows kept in individual farrowing crates. Duration of farrowing, sow position, and presence of stillborn piglets and mummified foetuses appear to be important ease of farrowing indicators.     

两种海马鳃组织和消化道的黏液细胞类型及分布

运用阿新兰(AB,pH 2.6)和过碘酸雪夫氏(PAS)反应染色方法,对三斑海马(Hippocampus trimaculatus)和日本海马(H.japonicus)鳃组织与消化道中的黏液细胞类型及分布进行了研究。染色结果显示:两种海马的鳃组织和消化道中均含有黏液细胞,日本海马的鳃组织中含有Ⅰ型和Ⅳ型黏液细胞,三斑海马的鳃组织中含有Ⅰ型、Ⅲ型和Ⅳ型黏液细胞。两种海马消化道各部位的黏液细胞类型和数量有明显差异:日本海马的食道中Ⅰ型细胞最多,而三斑海马的食道中Ⅳ型细胞最多;日本海马的前肠中只含有Ⅰ型细胞,而三斑海马的前肠中含有Ⅰ型、Ⅲ型和Ⅳ型细胞,其中Ⅰ型细胞含量最多;日本海马的中肠中含有Ⅰ型、Ⅲ型和Ⅳ型细胞,其中Ⅲ型细胞含量最多,而三斑海马中肠中只含有Ⅰ型细胞;日本海马与三斑海马的后肠中都分布有Ⅰ型、Ⅱ型、Ⅲ型和Ⅳ型细胞,两者不同的是,日本海马的后肠中Ⅲ型细胞含量最多,三斑海马的后肠中Ⅳ型细胞含量最多。     

一种家兔持续静脉麻醉方法

目的为进行某些特殊项目的实验研究,建立一种持续麻醉实验动物模型。方法戊巴比妥钠粉剂加生理盐水溶解,配成1%水剂,按3 mL/kg（30 mg/kg）耳缘静脉注射。诱导成功后约2 h经留置针使用恒速微量泵1 mL/h.kg给药,维持4 h后调节至2 mL/h.kg给药,并长期维持。每日上午、下午各停止静脉给药1 h,为动物饲食水。结果经实验证明家兔可耐受120 h连续麻醉。实验动物耐受情况好,家兔麻醉前呼吸48～68次/min,心跳140～200次/min,体温为39.4℃±1.0℃。以3～6 mg/s速度,成年兔的正常耳缘静脉注射后,平均3＋2.8 s实验动物呈三度麻醉状态,无自主运动,未见兴奋躁动过程,呼吸24±6.6次/min。死亡率12.5%。持续麻醉期间动物二便正常。结论长期持续麻醉的效果要求是以避免动物挣扎的镇静为主,戊巴比妥适合于建立长期麻醉实验动物模型。该方法在爆炸伤和创面愈合等实验实际应用中效果理想。     

Effect of small doses of naloxone on sexual exhaustion in White New Zealand male rabbits

The objective of the present study was to determine the effect of small doses of naloxone on sexual exhaustion in White New Zealand male rabbits. Twelve young and 12 adult male rabbits 6–12 months old and 14–20 months of age, respectively, were selected from a commercial farm. Each male rabbit was housed individually in galvanized cages (90 cm × 60 cm × 40 cm). The rabbits were housed in an open shed exposed to natural photoperiod (12 L 12 D, 19°N). Daily temperature fluctuated through the year from 28 to 16 °C. Humidity was 45 ± 5%. Water and food (rabbit chow PMI) was supplied ad libitum. After sexual behaviour for each studied group was established, the males were given a 6-day rest, and 3 days before next trial, six males of each group (treated) received a subcutaneous implant of 8 mg of naloxone in a crystalline nitrocellulose pellet formulated to be completely absorbed in 15 days. The remaining six males were sham-treated (control). At the end of the resting period as previously described, the sexual behavior of each group was studied and compared using a Mann–Whitney statistical U-test. The effect of naloxone on sexual behavior was analyzed with a Wilcoxon test for correlated samples. With regard to sexual activity between young and adult rabbits, it was observed that there was a significant difference between groups (P = 0.00275, Z = 2.8823, adjusted Z = 2.99.43) showing that younger rabbits mounted/ejaculated from 9 to 10 females compared with 6 to 8 mounted/ejaculated by older rabbits. When naloxone was administered to both groups, there was a significant difference when comparing sexual behavior before and after administration of naloxone (table first and second trial). Young rabbits treated with naloxone mounted/ejaculated 11–12 females while older rabbits mounted nine females before reaching sexual exhaustion. A significant difference was observed when comparing the number of estrous females that were mounted/ejaculated between groups. Environmental photoperiod and temperature changes were not considered. It was concluded that endogenous opioids are important modulators of behavioral and hormonal interactions related to sexual behavior.     

Genetic and environmental factors affecting the suitability of dogs as Guide Dogs for the Blind

Summary Many dogs are found to be unsuitable for training as guide dogs for the blind. Consequently the Royal Guide Dogs for the Blind Association of Australia has embarked on a breeding program to produce a strain of labrador dogs which is suitable for guide dog training.The most common reasons for rejecting dogs are fearfulness, dog distraction, excitability, health and physical reasons and hip dysplasia. The selection program seems to have been successful in improving the success rate mainly by lowering fearfulness, but there has not been a continuing improvement. This is probably due to continual introduction of dogs from other populations into the breeding program.Males suffer from a higher rejection rate due to dog distraction and a lower rejection rate due to fearfulness and excitability than females, so that there is little sex difference in overall success rate.The heritability of success (0.44) is high enough to predict further progress from selection, again mainly against fearfulness.Variation in environment prior to 6 weeks of age, in age when dogs were placed into a private home and in age when males were castrated, had little effect on the success rate.     

Do disrupted early attachments affect the relationship between guide dogs and blind owners?

The study investigated the affectional bond developed by dogs (Canis familiaris) towards their human companions during the selection process to become guide dogs and compared this bond with that formed by pet dogs with their owners. One hundred and nine dog-owner pairs were tested using a modified version of the Strange Situation Test: custody dogs-puppy walkers (n = 34), apprentice dogs-trainers (n = 26), guide dogs-blind owners (n = 25) and pet dogs-owners (n = 24).

Twenty-six behaviours were scored using a 5 s point sampling method and two vocal behaviours were recorded as bouts. Factor analysis carried out on 24 mutually exclusive behaviours highlighted two different profiles of response. A relaxed reaction characterised by a high play activity was distinctive of custody and apprentice dogs, whereas an anxious reaction characterised by a high degree of proximity seeking behaviours was distinctive of pet dogs. Guide dogs were intermediate between these two extremes, expressing their attachment to the owners but showing a more controlled emotional reaction. This finding suggests that guide dogs can be viewed as “working pets”. Furthermore, the experimental set-up, characterised by the presence of a frightening stimulus, revealed that untrained dogs (pets and custody dogs) were more fearful than trained dogs (guide dogs and apprentice dogs). Finally, differences in temperament emerged between retrievers: Golden retrievers showed a higher level of affection demand while Labrador retrievers were more playful. Overall, these findings show that in spite of separations from previous attachment figures, guide dogs established with their blind owner a rather good and secure affectional bond.     

兔支气管败血波氏杆菌PCR检测方法的建立及应用

目的建立一种简便、快速、敏感、特异的适用于支气管败血波氏杆菌的PCR检测方法。方法根据兔支气管败血波氏杆菌（Bordetella bronchiseptica）的fim2基因序列设计了一对特异性引物,进行PCR扩增、特异性和敏感性试验,并将其应用于临床样品的检测。结果利用该PCR方法扩增出425bp的目的基因片段,该产物序列与GeneBank上公布的基因序列同源性为100％。特异性试验表明,该方法对大肠埃希氏菌、多杀性巴氏杆菌、魏氏梭菌和金黄色葡萄球菌均无交叉性反应;并且最小可检出菌液浓度为3．6CPU。用该PCR方法检测了从江苏、山东等地采集的146份兔鼻拭子,结果检出支气管败血波氏杆菌阳性92例,阳性率为63．01％。结论建立了快速检测支气管败血波氏杆菌的PCR方法。     

Melanoma Prevention: Is It Possible to Change a Population's Behavior in the Sun?

With the highest incidence and mortality rates of melanoma in the world, Australia has been running carefully designed primary prevention programs for more than 10 years. These have included public education programs aimed at all ages but concentrating particularly on young people. The programs have also included structural changes such as provision of shade in open space, rescheduling work and sport activities, removal of sales tax from approved sunscreens, and provision of cheap sunscreens. There have been substantial changes in knowledge, attitudes, and beliefs about melanoma, sunlight, and suntans. There have been significant increases in sun protective behavior reflected in the reduced proportions of people sunburnt at the weekend in summer in recent years. The incidence rate of melanoma continues to rise, as expected. Although there has been considerable success so far, we still have a long way to go.