Dose-response effects of orexin-A on food intake and the behavioural satiety sequence in rats

Although intracerebroventricular (i.c.v.) administration of orexin-A has been reported to stimulate food intake and/or feeding behaviour in rats, mice and goldfish, little attention has thus far been paid to its effects on normal patterns of feeding. In the present study, a continuous monitoring technique was used to characterise the effects of this novel neuropeptide on the microstructure of rat behaviour during a 1-h test with palatable wet mash. Particular attention was devoted to the behavioural satiety sequence, in which feeding is followed by grooming and resting. Although results confirmed the hyperphagic effects of orexin-A (3.33-30.0 microg i.c. v.), gross behavioural analysis failed to reveal any reliable effects of peptide treatment on eating, drinking, sniffing, grooming, resting, locomotion or rearing. However, microstructural analysis revealed behavioural effects of orexin-A that are both dose- and time-dependent. At lower doses (3.33-10.0 microg), orexin-A primarily delayed behavioural satiety, i.e. the normal transition from eating to resting. In contrast, the 30 microg dose initially induced a sedative-like effect, significantly suppressing eating and other active behaviours for the first 15-20 min of the test period. This sedative-like effect resulted in a phase-shifting of the entire behavioural sequence with higher than control levels of eating, grooming, locomotion, rearing and sniffing observed over the second half of the test session. Present findings illustrate the advantages of microstructural behavioural analysis and suggest that the hyperphagic response to low doses of orexin-A results largely from a delay in behavioural satiety while that seen in response to high doses may occur in rebound to initial behavioural suppression. Further studies will be required to confirm the identity of the specific orexin receptors (i.e. OX(1) or OX(2)) involved in mediating the dose-dependent behavioural effects reported.     

Validation of a new system for the automatic registration of behaviour in mice and rats

A newly developed behaviour registration system, Laboratory Animal Behaviour Observation, Registration and Analysis System (LABORAS) for the automatic registration of different behavioural elements of mice and rats was validated. The LABORAS sensor platform records vibrations evoked by animal movements and the LABORAS software translates these into the corresponding behaviours. Data obtained by using LABORAS were compared with data from conventional observation methods (observations of videotapes by human observers). The results indicate that LABORAS is a reliable system for the automated registration of eating, drinking, grooming, climbing, resting and locomotion of mice during a prolonged period of time. In rats, grooming, locomotion and resting also met the pre-defined validation criteria. The system can reduce observation labour and time considerably.     

Differences in response to anaesthetics and analgesics between inbred rat strains

Differences in response to analgesic and anaesthetic drugs can partly be attributed to variations in the genetic background of experimental animals. This study was carried out to determine differences in the response of inbred rat strains to a selection of analgesics and drugs used in anaesthetic protocols. A cross between the most contrasting strains can then be phenotyped in future studies in order to localize quantitative trait loci (QTLs) involved in analgesic/anaesthetic drug sensitivity. Eight inbred strains (n = 6 rats/strain) were selected for the study: the pigmented ACI, BN and COP strains and the albino F344, LEW, SHR, WAG and WKY strains. Each rat was injected intravenously with two analgesics (buprenorphine 0.05 mg/kg and nalbuphine 1 mg/kg) and three drugs used in anaesthetic protocols (propofol 25 mg/kg, medetomidine 50 microg/kg and ketamine 10 mg/kg), respectively, using a crossover design. Analgesic responses were assessed using an analgesiometric procedure. The sleep time of the rat and, where applicable, the interval between injection and loss of righting reflex were used to determine the anaesthetic response. Six out of eight strains responded significantly different from each other to the analgesic effect of buprenorphine with the ACI strain as hyper-responder. The tail withdrawal latency at 55 degrees C of the F344 and WKY rats using buprenorphine was not significantly different from baseline tail withdrawal latencies. In this study, all strains were non-responsive to the analgesic effects of nalbuphine. The response to all three drugs used in anaesthetic protocols differed significantly among the strains. The F344 and BN strains were relatively resistant to the sedative effects of medetomidine. Use of ketamine was abandoned in the ACI and BN strains when the first two animals of both strains died soon after induction. With all three drugs the sleep time of albino rats was significantly longer compared with that of the pigmented ones. We conclude that the results from this study can be used in future studies where QTLs for the sensitivity to anaesthetic/analgesic drugs are localized.     

Bombesin-induced behavioural changes: Antagonism by neuroleptics

Adult male Sprague-Dawley rats were administered bombesin (BN) intracerebroventricularly (ICV), at a dose of 0.001, 0.01, 0.1, or 1.0 μg, and the behavioural effects monitored longitudinally across time for up to 24 hr. Administration of BN significantly increased the locomotor, rearing and grooming activity at all doses. The time-course of behavioural activation was dose-related (lasting up to 2.5 hr). There was no significant difference in the behavioural response of rats receiving the BN doses in an ascending or descending order. To test the effects of dopamine receptor blockade on the BN-induced behavioural changes, groups of animals were treated with fluphenazine or haloperidol (0.1 to 2.5 mg/kg, IP) 30 min prior to BN (1 μg, ICV) administration. The results revealed that the neuroleptics could effectively antagonize the BN-induced activation of locomotor, rearing and grooming activity. These data are concordant with the view that centrally administered BN stimulates spontaneous exploratory and grooming behaviours in rats, in a time- and dose-related manner. Furthermore, since neuroleptics block these effects, it remains possible that the BN-induced behavioural changes may be mediated, at least in part, through the dopaminergic system(s).     

The behaviour of rats selected for their voluntary ethanol consumption

New strains of rats, preferent (HAP) and non-preferent (NAP) for ethanol were selectively outbred from a Wistar stock. The strains have now been raised to the F13 generation. The F9/10 animals, selected for this behavioural investigation, exhibited a significant phenotypic drinking behaviour and/or ethanol consumption. During a free choice between tap water and 10% ethanol solution (v/v), the mean daily alcohol intake for male and female HAP rats was 8.42 +/- 0.69 g/kg/24 h (n = 20 o) and 11.50 +/- 0.42 g/kg/24 h (n = 20 o), for male and female NAP rats 0.74 +/- 0.09 g/kg/24 h (n = 20 o) and 1.76 +/- 0.20 g/kg/24 h (n = 20 o), respectively. The NAP rats exhibited a strong aversion to the 10% ethanol solution when it was the only source fluid. In the open-field test (OFT), as compared to the NAP rats, male individuals of the HAP strain showed a lower motility in the first minute, in penetration into the inner squares, showed a longer latency to start exploration (latency to leave the center), exhibited larger rearing and grooming activity and shorter latencies to start these activities. The defecation rate was smaller and latency to defecation prolonged. Female HAP rats showed higher activity scores in penetration of outer and inner squares and a shorter latency to start exploration. They also had higher rearing but smaller grooming activity. The females exhibited identical defecation but different urination behaviour in comparison to the males. The time-to-emerge latencies of HAP rats were longer than in NAP individuals.     

Behavioural studies of MHC‐congenic mice

Behavioural studies of MHC‐congenic mice and rats have focused primarily on mate choice and the ability to discriminate between strains by their urine odours, but these strains may differ in other behaviours, such as activity and ultrasonic vocalizations. Ivanyi (1978, Proc. Roy. Soc. Lord. 202, 117–158) has reviewed the physiological differences associated with the MHC, many of which could influence behaviour. We have started a systematic study of behavioural development and adult behaviour in MHC‐congenic mice. A developmental test battery (growth, rate, locomotion, grooming, eye opening, ultrasonic vocalizations, etc.) was used to examine differences between C57BL/6J vs. B6‐H‐2^bm1 and C57BL/10SnJ vs. B10.BR/sgSnJ mice. A test battery of spontaneous behaviours (activity, exploration, ultrasonic vocalizations, etc.) was used to examine behavioural differences between adult C57BL/6J vs. B6‐H‐2^bm1 and C57BL/10SnJ vs. B10.BR/sgSnJ mice. Differences in development and in adult behaviours between these MHC-congenic strains is discussed in relation to possible neural, endocrine and immune system differences. Future studies will compare MHC-congenic mice on levels of anxiety, sociosexual behaviour and on learning paradigms. This revised version was published online in July 2006 with corrections to the Cover Date.     

Species specific behavioural patterns (digging and swimming) and reaction to novel objects in wild type, Wistar, Sprague-Dawley and Brown Norway rats

Background

The purpose of the present study was to analyse species-specific forms of behaviour (digging and swimming) and response to novelty in laboratory rats and their wild type counterparts at a very early stage of laboratorization. Three behavioural phenomena were taken into account: burrowing, spontaneous swimming, and neophobic behaviour.

Principal Findings

Wild-type rats and three strains of laboratory rats were involved in experiments: Warsaw-Wild-Captive-Pisula-Stryjek (WWCPS), Wistar, Sprague-Dawley, and Brown Norway rats were compared in spontaneous swimming test, while WWCPS and Wistar rats were studied in burrowing and neophobia experiments. Wild rats were found to be faster at building tunnels than Wistar rats and at constructing more complex burrow systems. The experiment on neophobia showed that Wistar rats exhibited less neophobic responses and were more often trapped. WWCPS rats showed highly neophobic behaviour and were rarely trapped in this experiment. The experiment on swimming showed that WWCPS rats showed more complex water tank related activity than their laboratory counterparts. They swam and explored under surface environment.

Conclusions

The three experiments showed profound behavioural differences in quasi-natural forms of behaviour between wild type rats (WWCPS) and three laboratory strains frequently used in behavioural studies.     

Genetic control of estrogen action in the rat: mapping of QTLs that impact pituitary lactotroph hyperplasia in a BN × ACI intercross

Estrogens are important regulators of growth and development and contribute to the etiology of several types of cancer. Different inbred rat strains exhibit marked, cell-type-specific differences in responsiveness to estrogens as well as differences in susceptibility to estrogen-induced tumorigenesis. Regulation of pituitary lactotroph homeostasis is one estrogen-regulated response that differs dramatically between different inbred rat strains. In this article we demonstrate that the growth response of the anterior pituitary gland of female ACI rats to 17β-estradiol (E2) markedly exceeds that of identically treated female Brown Norway (BN) rats. We further demonstrate that pituitary mass, a surrogate indicator of absolute lactotroph number, behaves as a quantitative trait in E2-treated F₂ progeny generated in a genetic cross originating with BN females and ACI males. Composite interval mapping analyses of the (BN×ACI)F₂ population revealed quantitative trait loci (QTLs) that exert significant effects on E2-induced pituitary growth on rat chromosome 4 (RNO4) (Ept5) and RNO7 (Ept7). Continuous treatment with E2 rapidly induces mammary cancer in female ACI rats but not BN rats, and QTLs that impact susceptibility to E2-induced mammary cancer in the (BN×ACI)F₂ population described here have been mapped to RNO3 (Emca5), RNO4 (Emca6), RNO5 (Emca8), RNO6 (Emca7), and RNO7 (Emca4). Ept5 and Emca6 map to distinct regions of RNO4. However, Ept7 and Emca4 map to the same region of RNO7. No correlation between pituitary mass and mammary cancer number at necropsy was observed within the (BN×ACI)F₂ population. This observation, together with the QTL mapping data, indicate that with the exception of the Ept7/Emca4 locus on RNO7, the genetic determinants of E2-induced pituitary growth differ from the genetic determinants of susceptibility to E2-induced mammary cancer.     

Response of thirsty rats to absence of water: Frustration,disinhibition or compensation?

Water-deprived rats were allowed to drink in an experimental chamber during a series of training trials, and were then presented with an empty water spout during a single test trial. Their initial response to absence of water was to bite, sniff and paw at the empty spout (spout-directed behaviour), after which they engaged in activities such as eating, grooming and exploration of the chamber. By comparison with control animals that had no expectation of water, experimental rats performed these latter activities in a bizarre ‘speeded-up’ manner, which was quantifiable as an increase in local rate. Both the occurrence of spout-directed behaviour and the increase in local rate of other activities were enhanced by increasing the prior level of water deprivation and by offering a more palatable fluid during training. The results are discussed in relation to current models of behavioural switching.     

Changes in innate forms of motor behavior in rats during long-term hypokinesia

The structure of motor behaviour in response to environmental tests--"open field" and "with-a-partner" situation (according to P. V. Simonov)--was studied in rats after 1, 3 and 6 weeks of hypokinesis, (to K. Hecht), as compared to the control. In control animals a relatively low level of orienting-investigation reactions and of grooming was observed as well as a low alimentary activity, which is considered as a manifestation of innate passive defensive reactions in the above situations. Disappearance of these reactions in the course of hypokinesia has two stages: predominant increase in orienting-investigating reactions (1-3 weeks); their subsequent decrease and an increase in the grooming (after 6 weeks). The relation between innate and conditioned behavioural changes is discussed as well as stability of alimentary behaviour in rats.     

Features of the behavior of the rat with hereditarily determined arterial hypertension

The behaviour of spontaneously hypertensive rats (SHR strain), rats with inherited stress induced arterial hypertension (ISIAH, a new developed strain), and of their normotensive Wistar progenitors was studied. The open-field arena and a device for measuring the total activity in the home cage were used in the behavioural studies. The SHR were much more active in the open--field and home cage tests than the Wistar and ISIAH rats. The basal locomotor activity of the ISIAH strain was lower than that of the Wistar rats, but the ISIAH strain had an index of behavioural reactivity 2.7 fold higher than the Wistar or SHR strains. These behavioural characteristics corresponded to the hypertension patterns of the strains compared. Enhanced spontaneous locomotion of the SHR rats was associated with spontaneous increase in arterial blood pressure. The ISIAH rats showed low spontaneous locomotor activity, but high behavioural and blood pressure reactivity under conditions of mild emotional stress.     

The evolutionary implications of qualitative variation in the grooming behaviour of the Hymenoptera (Insecta)

The grooming behaviour of 115 species of Hymenoptera (including castes and sexes) was recorded on an event recorder. Twenty-eight primary and secondary grooming movements are described, and eighteen are illustrated. An analysis of the grooming behaviour revealed consistent qualitative behavioural differences between virtually every super-family. The pattern of these differences parallels an existing hymenopteran phylogeny. Limited data suggest possible qualitative differences at the family and sub-family level in some instances. Possible use of grooming data in phylogenetic interpretatons is suggested.     

The antinociceptive efficacy of buprenorphine administered through the drinking water of rats

Postoperative pain management in laboratory animals is important for animal welfare and required under law in many countries. Frequent injection of analgesics to rodents after surgery is stressful for the animals and labour-intensive for animal care personnel. An alternative dosing scheme such as administration of analgesics in the drinking water would be desirable. However, the efficacy of a chronic oral analgesic treatment via this route has not yet been documented. This study investigated the antinociceptive efficacy of buprenorphine administered ad libitum via the drinking water of laboratory rats. The antinociceptive efficacy of buprenorphine in drinking water was compared with repeated subcutaneous injections. A comparison was also made between buprenorphine in drinking water and the combination of one single subcutaneous injection of buprenorphine followed by buprenorphine in drinking water. Antinociception was assessed by use of an analgesiometric model measuring the rats' latency time to withdrawal from a noxious heat stimulus applied to the plantar surface of the paw. Results revealed that buprenorphine in drinking water (0.056 mg/mL) induced significant increases in paw withdrawal latency times during a three-day period of administration with a maximal effect at 39 h after the start of buprenorphine administration. One single injection of buprenorphine (0.1 mg/kg s.c.) followed by buprenorphine in the drinking water (0.056 mg/mL) induced an earlier onset of antinociception than buprenorphine in drinking water alone. In contrast, buprenorphine (0.1 mg/kg s.c.) injected every 8 h over a period of three days did not result in significant increases in paw withdrawal latency times. In conclusion, our results suggest that one single subcutaneous injection of buprenorphine followed by buprenorphine in drinking water may be a viable treatment option for the relief of pain in laboratory rats, but at the doses used in this study in pain-free rats it was associated with a decrease in water intake and some behavioural changes.     

Evaluation of the general suitability of the rat for the micronucleus assay: the effect of cyclophosphamide in 14 strains.

To evaluate the general suitability of the rat for the micronucleus assay, we conducted the assay in males of 14 different strains, 13 inbred (ACI, BN, BUF, COP, DRH, F344, IS, LEW, RCS, SHR, WAG, WKYO, WTC) and 1 outbred (SD), using cyclophosphamide as the test chemical. Cyclophosphamide at 0 (vehicle), 5, 10, or 20mg/kg per day was administered orally twice, 24h apart, to five rats per dosage group. Bone marrow and peripheral blood were collected 24h after the second treatment.All 14 strains showed a positive response to cyclophosphamide, with slight differences in sensitivity. We concluded that the rat is suitable for the micronucleus assay regardless of strain.     

Behaviour of the Common Moorhen in Rio Grande do Sul, Brazil

This study presents data on behavioural acts performed by the Common Moorhen Gallinula chloropus in southern Brazil, and compares these with the behaviours previously reported for other populations. Focal observations of individuals were conducted in the municipality of Santa Maria, in the central region of the state of Rio Grande do Sul, Brazil. The sampling was done in 2-hour sessions, between January and March of 2007. A total of 20 behavioural acts, grouped in seven categories, were identified and described: locomotion (N = 5 acts), grooming (N = 4), intra-specific behaviour (N = 2), inter-specific behaviour (N = 3), foraging (N = 2), reproduction (N = 2) and rest (N = 2). Among the observed behaviours were acts that are not described in the literature such as greeting of offspring and some feeding acts. Regarding the use of habitat, we observed that this species has a preference for water or aquatic macrophytes, which is contrary to other reports. In the analysis of behavioural daily variation, overall behavioural categories did not vary significantly throughout the day, whereas we observed a significant difference in the use of categories during the periods 11:00 am–1:00 pm, 1:00–3:00 pm and 5:00–7:00 pm. The contrasting data between studies indicate that the variation between habitats and ecological interactions may generate different selective pressures on the behaviour of G. chloropus.     

秦岭川金丝猴的昼间活动时间分配

1999年12月至2004年11月,通过野外直接跟踪观察,采用焦点取样和瞬时记录的方法,对秦岭川金丝猴西梁猴群的昼间活动时间分配进行了研究。将川金丝猴的昼间活动定义为摄食、休息、理毛、移动和其他行为5 种类型,结果发现：川金丝猴昼间活动中有两个摄食高峰,分别在每天的上午和下午,中午有一个较长时间的休息期。川金丝猴昼间的主要行为类型是摄食和休息（约占全部活动时间的62%）,各种行为依所占时间比例依次为：摄食（34.64 ± 1.70%）、休息（26.83 ± 1.69%）、理毛（11.58 ± 0.96%）、移动（18.93 ±1.08%）和其他行为（8.02 ± 0.82%）。在不同季节各种行为类型所占时间比例有明显的变化,表现为摄食和休息行为占昼间活动时间比例在夏（75.00%）、秋季（74.15%） 大于春（52.74%）、冬季（56.64%）;而移动、理毛和其他行为所占时间比例之和在夏（25.00%）、秋季（ 25.85%） 则明显小于冬（ 43.4%）、春季（47.3%）。不同季节间的活动时间分配经非参数检验（ Kruskal Wallis H test,N = 199）,理毛行为（ x² =11.438,df = 3,P < 0.05）有显著差异,摄食（x² = 20.469,df = 3,P < 0.01）、休息（x² = 27.235,df = 3,P < 0.01）、移动（x² = 24.110,df = 3,P < 0.01） 和其他行为（ x² = 60.810,df = 3,P < 0.01） 季节间差异非常显著;除摄食行为外,其余各行为类型的时间分配在不同性别-年龄组间也存在显著差异（P < 0.05）。     

Ventilatory phenotypes among four strains of adult rats.

Our purpose in this study was to identify different ventilatory phenotypes among four different strains of rats. We examined 114 rats from three in-house, inbred strains and one outbred strain: Brown Norway (BN; n = 26), Dahl salt-sensitive (n = 24), Fawn-hooded Hypertensive (FHH: n = 27), and outbred Sprague-Dawley rats (SD; n = 37). We measured eupneic (room air) breathing and the ventilatory responses to hypoxia (12% O(2)-88% N(2)), hypercapnia (7% CO(2)), and two levels of submaximal exercise. Primary strain differences were between BN and the other strains. BN rats had a relatively attenuated ventilatory response to CO(2) (P < 0.001), an accentuated ventilatory response to exercise (P < 0.05), and an accentuated ventilatory roll-off during hypoxia (P < 0.05). Ventilation during hypoxia was lower than other strains, but hyperventilation during hypoxia was equal to the other strains (P > 0.05), indicating that the metabolic rate during hypoxia decreased more in BN rats than in other strains. Another strain difference was in the frequency and timing components of augmented breaths, where FHH rats frequently differed from the other strains, and the BN rats had the longest expiratory time of the augmented breaths (probably secondary to the blunted CO(2) sensitivity). These strain differences not only provide insight into physiological mechanisms but also indicate traits (such as CO(2) sensitivity) that are genetically regulated. Finally, the data establish a foundation for physiological genomic studies aimed at elucidating the genetics of these ventilatory control mechanisms.     

Orbital sinus blood sampling in rats: effects upon selected behavioural variables

The question addressed was whether the behaviour of rats is changed after orbital sinus blood sampling while they are under diethyl-ether anaesthesia. Twelve rats were subjected to sham anaesthesia, diethyl-ether anaesthesia and anaesthesia plus orbital puncture according to a Latin square. After each treatment, the behaviour of the rats was automatically monitored using the so-called LABORAS method, which discriminates between grooming, locomotion and inactivity. Treatment ended, and behaviour monitoring began, when the light period changed over to the dark period. The various behaviours were quantified as relative duration and frequency. Anaesthesia versus sham anaesthesia reduced the relative duration of grooming during the first 5 h after treatment. Anaesthesia plus orbital puncture versus anaesthesia alone did not significantly influence grooming, but orbital puncture did reduce the relative duration and the frequency of locomotion during the entire 20 h period, which was mainly due to a decrease in the dark period. After orbital puncture, the animals were also less frequently inactive. It is concluded that orbital puncture has an effect on behaviour superimposed on that of diethyl-ether anaesthesia. This observation may contribute new arguments to the debate on the acceptability of the orbital puncture technique.     

Strain dependence of the airway response to dry-gas hyperpnea challenge in the rat

The aim of thestudy was to investigate strain dependence and mechanisms of airwayresponses to dry-gas hyperpnea challenge in the rat. We studiedresponses in a strain that is hyperresponsive to methacholine, Fischer344 (F-344); in two normoresponsive strains, Lewis and ACI; and in anatopic but normoresponsive strain, Brown Norway (BN). We examined theeffects of a neurokinin (NK) 1-receptor (CP-99994), anNK₂-receptor (SR-48968), and aleukotriene D₄(LTD₄)-receptor antagonist(pranlukast) on responses to hyperpnea challenge in BN rats. Theanimals were ventilated with a tidal volume of 8 ml/kg and a frequencyof 150 breaths/min with either a dry or humidified mixture of 5%CO₂-95%O₂ for 5 min for hyperpneachallenge, whereas responses to challenge were measured duringspontaneous breathing. Pulmonary resistance increased after dry-gaschallenge in BN and ACI but not in F-344 and Lewis rats. CP-99994,SR-48968, and pranlukast significantly attenuated the increase inpulmonary resistance after dry-gas challenge. There were no significant differences in responsiveness to airway challenge withLTD₄ among the BN, F-344 and ACIrats. We conclude that responses to dry-gas hyperpnea challenge arestrain dependent in rats and are mediated by NKs andLTD₄.