Medicinal Mascarene Aloes: An audit of their phytotherapeutic potential.

A phytochemical and biological investigation of the endemic Mascarene Aloes (Aloe spp.), including A. tormentorii (Marais) L.E.Newton & G.D.Rowley, A. purpurea Lam, A. macra Haw., A. lomatophylloides Balf.f and A. vera (synonym A. barbadensis Mill.), which are used in the traditional folk medicine of the Mascarene Islands, was initiated. Methanolic extracts of the Aloes under study were analysed using high resolution LC-UV-MS/MS and compounds belonging to the class of anthraquinones, anthrones, chromones and flavone C-glycosides were detected. The Mascarene Aloes could be distinguished from A. vera by the absence of 2″-O-feruloylaloesin and 7-O-methylaloeresin. GC-MS analysis of monosaccharides revealed the presence of arabinose, fucose, xylose, mannose and galactose in all the Mascarene Aloes and in A. vera. The crude extracts of all Aloes analysed displayed antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Only extracts of A. macra were active against P. aeruginosa and Klebsiella pneumoniae, while none of the Aloe extracts inhibited Propionibacterium acnes. A. macra displayed anti-tyrosinase activity, exhibiting 50% inhibition at 0.95mg/mL, and extracts of A. purpurea (Mauritius) and A. vera displayed activity in a wound healing-scratch assay. In vitro cytotoxicity screening of crude methanolic extracts of the Aloes, using the MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) showed that only A. purpurea (Réunion) elicited a modest toxic effect against HL60 cells, with a percentage toxicity of 8.2% (A. purpurea-Réunion) and none of the Aloe extracts elicited a toxic effect against MRC 5 fibroblast cells at a concentration of 0.1mg/mL. Mascarene Aloe species possess noteworthy pharmacological attributes associated with their rich phytochemical profiles.

Bio-prospecting endemic Mascarene Aloes for potential neuroprotectants.

The Mascarene Aloes are used in the traditional pharmacopoeia against various ailments including cutaneous diseases and as antispasmodics. Scientific evidence to support these claims is non-existent and mainly based on the scientific repute of A. vera. The antioxidant profile of methanolic leaf extracts of A. purpurea Lam., A. tormentorii (Marais) L. E. Newton & G. D. Rowley, A. lomatophylloides Balf. f., A. macra Haw. and A. vera (L.) Burm. f. was studied using the total antioxidant capacity, copper equivalent and superoxide dismutase assays. In vitro cytotoxicity was evaluated on CAD (Cath.-a-differentiated) neuronal cells by the methyl tetrazolium assay, and the neuroprotective profile was assessed using hydrogen peroxide-induced neurotoxicity with the CAD cells. The aloin and vitexin content were determined by high-performance liquid chromatography with diode-array detection. A. purpurea had the highest aloin content (546.6 nmol/g), while A. tormentorii had the highest vitexin content (67.3 nmol/g). A. macra (concentration <0.1 mg/mL) elicited a 10% cytotoxicity effect on CAD cells while other Mascarene Aloes were not cytotoxic. This study validates the antioxidant and neuroprotective potential of Mascarene Aloes focusing on their aloin and vitexin content that are also present in other reputed medicinal Aloes.

Slope climbing challenges, fear of heights, anxiety and time of the day.

When exposed to an unfamiliar open space, animals experience fear and attempt to find an escape route. Anxiety emerges when animals are confronted with a challenging obstacle to this fear motivated escape. High anxiety animals do not take risks; they avoid the challenge. The present experiments investigated this risk avoidant behavior in mice. In experiment 1, BALB/c, C57BL/6J and CD-1 mice were exposed to a large platform with downward inclined steep slopes attached on two opposite sides. The platform was elevated 75 and 100cm from the ground, in a standard (SPDS) and in a raised (RPDS) configuration, respectively. In experiment 2, the platform was elevated 75cm from the ground. Mice had to climb onto a stand at the top of upward inclined slopes (SPUS). In experiment 3, BALB/c mice were exposed to SPDS with steep or shallow slopes either in early morning or in late afternoon. In all 3 test configurations, mice spent more time in the areas adjacent to the slopes than in the areas adjacent to void, however only C57BL/6J and CD-1 crossed onto the slopes in SPDS, and crossed onto the stands in SPUS whereas BALB/c remained on the platform in SPDS and explored the slopes in SPUS. Elevation of the platform from the ground reduced the crossings onto the slopes in C57BL/6J and CD-1, and no differences were observed between BALB/c and C57BL/6J. BALB/c mice demonstrated no difference in anxiety when tested early morning or late afternoon; they crossed onto shallow slopes and avoided the steep one.

Pre-training in a radial arm maze abolished anxiety and impaired habituation in C57BL6/J mice treated with dizocilpine.

Familiarity can imply a reduction of fear and anxiety, which may render learning and memory performance insensitive to NMDA receptor antagonism. Our previous study indicates that MK-801 (dizocilpine), NMDA antagonist, increased anxiety and prevented the acquisition of a spatial memory task. Here, we examined whether MK-801 will produce anxiety in mice that were familiar with the test environment. Male C57BL/6J mice were exposed, one session a day for 7days, to a 3D maze, which consisted of nine arms attached to upward inclined bridges radiating from a nonagonal platform. In this maze, high anxiety mice avoid the arms in the first sessions. One group of mice received saline (SAL) while a second group received MK-801 (MKD1), both on day one. A third group received saline in the first 3 sessions, and MK 801 in subsequent sessions (MKD4). Saline and MK-801 (0.1mg/kg) were administered intraperitoneally 30min before the test. MKD4 mice demonstrated an increase in bridge and arm visits, and reached arm/bridge entries ratio close to 1 in session 5. SAL mice also crossed frequently onto the arms, and reached a comparable ratio, but this was achieved with a lower number of arm visits. MKD1 mice demonstrated a reduced number of arm visits in each session compared to SAL and MKD4 mice. Dizocilpine produced anxiety in mice treated from day 1 of the test, but not in those treated from day 4. It also impaired habituation in animals familiar with the test environment; it produced sustained non-habituating hyperactivity.

Altered GABAA α5 subunit expression in the hypothalamic paraventricular nucleus of hypertensive and pregnant rats.

A characteristic of both hypertension and pregnancy is increased sympathetic nerve activity. The level of sympathetic activation is determined, in part, by a tonic GABAergic inhibition arising from the hypothalamic paraventricular nucleus (PVN). In hypertension, decreases in GABAergic inhibition and increases in glutamatergic excitation within the PVN contribute to this sympatho-excitation. In late-term pregnancy however, the sympatho-excitation appears to be mediated by decreases in GABAergic inhibition only. This study examined whether changes in subunit expression for GABAA receptors in the PVN could provide a molecular basis for the sympatho-excitation characteristic of hypertension and pregnancy. Hypertension and pregnancy were accompanied by significant decrease in the GABAA receptor α5 subunit in the PVN. We suggest that decreases in the α5 subunit of the GABAA receptor may be important in mediating the sympatho-excitation observed in both hypertension and pregnancy.

Effects of chronic fluoxetine treatment on anxious behaviour of BALB/c mice in a 3-dimensional maze.

Here we used a 3-dimensional (3D) maze, a modification of the radial maze, to assess the effects of treatment for two weeks with a single daily dose of fluoxetine (20 mg/kg, i.p.) on anxiety in male BALB/c mice. We examined whether anxiolytic effects of fluoxetine can be detected over three daily test sessions. We examined also whether repeated handling associated with chronic treatment interferes with effects of fluoxetine on anxiety responses. The 3D maze comprises nine arms, each connected to an upward inclined bridge radiating from a central platform. In this maze, BALB/c mice cross frequently into the bridges but avoid the arms. This avoidance is used as an index of anxiety. Two separate groups received once a day either saline (SALCH, n = 8) or fluoxetine (FLUCH, n = 8) for 14 days, and up to 30 min before the test during the subsequent 3 days. A third group received saline (SALAC, n = 8) 30 min before the test, once a day for 3 days. SALAC mice did not cross into the arms, and continued this avoidance over 3 sessions. SALCH mice avoided the arms in session 1 whereas FLUCH mice did cross into the arms, and like SALCH mice, increased number of crossings into and time on the arms in subsequent sessions. Fluoxetine evidently had an anxiolytic effect but only in the first session. These results indicate that handling experience decreased fear and anxiety in the mice, which may have masked the anxiolytic effect of fluoxetine in the second and third test sessions.

Antagonism of histamine H4 receptors exacerbates clinical and pathological signs of experimental autoimmune encephalomyelitis.

BACKGROUND AND PURPOSE: The histamine H4 receptor has a primary role in inflammatory functions, making it an attractive target for the treatment of asthma and refractory inflammation. These observations suggested a facilitating action on autoimmune diseases. Here we have assessed the role of H4 receptors in experimental autoimmune encephalomyelitis (EAE) a model of multiple sclerosis (MS). EXPERIMENTAL APPROACH: We induced EAE with myelin oligodendrocyte glycoprotein (MOG35-55 ) in C57BL/6 female mice as a model of MS. The histamine H4 receptor antagonist 5-chloro-2-[(4-methylpiperazin-1-yl)carbonyl]-1H-indole (JNJ7777120) was injected i.p. daily starting at day 10 post-immunization (D10 p.i.). Disease severity was monitored by clinical and histopathological evaluation of inflammatory cells infiltrating into the spinal cord, anti-MOG35-55 antibody production, assay of T-cell proliferation by [(3) H]-thymidine incorporation, mononucleate cell phenotype by flow cytometry, cytokine production by elisa assay and transcription factor quantification of mRNA expression. KEY RESULTS: Treatment with JNJ7777120 exacerbated EAE, increased inflammation and demyelination in the spinal cord of EAE mice and increased IFN-γ expression in lymph nodes, whereas it suppressed IL-4 and IL-10, and augmented expression of the transcription factors Tbet, FOXP3 and IL-17 mRNA in lymphocytes. JNJ7777120 did not affect proliferation of anti-MOG35-55 T-cells, anti-MOG35-55 antibody production or mononucleate cell phenotype. CONCLUSIONS AND IMPLICATIONS: H4 receptor blockade was detrimental in EAE. Given the interest in the development of H4 receptor antagonists as anti-inflammatory compounds, it is important to understand the role of H4 receptors in immune diseases to anticipate clinical benefits and also predict possible detrimental effects.

Non-invasive infra-red therapy (1072 nm) reduces ß-amyloid protein levels in the brain of an Alzheimer's disease mouse model, TASTPM.

BACKGROUND: Alzheimer's disease (AD) is the most prevalent neurodegenerative disease and common cause of dementias in the Western world. This study investigated the expression profile of heat-shock proteins (HSPs) involved in maintaining healthy neurons in the TASTPM AD mouse model, and whether chronic treatment with 1072 nm infra-red (IR1072) modified the expression profiles of HSPs and amyloidopathy in female TASTPM mice. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative immunoblotting and immunohistochemistry were used to examine the expression of proteins such as HSPs, phosphorylated tau (tau-P), amyloid precursor protein (APP), ß-amyloid1-40 (Aß), and Aß1-42. TASTPM mice at 3, 7 and 12 months were investigated as well as female TASTPM mice which had undergone a chronic, 5 month, IR1072 treatment. During the first 12 months of age, a critical period of AD progression, reduced HSP40 and HSP105 were observed. αB-crystallin, Aß1-42 and tau-P increased over this period, particularly between 3 and 7 months. Chronic IR1072 treatment of female TASTPM mice elicited significant increases in HSP60, 70 and 105 and phosphorylated-HSP27 (P-HSP27) (50-139%), together with a concomitant profound decrease in αB-crystallin, APP, tau-P, Aß1-40 and Aß1-42 (43-81%) protein levels at 7 months of age. Furthermore, IR1072 treatment elicited a modest, but significant, reduction in Aß1-42 plaques in the cerebral cortex. CONCLUSIONS/SIGNIFICANT FINDINGS: IR1072 treatment provides a novel non-invasive and safe way to upregulate a panel of stress response proteins in the brain, known to both reduce protein aggregation and neuronal apoptosis. This approach recently entered clinical trials for AD in the USA, and may provide a novel disease modifying therapy for a range of neuropathologies.

Overexpression of histamine H4 receptors in the kidney of diabetic rat.

OBJECTIVE AND DESIGN: The renal expression of H1 and H2 receptors has previously been demonstrated, while that of the H4 receptor has been poorly investigated, and thus the aim of this research was to investigate the expression of the H4 receptor in the kidney of diabetic rats. MATERIAL OR SUBJECTS: 24 8-week-old male Wistar rats. TREATMENT: Diabetes was induced in 12 rats by a single intravenous injection of streptozotocin, and animals were killed 6 weeks later. METHODS: Kidneys were collected and processed for quantitative PCR or immunohistochemical analyses. To ascertain the renal topology of the H4 receptor, colocalization experiments were performed with a series of markers. RESULTS: H4 receptor is expressed in healthy rats, although at a very low level, and is strongly upregulated in diabetic animals. Immunohistochemical analysis revealed the highest immune-positivity in the medulla. Colocalization experiments revealed a close overlap in expression topology of the H4 receptor and both Tamm-Horsfall glycoprotein and aquaporin 1 was observed. CONCLUSIONS: The results demonstrate, for the first time, that the H4 receptor is expressed in the kidney mainly by resident renal cells of the loop of Henlé and that this receptor is significantly overexpressed in diabetic animals, thus suggesting a possible role in the pathogenesis of diabetes-associated renal disease.

Brief report: first identification of H4 histamine receptor in healthy salivary glands and in focal sialadenitis in Sjögren's syndrome.

OBJECTIVE: The conventional H(1) and H(2) histamine receptors have >10,000-fold lower avidity for histamine than H(4) histamine receptor, which has been implicated in autoimmune diseases. This study was undertaken to compare H(4) histamine receptor levels in the salivary glands (SGs) of healthy controls with those in the SGs of patients with primary Sjögren's syndrome (SS). METHODS: H(4) histamine receptor messenger RNA (mRNA) was analyzed using real-time quantitative polymerase chain reaction, and the receptor protein was examined using immunostaining. Effects of the H(4) histamine receptor agonist ST-1006 on cytokine synthesis by human SG (HSG) cells were analyzed using xMAP technology and enzyme-linked immunosorbent assay. RESULTS: Healthy SGs contained H(4) histamine receptor mRNA. The receptor protein was localized to the acinar and ductal epithelial cells. H(4) histamine receptor agonist stimulated HSG cells to produce the cytokines interleukin-8 and vascular endothelial growth factor. SS patients had low H(4) histamine receptor levels. CONCLUSION: H(1) and H(2) histamine receptor antagonists are not effective in the treatment of autoimmune diseases. However, such antagonists do not affect the newly discovered H(4) histamine receptor. Dendritic cells and lymphocytes are nonprofessional histamine-producing cells, which produce histamine at 100-1,000-fold lower rates than mast cells do. Saliva contains only 0.31-12.4 ng/ml histamine, which is too low to stimulate H(1) or H(2) histamine receptor, but stimulates H(4) histamine receptor half maximally. Our findings show that H(4) histamine receptor is strongly expressed in tubuloacinar SG cells, which emphasizes the role of these cells in the pathogenesis of SS.

MK-801 increases the baseline level of anxiety in mice introduced to a spatial memory task without prior habituation.

C57BL/6J mice were introduced to a nine arm radial maze without prior habituation and trained in the acquisition of a working memory task in 16 sessions, one session per day. In this maze mice need to climb onto an upward inclined bridge in order to reach and cross onto an arm. They received in each session an i.p. injection of MK-801 (0.1 mg/kg) 30 min before training or immediately after training. MK-801 pre-treated mice made significantly more entries onto the bridges, fewer entries onto the arms and took significantly longer time to make a first arm visit compared to saline and MK-801 post-treated mice during the first 3 session blocks (4 sessions per block). These results indicate that MK-801 induced anxiety which was extended throughout the first 3 session blocks. MK-801 pre-treated mice made also significantly more errors and required more sessions to reach the criterion compared to saline and MK-801 post-treated mice. Administration of MK-801 after training did not affect the acquisition of the task. The present results indicate that MK-801 pre-treatment impaired the acquisition of a spatial task and this can be accounted for by its effect on the baseline level of anxiety which was elevated. The introduction of mice to the acquisition of the task without prior habituation demonstrates that a drug treatment can affect learning and memory by increasing and/or prolonging anxiety. Such effect may be confounded with learning and memory performance and not detected with pre-habituation training procedures, particularly when the number of sessions is determined a-priori.

Tolerance, sensitization and dependence to diazepam in Balb/c mice exposed to a novel open space anxiety test.

Balb/c mice were exposed to an elevated platform that is extended on two opposite sides with lowered steep slopes. They were tested for 12min per session in 6 successive days. They received i.p. administration of either saline or one dose of diazepam (DZP 0.5, 1, 3mg/kg) in sessions 1-3, and saline in sessions 4 and 5. All groups of mice received a single dose of DZP (1mg/kg) in session 6. DZP produced inverted U-shaped dose-responses on the number of entries into different areas of the apparatus, with a peak in mean response at 1mg/kg whereas its effect on the duration of entries was mostly comparable between the 3 doses. It increased the number of crossings on the surface of the platform and facilitated entries onto the slopes. DZP-treated mice crossed frequently onto and spent longer time on the slopes in sessions 1-3 whereas saline-treated mice remained on the platform in sessions 1-6. Withdrawal of DZP in sessions 4-5 increased the latency of first entry and decreased the number and duration of entries onto the slopes which was reversed with the administration of 1mg/kg of DZP in the next session. This ON-OFF the drug may be due to the half-life of DZP which is very short in mice and rats ( approximately 0.88h). It also indicates that DZP-treated mice did not benefit from previous experience of entries onto the slopes which suggests a possible "state-dependent" effect. Administration of DZP after repeated exposures to the test did not facilitate entries onto the slopes but instead increased significantly the number of crossings on the surface of the platform; this increase was much higher than that observed in mice initially treated with DZP and exposed to the test. There is no evidence of habituation in saline-treated mice: the number of crossings on the platform was comparable between the first 5 sessions of the test. These results demonstrate that repeated exposures to the same anxiogenic environment resulted in avoidance responses developing tolerance and approach responses developing sensitization. They suggest that tolerance and sensitization are two opposite sides of the habituation process to the same stimulus and may account for the maintained state of anxiety.

Anxiety responses in Balb/c, c57 and CD-1 mice exposed to a novel open space test.

In the present report we describe the behavior of two albinos (BALB/c and CD-1) and one pigmented (c57/BL6) strains of mice exposed to a novel open space anxiety test in a single 12 min session. The test is based on exposure of mice to an unfamiliar elevated platform which is extended on two opposite sides with steep slopes presented downward or upward. In the first experiment, the behavior of mice was examined on the elevated platform at two different heights (75 and 100 cm) with downward slopes. In the second experiment, we examined the behavior of mice on the platform at the lowest height (75 cm) but with upward slopes which lead to a stand. In the third experiment, we examined the behavior of Balb/c mice on the platform at the lowest height (75 cm) with downward slopes, and a hub enclosure providing a protected space located in the centre of the platform. The least anxious strain of mice was expected to take risks and cross onto the slopes (experiments 1 and 3) and onto the stands (experiment 2). The results of experiment 1 show that Balb/c mice did not cross onto the slope, and CD-1 mice made more crossings into and spent more time on the slopes than c57 mice. The increase in the heights of the platform reduced the number of crossings on the platform in all three strains of mice, and decreased the time spent on the platform before first entry onto a slope in c57 and in CD-1 mice. It also decreased the number of entries and duration of entries onto the slopes in CD-1 mice. In experiment 2, Balb/c mice did cross onto the upward slopes but significantly less than c57 and CD-1 mice but they did not cross onto the stands attached to the end of the slopes. CD-1 mice made more entries onto and spent more time on the stands than c57 mice. In the third experiment, Balb/c and c57 mice spent most of their time inside a protective space (cylinder) placed in the centre of the platform demonstrating strong avoidance responses of the outer area of the platform, and only three c57 mice crossed onto the slopes for a very brief duration in one or two entries. In all three experiments, mice entered more frequently and spent more time in the outer areas than in the inner areas of the platform, particularly in the areas adjacent to slopes than in the areas adjacent to a void space. CD-1 mice appears the least anxious taking more risks by venturing onto the slopes and onto the stands while Balb/c appears the most anxious spending a large amount of time in the areas adjacent to the slopes. The different configurations of the test apparatus (experiments 1 and 2) seem to provide different incentives for the drive to explore and escape which may account for differences in anxiety responses whereas the presence of a protective space (experiment 3) appears to encourage avoidance responses.

Distinguishing anxiolysis and hyperactivity in an open space behavioral test.

This report describes the emotional responses of mice exposed to an unfamiliar elevated platform that is extended on two opposite sides by downward lowered steep slopes. Balb/c mice were exposed to the test for 12 min per session in 3 successive days. They received i.p. administration of diazepam (0, 0.5, 1 and 3 mg/kg) or amphetamine (0, 1, 2.5, 5 and 10 mg/kg) 30 min prior to test sessions. Separate groups of Balb/c mice were used for each dose of the drugs. Both drugs increased the number of crossings on the platform, indicating increased motor activity, and the effects were dose-dependent. Diazepam also significantly increased the number and duration of entries onto the slopes indicating an anxiolytic effect, whereas none of the saline or amphetamine-treated mice adventured onto the slopes. Amphetamine and diazepam produced an inverted U-shaped dose-response effect on different parameters of the test and demonstrate that the drug concentration which elicited a peak in mean number of entries is different from the drug concentration which elicited a peak in mean duration of entries. This study demonstrates the sensitivity and discriminatory power of an open space anxiety test for future pharmacological studies.

Immunological identification of the mouse H4 histamine receptor on spinal cord motor neurons using a novel anti-mouse H4R antibody.

In this report, we describe the development of the first anti-mouse H(4) receptor antibody. Utilising this new immunological probe, new evidence is provided for oligomeric mH(4)Rs and the presence of H(4) receptors on a subpopulation of murine motor neurons. This expands the cell types where the H(4) receptor is expressed in the mammalian CNS.

The histamine H4 receptor is functionally expressed on neurons in the mammalian CNS.

BACKGROUND AND PURPOSE: The histamine H4 receptor is the most recently identified of the G protein-coupled histamine receptor family and binds several neuroactive drugs, including amitriptyline and clozapine. So far, H4 receptors have been found only on haematopoietic cells, highlighting its importance in inflammatory conditions. Here we investigated the possibility that H4 receptors may be expressed in both the human and mouse CNS. METHODS: Immunological and pharmacological studies were performed using a novel anti-H4 receptor antibody in both human and mouse brains, and electrophysiological techniques in the mouse brain respectively. Pharmacological tools, selective for the H4 receptor and patch clamp electrophysiology, were utilized to confirm functional properties of the H4 receptor in layer IV of the mouse somatosensory cortex. RESULTS: Histamine H4 receptors were prominently expressed in distinct deep laminae, particularly layer VI, in the human cortex, and mouse thalamus, hippocampal CA4 stratum lucidum and layer IV of the cerebral cortex. In layer IV of the mouse somatosensory cortex, the H4 receptor agonist 4-methyl histamine (20 micromol x L(-1)) directly hyperpolarized neurons, an effect that was blocked by the selective H4 receptor antagonist JNJ 10191584, and promoted outwardly rectifying currents in these cells. Monosynaptic thalamocortical CNQX-sensitive excitatory postsynaptic potentials were not altered by 4-methyl histamine (20 micromol x L(-1)) suggesting that H4 receptors did not act as hetero-receptors on thalamocortical glutamatergic terminals. CONCLUSIONS AND IMPLICATIONS: This is the first demonstration that histamine H4 receptors are functionally expressed on neurons, which has major implications for the therapeutic potential of these receptors in neurology and psychiatry.

A new model for the study of high-K(+)-induced preconditioning in cultured neurones: role of N-methyl-d-aspartate and alpha7-nicotinic acetylcholine receptors.

Spreading depression (SD), whether elicited by local application of high K(+) medium to the cortical surface or by other stimuli, can increase the brain's tolerance to a subsequent, severe ischaemic insult in vivo, a phenomenon termed preconditioning. Herein, we have developed and validated a robust in vitro protocol for high-K(+)-preconditioning of cultured neurones. This new model is especially appropriate to unravel the molecular mechanisms underlying neuronal preconditioning and subsequent ischaemic tolerance. With this new, optimised preparation, preconditioning was found to be dependent upon culture day in vitro, cell density, K(+) concentration and duration of treatment. Finally, preconditioning was shown to be dependent upon N-methyl-d-aspartate (NMDA), CAM-kinase II signalling and alpha7-nicotinic (alpha7 nACh) receptor function, which is analogous to in vivo preconditioning induced by various stimuli.

Do rats really express neophobia towards novel objects? Experimental evidence from exposure to novelty and to an object recognition task in an open space and an enclosed space.

Three set of experiments were performed in an enclosed space (open-field) and in an open space (elevated platform). The surface of the open-field and the elevated platform were divided in nine equal squares. Rats were exposed (without previous habituation) in a unique session (experiment 1) or three consecutive sessions (experiment 2) either to an open-field (enclosed space) or to an elevated platform (open space) with and without an object on the centre of the field. In experiment 3, rats were exposed (without previous habituation) either to an enclosed or an open space on five consecutive sessions, one session a day. They were tested in an object recognition test in sessions 1, 3 and 5. In sessions 2 and 4, no objects were present. In experiment 1, we recorded the latency, frequency and duration of entries into different areas of the field. In experiment 3, we recorded the latency, frequency and duration of contacts with objects in addition to entries into different areas of the field. The first experiment demonstrates that rats exposed for the first time to an enclosed or an open space do not express neophobia toward novel objects in the absence of walls that surround an open-field. They crossed frequently into and spent more time in areas occupied with an object than in unoccupied areas. After two sessions of habituation to an empty open space or an empty enclosed space, the latency of first approach to a novel object is reduced while the frequency and duration of approaches are increased. The third experiment on object recognition confirmed that rats do not avoid novel objects; they made frequent visit and spent more time in the corner of the field occupied with an object than in empty corners. Recording of crossings provided detailed information about the patterns of exploratory behavior of rats but failed to reveal discrimination between novel and familiar objects which was evident in both open and enclosed space with recording of contacts with objects on the fifth exposure.