Spatial learning and memory in male mice with altered growth hormone action

Growth hormone (GH) has a significant influence on cognitive performance in humans and other mammals. To understand the influence of altered GH action on cognition, we assessed spatial learning and memory using a Barnes maze (BM) comparing twelve-month old, male, bovine GH (bGH) and GH receptor antagonist (GHA) transgenic mice and their corresponding wild type (WT) littermates. During the acquisition training period in the BM, bGH mice showed increased latency, traveled longer path lengths and made more errors to reach the target than WT mice, indicating significantly poorer learning. Short-term memory (STM) and long-term memory (LTM) trials showed significantly suppressed memory retention in bGH mice when compared to the WT group. Conversely, GHA mice showed significantly better learning parameters (latency, path length and errors) and increased use of an efficient search strategy than WT mice. Our study indicates a negative impact of GH excess and a beneficial effect of the inhibition of GH action on spatial learning and memory and, therefore, cognitive performance in male mice. Further research to elucidate GH's role in brain function will facilitate identifying therapeutic applications of GH or GHA for neuropathological and neurodegenerative conditions.     

Maze-escape learning and memory during rat ontogeny

With the method of a simple automatic rectangular maze the development of escape reaction was studied in albino rats aged 3 weeks to 3 months as well as memory retrieval within 24 h and then after one month. Learning of escape improves sharply between the 3rd and 4th postnatal week, and then, with the exception of week 8, remains approximately on the same level. Memory retrieval up to 24 h is not significantly influenced by age or sex, but is best immediately after learning interval of 10 min and the poorest after 24 h, except the age of 5 weeks, where it is best in 3 and 6 h after learning. One-month retention of maze escape improves during development except in animals trained at the age of 6 weeks in which the level is lower than in all other investigated groups. One-month memory retrieval in animals taught at the age of 4 weeks is influenced by the time of the first retention test with the best values in animals tested for the first time after 3 h and poorest in animals tested after 24 h after learning. Animals trained at 5 weeks had a marginally better result after the one-month retention test in females, perhaps due to the influence of sexual maturation on memory traces. This was not observed in the other age groups. Attention is drawn to the different development of learning and memory of various types of conditioned reactions stemming from the difference in their complexity.     

The lens protein alpha-B-crystallin of the blind subterranean mole-rat: high homology with sighted mammals

Blind subterranean mole rats, Spalax ehrenbergi, retain a subcutaneous, degenerated eye, which is visually non-functional but which does function in circadian entrainment. Crystallins, members of the small heat shock protein family, constitute approximately 90% of the water-soluble proteins of the transparent eye lens and are crucial for its optical properties, but they are also expressed in other tissues. In our attempt to understand the role of the eye in the blind mole-rat, we now describe the cloning, sequencing, and expression of the cDNA of alpha-B-Crystallin from two species of Spalax (S. galili and S. Judaei, with diploid chromosome numbers 2n=52 and 60, respectively). Spalax alpha- B-Crystallin is highly conserved. It is expressed in many tissues of Spalax, among them Spalax eye. The sequence of the cDNA of alpha-B-Crystallin in the eye and in the heart of Spalax is identical. Further studies are essential to clarify the role of this gene in the lens of an atrophied eye of a visually blind mammal.     

Encephalization of Bathyergidae and comparison of brain structure volumes between the Zambian mole-rat Fukomys anselli and the giant mole-rat Fukomys mechowii

Encephalization indices were calculated for Fukomys anselli and Fukomys mechowii by using interspecific allometric lines of Tenrecinae (recent Eutheria with the smallest brains) and average Rodentia to compare brain sizes independent of body size influence. These were contrasted with corresponding indices of other Bathyergidae and additionally with other rodents. The Bathyergidae species had indices within the variation of some Cricetidae and Muridae and thus do not differ in encephalization. F. anselli, however, had a clearly higher encephalization index than the sister species F. mechowii. The sizes of diverse structures were measured in the brains of these two species by help of the serial section method. No differences were found in relative composition. The lower encephalization of F. mechowii is discussed as a special phenomenon of gigantism during phylogenetic radiation which similarly was documented for other forms.     

Reproductive features of the Russian vole in laboratory breeding.

Reproductive features of newly bred Russian voles (Microtus rossiaemeridionalis) as a laboratory animal were studied. This species is a copulatory ovulator, and most couples copulated 6 to 16 h after pairing. The gestation period varied from 18 to 22 days (mean +/- SD: 20.6 +/- 0.9, n = 72), and the average litter size was 4.6 +/- 1.9 (n = 125). Compared with the litter size at the first parturition (3.6 +/- 1.6, n = 72), the litter size in the subsequent parturitions increased to 5.9 +/- 1.4 (n = 53). The animals exhibited postpartum estrus, and repeated pregnancy accompanied with suckling pups and parturition continuously in the laboratory condition unlike other vole species. In view of their complex stomach and good proliferation, the Russian voles were evaluated as a good laboratory animal, especially as a model animal for ruminant studies.     

Spatial reference learning deficits in absence of dysfunctional working memory in the TgF344-AD rat model of Alzheimer's disease

Alzheimer's disease (AD) is characterized by cognitive disorders and alterations of behavioral traits such as anhedonia and anxiety. Contribution of nonphysiological forms of amyloid and tau peptides to the onset of neurological dysfunctions remains unclear because most preclinical models only present one of those pathological AD-related biomarkers. A more recently developed model, the TgF344-AD rat has the advantage of overexpressing amyloid and naturally developing tauopathy, thus making it close to human familial forms of AD. We showed the presence of a learning dysfunction in a reference memory test, without spatial working memory impairment but with an increase in anxiety levels and a decrease in motivation to participate in the test. In the sucrose preference test, TgF344-AD rats did not show signs of anhedonia but did not increase the volume of liquid consumed when the water was replaced by sucrose solution. These behavioral phenomena were observed at an age when tau accumulation are absent, and where amyloid deposits are predominant in the hippocampus and the entorhinal cortex. Within the hippocampus itself, amyloid accumulation is heterogenous between the subiculum, the dorsal hippocampus and the ventral hippocampus. Thus, our data demonstrated heterogeneity in the appearance of various behavioral and neurochemical markers in the TgF344-AD rat. This multivariate analysis will therefore make it possible to define the stage of the pathology, to measure its evolution and the effects of future therapeutic treatments.     

Spatial learning and memory retention in the grey bamboo shark (Chiloscyllium griseum)

Like other vertebrates, fish can learn to solve a wide variety of tasks; however, many of their cognitive abilities, particularly in cartilaginous fishes, still remain unknown. This study investigated memory retention capabilities of spatial learning tasks in the grey bamboo shark (Chiloscyllium griseum, n = 8) after behavioural training in a two-choice experiment (T-maze). Sharks learned to obtain food at a goal location, either by performing a specific turn response or by using a variety of external landmarks for guidance. Following successful training, sharks were subjected to several training breaks ranging from one to six weeks, after which they were tested again for memory retention capabilities. Sharks successfully mastered the spatial tasks after 5–21 sessions and retained this knowledge in the absence of reinforcement for a period of up to six weeks. This is the first study to show that sharks, like stingrays and other vertebrates, can solve spatial tasks and retain spatial knowledge for an extended period of time, possibly aiding them in activities such as food retrieval, predator avoidance, mate choice and habitat selection.     

Effect of dalargin on the processes of memory and learning in the rat

In experiments on rats, the influence was studied of dalargin on the elaboration and preservation of various homogeneous and heterogeneous conditioned reflexes (CRs) elaborated in single and multiple pairings. The effect of dalargin on the processes of learning and memory was compared with the action of the peptide on the activity of hypothalamic neurones. Administration of dalargin delayed the elaboration of maze defensive CRs and practically did not affect the elaboration of two-way avoidance. The preservation of CR also deteriorated under the influence of dalargin. Administration of dalargin 10 min before the CRs testing did not prevent their reproduction. When using CRs elaborated in a single pairing, dalargin disturbed the preservation of the drinking CR and improved that of passive avoidance CR. Dalargin in this dose affected the emotional state of animals in the open field and did not significantly affect their motor activity. Dalargin suppressed impulse activity in 17 out of 22 tested neurones of the lateral hypothalamus, with maximum effect in 20-50 min after its administration. The obtained data show that the character of dalargin action on the elaboration of CR and mainly on its consolidation, depends on the character of the elaborated CR and is probably due to great extent to the effect of the peptide on the brain emotional mechanisms.     

Short-term social memory in the laboratory rat: its susceptibility to disturbance

Adult rats presented with a juvenile conspecific for 5 min on two occasions, separated by a 15-min inter-exposure interval (IEI), investigated the reintroduced juvenile significantly less in the second encounter. It is suggested that this was because the adult rats remembered the identity of the juvenile, because when a novel juvenile was introduced for the second encounter, no such reduction in investigation was observed. When the rats were either handled, placed in a smaller, novel, cage, or introduced to a new juvenile midway through the IEI, investigation of the reintroduced juvenile did not decrease. This indicated that memory of that juvenile had been disrupted. However, a simple change of cage during the IEI had no disruptive effect on memory. These results suggest that routine husbandry procedures can disrupt short-term social memory, which may lead to an increase in aggression due to a failure of recognition. This has implications for the welfare of captive social animals.     

Detours by the blind mole-rat follow assessment of location and physical properties of underground obstacles

Orientation by an animal inhabiting an underground environment must be extremely efficient if it is to contend effectively with the high energetic costs of excavating soil for a tunnel system. We examined, in the field, the ability of a fossorial rodent, the blind mole-rat, Spalax ehrenbergi, to detour different types of obstacles blocking its tunnel and rejoin the disconnected tunnel section. To create obstacles, we dug ditches, which we either left open or filled with stone or wood. Most (77%) mole-rats reconnected the two parts of their tunnel and accurately returned to their orginal path by digging a parallel bypass tunnel around the obstacle at a distance of 10-20 cm from the open ditch boundaries or 3-8 cm from the filled ditch boundaries. When the ditch was placed asymmetrically across the tunnel, the mole-rats detoured around the shorter side. These findings demonstrate that mole-rats seem to be able to assess the nature of an obstacle ahead and their own distance from the obstacle boundaries, as well as the relative location of the far section of disconnected tunnel. We suggest that mole-rats mainly use reverberating self-produced seismic vibrations as a mechanism to determine the size, nature and location of the obstacle, as well as internal self-generated references to determine their location relative to the disconnected tunnel section. Copyright 2003 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.      

Spatial learning and memory deficits after whole-brain irradiation are associated with changes in NMDA receptor subunits in the hippocampus

Whole-brain irradiation is used for the treatment of brain tumors, but can it also induce neural changes, with progressive dementia occurring in 20-50% of long-term survivors. The present study investigated whether 45 Gy of whole-brain irradiation delivered to 12-month-old Fischer 344 x Brown Norway rats as nine fractions over 4.5 weeks leads to impaired Morris water maze (MWM) performance 12 months later. Compared to sham-irradiated rats, the irradiated rats demonstrated impaired MWM performance. The relative levels of the NR1 and NR2A but not the NR2B subunits of the NMDA receptor were significantly higher in hippocampal CA1 of irradiated rats compared to control rats. No significant differences were detected for these NMDA subunits in CA3 or dentate gyrus. Further analysis of CA1 revealed that the relative levels of the GluR1 and GluR2 subunits of the AMPA receptor and synaptophysin were not altered by whole-brain irradiation. In summary, a clinically relevant regimen of fractionated whole-brain irradiation led to significant impairments in spatial learning and reference memory and alterations in the relative levels of subunits of the NMDA, but not the AMPA, receptors in hippocampal CA1. These findings suggest for the first time that radiation-induced cognitive impairments may be associated with alterations in glutamate receptor composition.     

A comparison of paternal behaviour in the meadow vole Microtus pennsylvanicus,the pine vole M. pinetorum and the prairie vole M. cchrogaster

Paternal care in microtines has been studied infrequently and few studies have compared patterns of direct and indirect paternal investment. The paternal behaviour of three vole species, the meadow vole (Microtus pennsylvanicus), the pine vole (M. pinetorum) and the prairie vole (M. ochrogaster) was examined in a semi-natural setting. Prairie and pine voles were found to exhibit high levels of paternal care. Prairie vole males contributed the most direct care by remaining in the natal nest for long periods of time in contact with the pups. Pine voles contributed less direct care than prairie voles as they spent less time in the natal nest with their offspring. In addition, both prairie and pine vole males were observed to groom their pups and retrieve them back to the nest area. Prairie vole males also engaged in such indirect forms of care as nest construction and maintenance, while pine voles provided indirect care in the form of tunnel construction and food caching. Meadow vole males were the least paternal of the three species and rarely engaged in either direct or indirect care. These findings support predictions that M. pennsylvanicus is promiscuous and that male and female meadow voles occupy separate territories. They are also consistent with studies which indicate that prairie and pine voles are monogamous and have a structured social organization with members interacting closely with one another.     

Evidence for the involvement of d-aspartic acid in learning and memory of rat

d-Aspartic acid (d-Asp) is an endogenous amino acid present in neuroendocrine systems. Here, we report evidence that d-Asp in the rat is involved in learning and memory processes. Oral administration of sodium d-aspartate (40 mM) for 12–16 days improved the rats’ cognitive capability to find a hidden platform in the Morris water maze system. Two sessions per day for three consecutive days were performed in two groups of 12 rats. One group was treated with Na-d-aspartate and the other with control. A significant increase in the cognitive effect was observed in the treated group compared to controls (two-way ANOVA with repeated measurements: F _{(2, 105)} = 57.29; P value < 0.001). Five further sessions of repeated training, involving a change in platform location, also displayed a significant treatment effect [F _{(2, 84)} = 27.62; P value < 0.001]. In the hippocampus of treated rats, d-Asp increased by about 2.7-fold compared to controls (82.5 ± 10.0 vs. the 30.6 ± 5.4 ng/g tissue; P < 0.0001). Moreover, 20 randomly selected rats possessing relatively high endogenous concentrations of d-Asp in the hippocampus were much faster in reaching the hidden platform, an event suggesting that their enhanced cognitive capability was functionally related to the high levels of d-Asp. The correlation coefficient calculated in the 20 rats was R = −0.916 with a df of 18; P < 0.001. In conclusion, this study provides corroborating evidence that d-aspartic acid plays an important role in the modulation of learning and memory.     

Spatial memory in the rat requires the dorsolateral band of the entorhinal cortex

The extensive connections of the entorhinal cortex with the hippocampus and the neocortex point to this region as a major interface in the hippocampal-neocortical interactions underlying memory. We asked whether hippocampal-dependent recall of spatial memory depends on the entorhinal cortex, and, if so, which parts are critical. After training in a Morris water maze, rats received fiber-sparing lesions in the dorsolateral band of the entorhinal cortex, which mediates much of the visuospatial input to the dorsal hippocampus. These lesions entirely disrupted retention and retarded new learning. Spatial memory was spared by lesions in the ventromedial band, which connects primarily with ventral hippocampus, but these lesions reduced defensive behavior on an elevated plus maze, mirroring the effects of damage to ventral hippocampus. The results suggest that the functional differences between dorsal and ventral hippocampus reflect their connectivity with modules of the entorhinal cortex that are differently linked to the rest of the cortex.     

Spontaneous bacterial cell lysis and biofilm formation in the colon of the Cape Dune mole-rat and the laboratory rabbit

A wide range of techniques, including high-throughput DNA sequencing methods, have been applied to the evaluation of the normal intestinal flora. However, the inability to grow many of those species in culture imposes substantial constraints on the techniques used to evaluate this important community. The presence of biofilms in the normal gut adds further complexity to the issue. In this study, a flow cytometric analysis was used to separate intact bacterial cells, cell debris, and other particulate matter based on bacteria-specific staining and particle size. In addition, an analysis of biofilm formation using fluorescent light microscopy was conducted. Using these approaches, the ratio of bacterial cell debris to intact bacterial cells as a measure of spontaneous lysis of bacterial cells in the gut of the Cape dune mole-rat (Bathyergus suillus) and the laboratory rabbit (Oryctolagus cuniculus) was examined, and the degree of biofilm formation was semi-quantitatively assessed. The results suggest that the degree of spontaneous cell lysis was greater in the appendix than in the cecum in both the mole-rat and the rabbit. Further, the results point toward extensive epithelial-associated biofilm formation in the proximal mole-rat and rabbit large bowel, although the biofilms may be less structured than those found in laboratory rodents and in humans.     

Spinal cord development in marsupials in relation to birthing strategies and in comparison with monotremes and the laboratory rat

Marsupials are born in an immature state, followed by prolonged nurturing of pouch young by maternal lactation. Spinal cord sections held in the collections at the Museum für Naturkunde, Berlin were used to test the relationship between structural maturity of the spinal cord and the locomotor challenges that face young marsupials and monotremes. Analysis of variance indicated that body length is a much stronger determinant of variation in anatomical measures of spinal cord maturation than mammal type.     

Spatial learning induces predominant downregulation of cytosolic proteins in the rat hippocampus

Spatial learning is known to depend on protein synthesis in the hippocampus. Whereas the role of the hippocampus in spatial memory is established, the biochemical and molecular mechanisms underlying this process are poorly understood. To comprehend the complex pattern of protein expression induced by spatial learning, we analyzed alterations in the rat hippocampus proteome after 7 days of spatial learning in the Morris water maze. Forty Wistar rats were randomized into two groups. Animals of group A learned to localize a hidden platform in the water maze. Animals of group B served as controls and spent exactly the same time in the water maze as animals of group A. However, no platform was used in this test and the rats could not learn to localize the target. After the last trial, hydrophilic proteins from the hippocampus were isolated. A proteome-wide study was performed, based on two-dimensional gel electrophoresis and mass spectrometry. Compared with non-learning animals, 53 (70%) proteins were downregulated and 23 (30%) proteins were upregulated after 7 days in rats with spatial learning. The overall changes in protein expression, as quantified by the induction factor, ranged from -1.62 (downregulation to 62%) to 2.10 (upregulation by 110%) compared with controls (100%). Most identified proteins exhibit known functions in vesicle transport, cytoskeletal architecture, and metabolism as well as neurogenesis. These findings indicate that learning in the Morris water maze has a morphological correlate on the proteome level in the hippocampus.     

Hypoxia associated NMDA receptor 2 subunit composition: developmental comparison between the hypoxia-tolerant subterranean mole-rat, Spalax, and the hypoxia-sensitive rat

Vertebrate brains are sensitive to oxygen depletion, which may lead to cell death. Hypoxia sensitivity originates from the high intrinsic rate of ATP consumption of brain tissue, accompanied by the release of glutamate, leading to the opening of ionotropic glutamate receptors, such as N-methyl-D-aspartate (NMDA) receptors (NMDARs). The relative expression levels of the four NMDAR-2 (NR2) subunits change during mammalian development with higher levels of units NR2B and NR2D observed during early development and correlated with hypoxic tolerance during embryonic and neonatal stages of development. Higher levels of NR2D are also abundant in brains of hypoxia tolerant species such as the crucian carp. The subterranean mole-rat, Spalax spends its life underground in sealed burrows and has developed a wide range of adaptations to this special niche including hypoxia-tolerance. In this study, we compared the in vivo mRNA expression of NR2 subunits in the brains of embryonic, neonatal and adult Spalax and rat. Our results demonstrate that under normoxic conditions, mRNA levels of NR2D are higher in Spalax than in rat at all developmental stages studied and are similar to levels in neonatal rat and in other hypoxia/anoxia tolerant species. Furthermore, under hypoxia Spalax NR2D mRNA levels increase while no response was observed in rat. Similarly, hypoxia induces an increase in mRNA levels of Spalax NR2A, claimed to promote neuronal survival. We suggest that indeed the proportional combinations of NMDAR-2 subunits contribute to the ability of the Spalax brain to cope with hypoxic environments.