LTP: variation between inbred mouse strains.
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Animals↗
Biomedical subjects
R Gerlai
Publications and source records attributed to R Gerlai.
Alzheimer's disease: beta-amyloid hypothesis strengthened!
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Alzheimer Disease↗
Eph receptors and neural plasticity.
Eph receptor tyrosine kinases are largely known for their involvement in brain development but, as some of these receptor tyrosine kinases are also expressed in adults, their possible role in the mature nervous system has begun to be explored. Evidence for the involvement of Eph receptors in synaptic plasticity, learning and memory is only emerging and needs corroboration. However, it is likely that the actions of Eph kinases in the adult brain will attract significant attention and become a fertile research area, as occurred in the case of the neurotrophins.
Animals↗
Drinks like a fish: zebra fish (Danio rerio) as a behavior genetic model to study alcohol effects.
Zebra fish may be an ideal vertebrate model system for numerous human diseases with which the genetics and biological mechanisms of the disease may be studied. Zebra fish has been successfully used in developmental genetics, and recently, neurobiologists have also started to study this species. A potentially interesting target disease amenable for analysis with zebra fish is drug addiction, e.g. alcoholism. Although genetic tools to manipulate the genome of zebra fish are available, appropriate phenotypical testing methods are often lacking. In this paper, we describe basic behavioral tests to investigate the acute effects of alcohol on zebra fish. These behavioral paradigms will be useful for the genetic and biological analysis of acute and chronic drug effects as well as addiction. In addition to presenting findings for the acute effects of alcohol, we briefly describe our strategy for generating and screening mutants. We hope that our pilot work will facilitate the future development of behavioral tests and the use of zebra fish in the genetic analysis of the biological effects of drugs of abuse.
Aggression↗
Protein targeting: altering receptor kinase function in the brain.
Gene targeting has proved to be one of the most powerful techniques with which one can investigate molecular mechanisms that underlie complex phenomena such as learning and memory. Despite its popularity, however, concerns have been raised about this technique and alternative approaches have been sought. One such approach is protein targeting, which is based on the application of immunoadhesins, genetically engineered fusion proteins that exhibit functionally relevant target specificity. These immunoadhesins modulate the activity of not only a single receptor but of all receptors with homologous binding sites, which thereby eliminates the possibility of compensation by sister receptors. Furthermore, immunoadhesins can be used not only to impair but also to improve receptor function in the brain. Initial studies using immunoadhesins suggest that protein targeting might be a useful approach for analyzing the molecular mechanisms of brain function and behavior.
Animals↗
Transient focal cerebral ischemia induces sensorimotor deficits in mice.
Rodents have been extensively used for experimental stroke research with rat and gerbil the preferred species. With the advent of transgenesis and gene targeting the number of mutant mouse strains is rapidly increasing. Thus, mouse models of stroke will be of great importance in the analysis of genetic factors affecting stroke. Demonstrating long-term functional recovery is of paramount importance for the pharmacological evaluation of putative stroke therapies. In the present paper we induce mild focal cerebral ischemia by tandem occlusion of the right middle cerebral artery (MCA), via craniotomy, together with the common carotid artery for 45 min in C57BL/6 strain of mice. The effects of ischemia were evaluated acutely by MRI and long-term (> 3 weeks) sensorimotor functional deficits were analyzed using a number of behavioral paradigms including the rotorod, wire hang, horizontal surface approach, eye-closure reflex, and T-maze tests. Although the induced brain damage is mild we show that it leads to clearly detectable and significant sensorimotor defects associated with fine motor coordination, balance, and postural and sensory reflexes. We conclude that the applied behavioral tests will be useful in the analysis of stroke in mutant mice.
Animals↗
Anesthesia induced retrograde amnesia is ameliorated by ephrinA5-IgG in mice: EphA receptor tyrosine kinases are involved in mammalian memory.
EphA receptors and their ephrin-A ligands were previously thought to play a role only in embryonic development of the brain. Recently, however, these proteins were shown to be expressed in the adult mouse brain, primarily in the hippocampus, and were implicated in hippocampal synaptic plasticity and learning. What aspects of learning EphA receptors mediate have not been studied? Using the fear conditioning paradigm we demonstrate that EphA receptors play roles in memory. We show that post-training surgical anesthesia leads to robust context specific retrograde amnesia in mice, and post-anesthesia activation of EphA receptors induces a significant amelioration of this amnesia. As acquisition was left unaffected and performance factors were found unaltered, we suggest that the amelioration was due to changes in cognition leading to improved memory. Our data represent the first pieces of evidence for the involvement of EphA receptor tyrosine kinase receptors in mammalian memory, a finding that opens a new avenue into the functional analysis of the largest receptor tyrosine kinase subfamily in the brain.
Amnesia, Retrograde↗
Heregulin, but not ErbB2 or ErbB3, heterozygous mutant mice exhibit hyperactivity in multiple behavioral tasks.
Genetic redundancy is a problem in gene targeting studies because functionally relevant sister proteins can compensate for the lack of protein product of a targeted gene. A molecular system is chosen in which it is hoped to demonstrate both the lack and presence of compensation after disruption of particular single genes. Mammals may not be able to compensate for the lack of heregulin, a single ligand for multiple ErbB receptors, however, compensation is expected when a single ErbB receptor is knocked out. To investigate this the heregulin-1, ErbB2, or ErbB3 locus was disrupted in a targeted manner and mice heterozygous for the mutation were analyzed. Heregulin and its receptors were shown to be involved in embryonic brain development and, more recently, in plastic changes associated with adult brain function in rodents. Although they have never been shown to play roles in mammalian behavior, it was decided to characterize the mice behaviorally using a battery of simple tests. Heregulin mutant mice exhibited elevated activity levels in the open field, showed improved rotorod performance, and finished T-maze spontaneous alternation task faster compared to control wild type littermates, findings that suggest a consistent hyperactivity across tests. ErbB2 and ErbB3 mutant mice, whose strain origin was identical to that of heregulin mutants, showed no sign of the behavioral alterations. It is suggested that the abnormalities seen in heregulin mutant mice are due to mutation at that locus and the lack of alterations seen in ErbB2 and ErbB3 mutant mice is the result of compensation by unaltered sister receptors.
Animals↗
Targeting genes and proteins in the analysis of learning and memory: caveats and future directions.
Gene targeting using homologous recombination in embryonic stem (ES) cells and transgenic approaches in general allow one to precisely manipulate single genes and investigate their in vivo function in the mouse. Geneticists argue that these techniques are superior to pharmacological approaches as they obviate the lack of highly specific pharmacological agents in the study of brain function and behavior. However, by now it has become clear that transgenic approaches also have some limitations. One problem is spatial and temporal specificity of the genetic manipulation. The other is the possibility that the introduced genetic alteration gives rise to complex, secondary phenotypic changes. This may be a disadvantage in the functional analysis of genes associated with learning and memory especially if the gene of interest plays roles in embryonic development of the brain as well as in adult neural function. Examples of such genes include, but are not limited to, those encoding neurotrophins, cell adhesion molecules, and protein kinases. Second generation gene targeting with inducible and cell type restricted knock-out, or transgenic approaches with inducible gene expression systems, will solve some problems. However, at present these strategies also suffer from difficulties inherent to the traditional knock-out. Several strategies alternative to transgenic approaches are also available. Antisense oligonucleotides, antibodies, or pharmacological agents may be used to manipulate molecular events at the transcription, translation, or protein function levels. I review these strategies briefly and suggest yet another approach: protein targeting with the use of recombinant immunoadhesins. I suggest that this latter approach has the specificity of gene targeting but lacks some of its disadvantages.
Animals↗
Regulation of learning by EphA receptors: a protein targeting study.
EphA family receptor tyrosine kinases and their ephrin-A ligands are involved in patterning axonal connections during brain development, but until now a role for these molecules in the mature brain had not been elucidated. Here, we show that both the EphA5 receptor and its ephrin-A ligands (2 and 5) are expressed in the adult mouse hippocampus, and the EphA5 protein is present in a phosphorylated form. Because there are no pharmacological agents available for EphA receptors, we designed recombinant immunoadhesins that specifically bind to the receptor binding site of the ephrin-A ligand (antagonist) or the ligand binding site of the EphA receptor (agonist) and thus target EphA function. We demonstrate that intrahippocampal infusion of an EphA antagonist immunoadhesin leads to impaired performance in two behavioral paradigms, T-maze spontaneous alternation and context-dependent fear conditioning, sensitive to hippocampal function, whereas activation of EphA by infusion of an agonist immunoadhesin results in enhanced performance on these tasks. Because the two behavioral tasks have different motivational, perceptual, and motor requirements, we infer the changes were not caused by these performance factors but rather to cognitive alterations. We also find bidirectional changes in gene expression and in electrophysiological measures of synaptic efficacy that correlate with the behavioral results. Thus, EphA receptors and their ligands are implicated as mediators of plasticity in the adult mammalian brain.
Animals↗
Evidence for a protective role of metallothionein-1 in focal cerebral ischemia.
Metallothioneins (MTs) are a family of metal binding proteins that have been proposed to participate in a cellular defense against zinc toxicity and free radicals. In the present study, we investigated whether increased expression of MT in MT-1 isoform-overexpressing transgenic mice (MT-TG) affords protection against mild focal cerebral ischemia and reperfusion. Transient focal ischemia was induced in control (wild type) and MT-TG mice by occluding the right middle cerebral artery for 45 min. Upon reperfusion, cerebral edema slowly developed and peaked at 24 hr as shown by T2-weighted MRI. The volume of affected tissue was on the average 42% smaller in MT-TG mice compared with control mice at 6, 9, 24, and 72 hr and 14 days postreperfusion (P < 0.01). In addition, functional studies showed that 3 weeks after reperfusion MT-TG mice showed a significantly better motor performance compared with control mice (P = 0.011). Although cortical baseline levels of MT-1 mRNA were similar in control and MT-TG mice, there was an increase in MT-1 mRNA levels in the ischemic cortex of MT-TG mice to 7.5 times baseline levels compared with an increase to 2.3 times baseline levels in control mice 24 hr after reperfusion. In addition, MT-TG mice showed an increased MT immunoreactivity in astrocytes, vascular endothelial cells, and neurons 24 hr after reperfusion whereas in control mice MT immunoreactivity was restricted mainly to astrocytes and decreased in the infarcted tissue. These results provide evidence that increased expression of MT-1 protects against focal cerebral ischemia and reperfusion.
Animals↗
Analysing hippocampal function in transgenic mice: an ethological perspective.
Advances in molecular genetics and technology have led to the dawn of a new era for neuroscience: manipulation of single genes now makes it possible to dissect the complexities of neurobiological phenotypes and to understand many of the intricacies of brain and behaviour, even in mammals. The phenotypical analysis of these mutant animals is complicated because the potential outcome of gene manipulation is difficult to predict. While behavioural analysis should form an integral part of any multidisciplinary research programme investigating the phenotypical effects of single genes on hippocampal function, it is crucial that the behavioural tests are designed and conducted appropriately. Approaches that take species-specific behavioural characteristics into account and use ethological methods could be the most useful for interpreting these behavioural findings and understanding the biological mechanisms of brain function.
Animals↗
VEGF antagonism reduces edema formation and tissue damage after ischemia/reperfusion injury in the mouse brain.
VEGF is mitogenic, angiogenic, and a potent mediator of vascular permeability. VEGF causes extravasation of plasma protein in skin bioassays and increases hydraulic conductivity in isolated perfused microvessels. Reduced tissue oxygen tension triggers VEGF expression, and increased protein and mRNA levels for VEGF and its receptors (Flt-1, Flk-1/KDR) occur in the ischemic rat brain. Brain edema, provoked in part by enhanced cerebrovascular permeability, is a major complication in central nervous system pathologies, including head trauma and stroke. The role of VEGF in this pathology has remained elusive because of the lack of a suitable experimental antagonist. We used a novel fusion protein, mFlt(1-3)-IgG, which sequesters murine VEGF, to treat mice exposed to transient cortical ischemia followed by reperfusion. Using high-resolution magnetic resonance imaging, we found a significant reduction in volume of the edematous tissue 1 day after onset of ischemia in mice that received mFlt(1-3)-IgG. 8-12 weeks after treatment, measurements of the resultant infarct size revealed a significant sparing of cortical tissue. Regional cerebral blood flow was unaffected by the administration of mFlt(1-3)-IgG. These results demonstrate that antagonism of VEGF reduces ischemia/reperfusion-related brain edema and injury, implicating VEGF in the pathogenesis of stroke and related disorders.
Animals↗
Protein targeting in the analysis of learning and memory: a potential alternative to gene targeting.
Gene targeting using homologous recombination in embryonic stem (ES) cells offers unprecedented precision with which one may manipulate single genes and investigate the in vivo effects of defined mutations in the mouse. Geneticists argue that this technique abrogates the lack of highly specific pharmacological tools in the study of brain function and behavior. However, by now it has become clear that gene targeting has some limitations too. One problem is spatial and temporal specificity of the generated mutation, which may appear in multiple brain regions or even in other organs and may also be present throughout development, giving rise to complex, secondary phenotypical alterations. This may be a disadvantage in the functional analysis of a number of genes associated with learning and memory processes. For example, several proteins, including neurotrophins--cell-adhesion molecules--and protein kinases, that play a significant developmental role have recently been suggested to be also involved in neural and behavioral plasticity. Knocking out genes of such proteins may lead to developmental alterations or even embryonic lethality in the mouse, making it difficult to study their function in neural plasticity, learning, and memory. Therefore, alternative strategies to gene targeting may be needed. Here, we suggest a potentially useful in vivo strategy based on systemic application of immunoadhesins, genetically engineered fusion proteins possessing the Fc portion of the human IgG molecule and, for example, a binding domain of a receptor of interest. These proteins are stable in vivo and exhibit high binding specificity and affinity for the endogenous ligand of the receptor, but lack the ability to signal. Thus, if delivered to the brain, immunoadhesins may specifically block signalling of the receptor of interest. Using osmotic minipumps, the protein can be infused in a localized region of the brain for a specified period of time (days or weeks). Thus, the location and timing of delivery are controlled. Here, we present methodological details of this novel approach and argue that infusion of immunoadhesins will be useful for studying the role particular receptors play in behavioral and neural plasticity.
Animals↗
Contextual learning and cue association in fear conditioning in mice: a strain comparison and a lesion study.
Fear conditioning with electric shock (unconditioned stimulus, US) paired with tone cue (conditioned stimulus, CS) has been extensively applied in recent molecular neurobiological analysis of hippocampal dysfunction in mice because the context-dependent test phase of this learning paradigm is claimed to detect hippocampal impairment in a specific manner, whereas the cue-dependent test serves as a control situation independent of hippocampal function. These claims are based on hippocampal lesion studies performed with rats and have not been conclusively confirmed with mice with specific hippocampal lesion. Therefore, I investigated how hippocampal ibotenic acid lesion affects conditioned fear in mice. I confirm that extensive lesions localized to the hippocampus impair context-dependent learning but also show that, unlike in the original rat studies, the behavioral impairment is only partial. Furthermore, studying two inbred strains of mice (C57BL/6 and DBA/2) with highly different hippocampal function, I show that the presence or absence of CS during training may influence the mouse's ability to learn complex multiple contextual stimuli in a genotype-dependent manner. I conclude that performance at the 'context' test may be based on complex configural (hippocampal) learning but it can also be based on a more simple elemental (non-hippocampal) learning thus leading to potentially false-negative findings in the analysis of hippocampal dysfunction.
Acoustic Stimulation↗
A new continuous alternation task in T-maze detects hippocampal dysfunction in mice. A strain comparison and lesion study.
The mammalian hippocampus has been the focus of several neurobiology studies because of its important behavioral function and because long-term potentiation (LTP) is a prominent feature of this brain region. Converging evidence suggests that hippocampal function is associated with learning multiple relationships of environmental cues. In this paper a novel behavioral test procedure is introduced, a modified T-maze continuous alternation task (T-CAT), that may serve as a simple, automatable, and quick test of hippocampal function in addition to the frequently applied water maze and fear conditioning paradigms. A comparison is made between mice (strain C57BL/6) with ibotenic acid lesioned or vehicle injected hippocampus, two transgenic strains (on CD1 background) overexpressing a calcium binding protein, S100beta, and inbred (C57BL/6, DBA/2, 129/SV and 129/SVEV) and outbred (CD1) strains of mice. This study shows that hippocampal lesioning led to a significant impairment in T-CAT. Furthermore, overexpression of S100beta, which impairs hippocampal LTP, also led to an impairment demonstrating that T-CAT is sensitive to detect hippocampal dysfunction. Analysis of the mouse strains revealed that C57BL/6 and CD1 mice performed well in T-CAT, whereas 129/SV, 129/SVEV and DBA/2 were significantly impaired, a finding that underscores the importance of strain differences in pharmacological or single gene manipulation studies of hippocampal function in mice.
Animals↗