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    • Fluorescein TSA Fluorescence System Kit br Methods br Acknow

    2021-12-15


    Methods
    Acknowledgements The authors thank the ZLS group members, particularly Li Li and Elijah Roberts, for many helpful discussions. They also wish to thank Nathan Baker for APBS assistance, Jan Jensen for help with PROPKA 2.0, Susan Martinis for experimental interpretations, and John Stone for VMD graphics suggestions. Funding for A.B.P., J.E., and A.S. was provided by National Science Foundation grants MCB04-46227, MCB08-44670, and PHY08-22613, and by National Institutes of Health Chemical Biology Training Grant (5T32GM070421). Supercomputer and local computing time were provided by National Center for Supercomputing Applications Large Resource Allocations Committee grant MCA03T027 and National Science Foundation Chemistry Research Instrumentation and Facilities grant 0541659.
    Introduction Over the years, studies of learning and memory processes in animals have employed a great many experimental approaches and techniques, and have used a range of different species. In the past 15 years the rapid advances in genetic engineering and molecular biological techniques have provided a unique and extremely powerful tool for analyzing these processes in rodents and, in particular, in mice (Chen and Tonegawa, 1997). The study of genetically modified mice has revealed dissociations in aspects of information processing that were hitherto unapparent on the basis of more traditional experimental approaches such as lesion studies. The use of these transgenic approaches has been particularly successful for studying the contribution that different glutamate receptors and their specific subunits make to distinct aspects of learning and memory. Glutamatergic neurotransmission underlies the majority of fast, synaptic neurotransmission in the Fluorescein TSA Fluorescence System Kit and also plays a crucial role in plasticity processes by which the efficacy of synaptic connections between neurons can be strengthened or weakened. These plasticity mechanisms are widely considered to play an essential role in learning and memory (Hebb, 1949; Bliss and Lomo, 1973; Morris et al., 1986a; Martin et al., 2000). Glutamatergic neurotransmission in the CNS is mediated through both metabotropic and ionotropic receptors. There are three main forms of ionotropic glutamate receptor —l-α-amino-3-hydroxy-5-methyl-4-isoxazelopropionate (AMPA), N-methyl-d-aspartate (NMDA) and kainate — characterised by their distinct anatomical localizations within the brain, differential sensitivity to a variety of pharmacological antagonists and their selective activation by various glutamate analogues, which provide the basis for their nomenclature (Wisden and Seeburg, 1993; Sprengel, 2006). Although the role of the NMDA receptor in learning and memory has been exhaustively studied and extensively described (Morris et al., 1990a; Martin et al., 2000; Martin and Morris, 2002), the contribution that AMPA receptors, and their individual subunits, make to these aspects of information processing has been less well studied, in part as a consequence of the lack of the appropriate pharmacological tools. Recent advances in genetic technologies now allow the importance of the individual receptor subunits to be assessed. The functional significance of a particular receptor subunit or subtype will obviously depend upon the brain region and extended neural circuitry within which it is embedded. This chapter aims to describe and discuss the role of AMPA receptors, and in particular GluR-A (GluR1)-containing AMPA receptors, in hippocampus-dependent forms of learning and memory. GluR-A-containing AMPA receptors in other regions of the brain are likely to play an equally important role in other aspects of behaviour. For example, the role of the GluR-A subunit in encoding and/or retrieving the sensory-specific aspects of unconditioned stimuli has now been established (Mead and Stephens, 2003; Johnson et al., 2005; Rumpel et al., 2005), a phenotype that bears strong resemblance to the effects of cytotoxic, basolateral amygdala lesions (Blundell et al., 2001; Dwyer and Killcross, 2006). This chapter will, however, focus on the hippocampal memory system.