Download PDF Molecular Biology of the Neuron (Molecular and Cellular Neurobiology) Second Edition by R. W. Davies

Sinopsis

The detailed study of neuronal function, in particular the function of neurons involved in complex processing and behavioural roles, requires several factors to be controlled, measured or exploited. Of relevance to the study of neuronal function are:

1. Drosophila: the short life cycle and simple culture conditions make it easy to control external environmental variables which are often more problematic in other species. In particular, the ‘sensory’ and ‘social’ interactions of the organism can easily be monitored and/or manipulated directly.
2. Behaviour: in order to unravel the neuronal pathways underlying behaviour, it is essential to study behaviours that can be assayed both qualitatively and quantitatively. There is a range of assays that have been developed over the years for both larval and adult stages of Drosophila. These include the use of visual, olfactory, tactile and auditory cues and assessment of responses with respect to reflex reactions as well as more complex learned behaviour.
3. Neurogenetics: Drosophila is one of the longest established genetic models with a formidable array of genetic and molecular tools that can be brought to bear upon any biological research area. This organism has given us valuable insights into the molecular, cellular and evolutionary bases of behaviour. In this chapter we will restrict our attention to those particularly relevant to neuroscience research in the organism.
4. Nervous system: the detailed neuronal architecture of the nervous system needs to be known in order for pathways involving multiple neurons to be investigated; i.e. neurons act as parts in circuits and context is critical. For Drosophila, traditional neuroanatomical techniques have produced gross maps of the nervous system through development and have also contributed valuable information about single neuronal projection patterns. The use of gene expression profiles (via enhancertraps) to unravel neuroanatomy was pioneered in Drosophila and recent techniques have refined such approaches to the extent that brain maps for Drosophila will soon be available to single neuron resolution.
5. Neurophysiology: sadly the small size of neurons in Drosophila makes traditional neurophysiology difficult. However, there have been advances recently with several emerging techniques that allow non-invasive recording of neuronal activity.
6. Neuroinformatics and high-throughput technologies, supporting informatics analysis toolkits and public database systems. Historically, Drosophila has had one of the most up-to-date and comprehensive databases describing the genetics of the organism (FlyBase: http://flybase.bio. indiana.edu) and this has been supplemented by a range of genome databases describing the sequences generated by the public and private genome sequencing projects and their annotation. In addition there are additional databases and resources of specific interest to Drosophila neuroscience that will be described towards the end of this chapter.

Drosophila melanogaster

Drosophila melanogaster (referred to here as Drosophila) is generally the first multicellular experimental organism that biologists are introduced to in their training. Unlike most other experimental systems in biology, their simple culture requirements are so simple and cost-effective that they can be maintained in high-school biology labs with ease. In a modern laboratory situation this makes getting started with Drosophila particularly easy, although as for any organism large-scale operations will require some dedicated equipment and support staff.
 

For the newcomer, there is a wealth of literature describing the culture requirements and the basic (and advanced) techniques possible. To begin with, we would recommend ‘Drosophila protocols’ (Sullivan et al. 2000), which is a laboratory manual (in format and weight). It describes in detail many of the common laboratory techniques including many of the recent molecular/transgenic manipulations used in Drosophila neuroscience, some of which we will describe in more detail here.

Content

1. Studying neuronal function using the Drosophila genetic system
2. Using mouse genetics to study neuronal development and function
3. Gene expression: from precursor to mature neuron
4. Protein trafficking in neurons
5. Ion channels and electrical activity
6. Molecular biology of transmitter release
7. Molecular biology of postsynaptic structures
8. Signal reception: Ligand-gated ion channel receptors
9. Signal reception: G protein-coupled receptors
10. Synapse-to-nucleus calcium signalling
11. Signalling by tyrosine phosphorylation in the nervous system
12. Mature neurons: Signal transduction-serine/ threonine kinases
13. The cytoskeleton
14. Neuronal Plasticity
15. Genetic basis of human neuronal diseases
16. Ageing and the death of neurones

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