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Clearly the brain controls behavior but can behavior also “control” the brain? On an evolutionary time scale, selective ecological pressures shape the sensory and motor capacities as well as the body and behavior. Correspondingly, in development, behavior acts in concert with the environment to cause structural changes in the brain lasting a lifetime. Surprisingly, in “real time” social behavior can also cause changes, typically reversible, in the brain in adult animals. Changes caused by behavioral interactions can be dramatic, and in many instances, these interactions are directly related to reproductive behavior. Understanding how behavior sculpts the brain in the course of behavioral interactions is a major challenge. Analyzing such changes requires a model system allowing control of the biological and behavioral environment of many animals simultaneously yet allowing access to physiological, cellular and molecular processes being regulated. The mouthbrooding cichlid Haplochromis (Astatotilapia) burtoni (Günther) from Lake Tanganyika lends itself to the study of social influences on the brain. It has complex, though easily observable individual and social behaviors regulated by two distinct classes of males, those with territories and those without. Many features of the animals are shaped by social encounters including the maturation of juveniles, the hypothalamic-pituitary-gonadal axis, the growth rate, the basal stress level among others. How does social information effect change in the brain and body? Animals must attend to the social scene to identify their chances. Learning how social information is transduced into cellular changes in this species should help understand how this happens in other social animals.
Microarray technology is a powerful technique that allows the simultaneous study of thousands of gene transcripts. During the past two years there has been an explosion of publications describing experiments utilizing microarray technology that range from original research findings from biological paradigms to mathematically modeled systems. However, neuroscientists using microarray technology face significant challenges due to high tissue complexity, low abundance transcripts, and small magnitude changes in transcript levels that have significant biological impact. This manuscript describes a series of studies designed to address issues regarding microarray sensitivity, ability of microarrays to detect subtle changes, and reproducibility of microarray experiments, all in the context of neuronal tissue. From the presentation of these studies, the authors argue that although microarray technology is limited with regards to sensitivity, the outcome of these experiments, if approached with appropriate skepticism, can be fruitful in the generation of hypotheses and seeding of future experiments.
Over the last decade, subtractive cloning approaches have been used extensively to isolate genes that are up- or down-regulated under various conditions. These techniques have provided the foundation for many subsequent studies concerning gene function and regulation and, as such, have been valuable tools for many biological fields. Over the past 10 years, we have used different subtractive cloning approaches to isolate genes in fish that are regulated in relation to hormonal stimulation or the stage of ovarian maturation. These include conventional cDNA subtraction followed by library screening, differential display PCR, suppression subtraction hybridization, and more recently, iterative PCR subtraction. We continue to use these techniques for the isolation of new genes involved in physiological processes in fish and bivalve molluscs. Examples that illustrate the use of these different subtractive cloning techniques are described, including where possible the advantages and disadvantages of each. In addition, the use of ancillary methods (e.g., “Reverse Northerns”) to facilitate the use of these subtractive approaches are discussed.
Many studies suggest that migratory guidance cues within the developing brain are diverse across many regions. To better understand the early development and differentiation of select brain regions, an in vitro method was developed using selected inbred and transgenic strains of embryonic mice. In particular, organotypic slices are used to test factors that influence the movements of neurons during brain development. Thick 250 μm slices cut on a vibrating microtome are prepared and maintained in vitro for 0–3 days. Nissl stain analyses often show a uniform distribution of cells in the regions of interest on the day of plating (embryonic days 12–15). After 3 days in vitro, cellular aggregation suggesting nuclear formation or the changing position of cells with a defined phenotype show that reasonably normal cell movements occur in several regions. Movements in vitro that mimic changes in vivo suggest that key factors reside locally within the plane of the slices. Video microscopy studies are used to follow the migration of fluorescently labeled cells in brain slices from mice maintained in serum-free media for 1 to 3 days. Transgenic mice with selective promoter driven expression of fluorescent proteins allow us to view specific cell types (e.g., neurons expressing gonadotropin-releasing hormone). The accessibility of an in vitro system that provides for relatively normal brain development over key brief windows of time allows for the testing of important mechanisms.