Implementing iDISCO+ for mapping c-fos expression in insular cortex after object recognition training and identifying neuronal networks after rAVV2-retro injection

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2017-08-01
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en
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Abstract
The role of the insular cortex in emotional memory has been recently acquired significance. Its interaction with BLA regarding to emotionally enhanced memories gained the spotlight after its consistent effect shown in human brain imaging studies. However, the involvement of the IC depending on its complex anatomical architecture and its mediation on the function of emotional memory remain largely unknown. In this thesis, I investigated the differential c-fos expression in the IC after different training groups which performed object recognition task. Possible changes in the c-fos expression within groups and distinct sub-layers of the insular cortex. In the procedure, I specifically focused on anterior and posterior part and differential expression. In addition to looking at granular, dysgranular and agranular insular cortex. For the investigation of the behavioral groups, I have used a novel brain clearing technique called iDISCO+ which enables me to pinpoint the anatomical structures in 3D environment and look at the differential expression exclusively. In the retrograde virus injected brain, it has also been processed by using iDISCO+ because it preserves endogenous florescence proteins such as GFP and mCherry. It exhibits good signal to noise ratio and deep tissue penetration in an acceptable time period. It was suitable method to see the c-fos expression in anterior and posterior insular cortex and as well as amygdala and PFC. Also, used to observe mCherry expression after rAAV2-retro injection to see retrograde labeling of the neuron projection. The experimental setup aimed to unravel the structural connections of the IC network by using the object recognition training as its c-fos inducer and tracing the neuronal activity by retrograde tracer virus. All, these points are explored by the application of a novel brain clearing technique.
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Faculteit der Sociale Wetenschappen