Hippocampus Case Study

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5.1 Introduction

In the brain, the hippocampus is involved in directing the process of creating, systematising and retaining memories. The hippocampus is widely connected to the dorso medial nuclei of the thalamus, mammillary nuclei of the hypothalamus, limbic system network for learning and continuous action for long-term storage. The hippocampus brings about the representation of spatial and temporal memories (Eichenbaum et al, 1992). The research about cognitive function of the hippocampus suggests that selective lesions of the hippocampus will produce a deficit in learning or exploration of a new environment. In the hippocampus memory, related information passes through various regions like cornu ammonis (CA) and dentate gyrus (DG).
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The brain tissue was harvested and placed in buffered formalin solution. After fixation, the tissue sample was washed thoroughly in running water overnight. It was dehydrated by ascending rates of alcohol (50, 60, 70, 80, 90 and 100%) each for 60 minutes. Then the tissues were cleared with two changes of xylene each with 30 minutes. Embedding was done with two changes of molten paraffin for 30 min to remove xylene. They were mounted in L blocks with molten paraffin and solidified. The solidified blocks were trimmed to small size and sectioned using microtome (Spencer U.S.A) 3 to 5-micron thickness.

5.2.3 Toluidine blue Staining:

Toluidine blue is a metachromatic stain that stains nucleus which appears blue in color. The isolated rat's brain was fixed and processed as per the tissue processing procedure, and then brain tissue slices were prepared for toluidine blue staining. The slices were incubated with 0.5% (w/v) solution of toluidine blue for 20 minutes. After being dehydrated, cleared and mounted, brain slices were examined under a light microscope. Morphological changes in hippocampal neurons in CA1, CA3 and DG area were assessed microscopically by using 40x magnifications (Sadeghi et al,
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