Chronic stress is a risk factor for many diseases that impact the brain, including Alzheimer’s Disease. Unlike acute stress, chronic stress reduces neuronal plasticity, which can lead to neuronal remodeling and suppression. This project investigates the effect of stress on the dendritic complexity of hippocampal neurons in rats, demonstrating a methodology for procuring and analyzing these neurons. The brains of the 160 rats from the Sustained Threat and Timing (STAT) experiment were frozen. The STAT experiment investigated the effect chronic variable stress had on prospective and retrospective timing in rodents. Using a cryostat, thin coronal slices of brain tissue were placed on microscopic slides. The tissue samples were then stained using the Golgi method of silver staining. Hippocampal neurons were assessed using Sholl Analysis; the dendritic complexity of these neurons was quantified. The method of using Sholl Analysis was found to be an effective process in measuring dendritic length of hippocampal neurons.

Stress activates physiological systems within the body to protect oneself against the potential harmful effects of enduring long-term stress. Past studies have shown that structures involved in timing are implicated in a number of psychological disorders and further are sensitive to stress. In this experiment, Sprague Dawley rats are trained to perform a perspective timing task and are then exposed to twice-daily chronic variable stress for 21 days. Behavioral data are collected, followed by post-mortem tissue analysis of the PFC, hippocampus, and striatum. This study aims to examine the morphological changes in key brain regions such as the hippocampus that appear to be involved in interval timing. Additionally, this study aims to confirm that dendritic complexity in the hippocampus produces consistent data using a classic Sholl analysis versus using a virtual image-stacking software, Neurostackr. The results of this study demonstrate that the expected Gaussian graph produced from a classic Sholl analysis was produced from both a long-shaft and short-shaft neuron found in the hippocampus using the virtual technology. These findings verify that a virtual image-stacking software and Sholl analysis will suffice in place of the traditional method of hand traced neurons on a transparent sheet with concentric circles to count bifurcation points. This virtual method ultimately reduces cost, improves timeliness of data collection, and eliminates some of the subjectivity of human error.