Focal injection of Ethidium bromide as a simple model to study cognitive deficit and its improvement

Introduction: Memory and cognitive impairments are some of devastating outcomes of Multiple Sclerosis [MS] plaques in hippocampus, the gray matter part of the brain. The present study aimed to evaluate the intrahippocampal injection of Ethidium Bromide [EB] as a simple and focal model to assess cognition and gray matter demyelination

Methods: Thirty Wistar rats were divided into three groups: control group, which received saline, as solvent of EB, into the hippocampus; and two experimental groups, which received 3 microL of EB into the hippocampus, and then, were evaluated 7 and 28 days after EB injection [n=10 in each group], using a 5-day protocol of Morris Water Maze [MWM] task as well as Transmission Electron Microscopy [TEM] assay

Results: Seven days after EB injection, the behavioral study revealed a significance increase in travelled distance for platform finding in the experimental group compared to the control group. In addition, the nucleus of oligodendrocyte showed the typical clumped chromatin, probably attributed to apoptosis, and the myelin sheaths of some axons were unwrapped and disintegrated. Twenty-eight days after EB injection, the traveled distance and the time spent in target quadrant significantly decreased and increased, respectively in experimental groups compared to the control group. Also, TEM micrographs revealed a thin layer of remyelination around the axons in 28 days lesion group

Discussion: While intracerebral or intraventricular injection of EB is disseminated in different parts of the brain and can affect the other motor and sensory systems, this model is confined locally and facilitates behavioral study. Also, this project could show improvement of memory function subsequent to the physiological repair of the gray matter of the hippocampus

[ effect of reference memory on the number of astrocytes in different subfield of rat's hippocampus]

In addition to pyramidal neurons and interneurons, the hippocampus contains Astrocytes that play important roles in regulating ion flux currents, energy production, neurotransmitter release and memory. Learning needs some instrument for information storage and information maintenances mechanisms resemble to memory. The aim of this study was determination of spatial memory effect on the number of astrocytes in rat's hippocampus. In this experimental study, with usage of Morris Water Maze and Reference memory technique, we used 10 male albino wistar rats. 5 rats were in control group and 5 rats in Reference memory group. After histological preparation, the slides were stained with PTAH staining for showing the Astrocytes. The findings of this study showed significant difference in astrocytes number in CA1, CA2 and CA3 area of hippocampus between control and reference memory group. The mean and SD of astrocytes in CA1, CA2 and CA3 of reference memory group were 118.57 +/- 25.29, 58.91 +/- 23.59 and 116.6 +/- 31.14, that they are more than control group with 49 +/- 17.29 in CA1, 48.8 +/- 25.21 in CA2 and 41.95 +/- 11.22 in CA3. We concluded that the number of astrocytes increased due to spatial learning [e.g. reference memory method]

[ effect of spatial learning on the number of astrocytes in the rat's dentate gyrus]

Dentate gyrus is a part of hippocampal formation that plays an important role in memory and learning. Astrocytes are one of the important glial cells in nervous tissue that play a more active role in neuronal activity, including regulating ion flux currents, energy production, neurotransmitter release, and synaptogenesis. The aim of this study was to determine the spatial memory effect on the number of astrocytes in Rat's dentate gyrus. This experimental study, was done on 18 male Wistar Rats with using Morris water maze and Reference and Working memory methods. After spatial learning the Rat's brains was carried out and histological preparation was carried out, the slices were with PTAH method. The data analyzed, using T-test and One-way ANOVA. The results showed significant difference in astrocytes number in dentate gyrus area between Reference memory [300.57 +/- 5.98] and control [73.73 +/- 22.61] groups [P<0.05]. The difference between working memory [375.77 +/- 4.11] and control groups was significant. Comparing two groups there was a significant difference of number of astrocytes cell between the working memory and Reference memory group [P<0.05]. This study showed that spatial learning such as Reference memory and Working memory increase the number of astrocytes in dentate gyrus and this increase can be due to duration of learning