Preparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid Pathology

Abstract

The rodent hippocampal slice preparation is perhaps the most broadly used tool for investigating mammalian synaptic function and plasticity. The hippocampus can be extracted quickly and easily from rats and mice and slices remain viable for hours in oxygenated artificial cerebrospinal fluid. Moreover, basic electrophysisologic techniques are easily applied to the investigation of synaptic function in hippocampal slices and have provided some of the best biomarkers for cognitive impairments. The hippocampal slice is especially popular for the study of synaptic plasticity mechanisms involved in learning and memory. Changes in the induction of long-term potentiation and depression (LTP and LTD) of synaptic efficacy in hippocampal slices (or lack thereof) are frequently used to describe the neurologic phenotype of cognitively-impaired animals and/ or to evaluate the mechanism of action of nootropic compounds. This article outlines the procedures we use for preparing hippocampal slices from rats and transgenic mice for the study of synaptic alterations associated with brain aging and Alzheimer's disease (AD) [1][2][3] . Use of aged rats and AD model mice can present a unique set of challenges to researchers accustomed to using younger rats and/or mice in their research. Aged rats have thicker skulls and tougher connective tissue than younger rats and mice, which can delay brain extraction and/or dissection and consequently negate or exaggerate real age-differences in synaptic function and plasticity. Aging and amyloid pathology may also exacerbate hippocampal damage sustained during the dissection procedure, again complicating any inferences drawn from physiologic assessment. Here, we discuss the steps taken during the dissection procedure to minimize these problems. Examples of synaptic responses acquired in "healthy" and "unhealthy" slices from rats and mice are provided, as well as representative synaptic plasticity experiments. The possible impact of other methodological factors on synaptic function in these animal models (e.g. recording solution components, stimulation parameters) are also discussed. While the focus of this article is on the use of aged rats and transgenic mice, novices to slice physiology should find enough detail here to get started on their own studies, using a variety of rodent models. Video LinkThe video component of this article can be found at https://www.jove.com/video/2330/ Protocol Preparing Ice-cold Oxygenated Artificial Cerebrospinal Fluid (ACSF)1. Prepare 2 L of "Ca 2+ -free" ACSF. In a 2L Erlenmeyer flask, add approximately 1.5 L of sterile or double-distilled H 2 O, and begin stirring vigorously on a stir plate. Add the following ACSF components (in mM): 124 NaCl, 2 KCl, 1.25 KH 2 PO 4 , 2 MgSO 4 , 26 NaHCO 3 , and 10 dextrose (see Table 1). Bring to a volume of 2 L with distilled H 2 O. 2. Using an aquarium bubbler and tubing attached to a 95% O 2 /5% CO 2 air tank, oxygenate ACSF vigorously for approximately 20-30 min.Check the pH, and if necessary, adjust to 7.4 using N...

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