Abstract
Acute brain slice preparation is a powerful experimental model for investigating the characteristics of synaptic function in the brain. Although brain tissue is usually cut at ice-cold temperature (CT) to facilitate slicing and avoid neuronal damage, exposure to CT causes molecular and architectural changes of synapses. To address these issues, we investigated ultrastructural and electrophysiological features of synapses in mouse acute cerebellar slices prepared at ice-cold and physiological temperature (PT). In the slices prepared at CT, we found significant spine loss and reconstruction, synaptic vesicle rearrangement and decrease in synaptic proteins, all of which were not detected in slices prepared at PT. Consistent with these structural findings, slices prepared at PT showed higher release probability. Furthermore, preparation at PT allows electrophysiological recording immediately after slicing resulting in higher detectability of long-term depression (LTD) after motor learning compared with that at CT. These results indicate substantial advantages of the slice preparation at PT for investigating synaptic functions in different physiological conditions.
Highlights
The living acute brain slice preparation has been developed and extensively used as a powerful experimental model for investigating the structural and functional characteristics of synaptic connectivity of neuronal circuits in the brain (Li and McIlwain, 1957; Yamamoto and McIlwain, 1966; Yamamoto, 1975; Takahashi, 1978; Llinás and Sugimori, 1980)
We demonstrate that long-term depression (LTD) in mouse cerebellum caused by adaptation of horizontal optokinetic response (HOKR) is better preserved in slices prepared at physiological temperature (PT) and recorded immediately after slicing than slices prepared at cold temperature (CT) and recorded after the recovery time
We first examined whether different temperatures during slicing affect electrophysiological properties of synaptic transmission in parallel fiber-Purkinje cell (PF-Purkinje cells (PCs)) synapses in mouse cerebellum using the whole-cell patch-clamp recording
Summary
The living acute brain slice preparation has been developed and extensively used as a powerful experimental model for investigating the structural and functional characteristics of synaptic connectivity of neuronal circuits in the brain (Li and McIlwain, 1957; Yamamoto and McIlwain, 1966; Yamamoto, 1975; Takahashi, 1978; Llinás and Sugimori, 1980). The slices are pre-incubated in artificial cerebrospinal fluid (ACSF) warmed at physiological temperature (PT, 35–37◦C) for up to 1 h to recover the neuronal activities prior to electrophysiological or optic recordings (Llinás and Sugimori, 1980; Bischofberger et al, 2006). Slicing at CT and subsequent incubation at 37◦C reduces the protein level of α-amino-3-hydroxy-5-methyl4-isoxazolepropionic acid-type glutamate receptors (AMPARs), which mediate synaptic transmission and plasticity, in lysates of the acute hippocampal slices of rats (Taubenfeld et al, 2002). These artificial modifications may cause disadvantages of brain slicing at CT for investigating synaptic features
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
