Issue 12, 2015

Cold shock induces apoptosis of dorsal root ganglion neurons plated on infrared windows

Abstract

The chemical status of live sensory neurons is accessible with infrared microspectroscopy of appropriately prepared cells. In this paper, individual dorsal root ganglion (DRG) neurons have been prepared with two different protocols, and plated on glass cover slips, BaF2 and CaF2 substrates. The first protocol exposes the intact DRGs to 4 °C for between 20–30 minutes before dissociating individual neurons and plating 2 hours later. The second protocol maintains the neurons at 23 °C for the entire duration of the sample preparation. The visual appearance of the neurons is similar. The viability was assessed by means of trypan blue exclusion method to determine the viability of the neurons. The neurons prepared under the first protocol (cold exposure) and plated on BaF2 reveal a distinct chemical signature and chemical distribution that is different from the other sample preparations described in the paper. Importantly, results for other sample preparation methods, using various substrates and temperature protocols, when compared across the overlapping spectral bandwidth, present normal chemical distribution within the neurons. The unusual chemically specific spatial variation is dominated by a lack of protein and carbohydrates in the center of the neurons and signatures of unraveling DNA are detected. We suggest that cold shock leads to apoptosis of DRGs, followed by osmotic stress originating from ion gradients across the cell membrane leading to cell lysis.

Graphical abstract: Cold shock induces apoptosis of dorsal root ganglion neurons plated on infrared windows

Associated articles

Article information

Article type
Paper
Submitted
15 Apr 2015
Accepted
13 May 2015
First published
15 May 2015
This article is Open Access
Creative Commons BY license

Analyst, 2015,140, 4046-4056

Author version available

Cold shock induces apoptosis of dorsal root ganglion neurons plated on infrared windows

E. Aboualizadeh, E. C. Mattson, C. L. O'Hara, A. K. Smith, C. L. Stucky and C. J. Hirschmugl, Analyst, 2015, 140, 4046 DOI: 10.1039/C5AN00729A

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