+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Comprehensive Method for Culturing Embryonic Dorsal Root Ganglion Neurons for Seahorse Extracellular Flux XF24 Analysis

Comprehensive Method for Culturing Embryonic Dorsal Root Ganglion Neurons for Seahorse Extracellular Flux XF24 Analysis

Frontiers in Neurology 3: 175

Changes in mitochondrial dynamics and function contribute to progression of multiple neurodegenerative diseases including peripheral neuropathies. The Seahorse Extracellular Flux XF24 analyzer provides a comprehensive assessment of the relative state of glycolytic and aerobic metabolism in live cells making this method instrumental in assessing mitochondrial function. One of the most important steps in the analysis of mitochondrial respiration using the Seahorse XF24 analyzer is plating a uniform monolayer of firmly attached cells. However, culturing of primary dorsal root ganglion (DRG) neurons is associated with multiple challenges, including their propensity to form clumps and detach from the culture plate. This could significantly interfere with proper analysis and interpretation of data. We have tested multiple cell culture parameters including coating substrates, culture medium, XF24 microplate plastics, and plating techniques in order to optimize plating conditions. Here we describe a highly reproducible method to obtain neuron-enriched monolayers of securely attached dissociated primary embryonic (E15) rat DRG neurons suitable for analysis with the Seahorse XF24 platform.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 052269622

Download citation: RISBibTeXText

PMID: 23248613

DOI: 10.3389/fneur.2012.00175

Related references

Bioenergetic analysis of intact mammalian cells using the Seahorse XF24 Extracellular Flux analyzer and a luciferase ATP assay. Methods in Molecular Biology 1105: 589-602, 2014

Human dorsal root ganglion neurons from embryonic donors extend axons into the host rat spinal cord along laminin-rich peripheral surroundings of the dorsal root transitional zone. Journal Of Neurocytology. 26(12): 811-822,., 1997

Differential contribution of the Na(+)-K(+)-2Cl(-) cotransporter NKCC1 to chloride handling in rat embryonic dorsal root ganglion neurons and motor neurons. Faseb Journal 23(4): 1168-1176, 2009

Dye coupling between rat dorsal root ganglion neurons during embryonic development. Neuroscience Letters Supplement (32): S95, 1988

Chronic compression of mouse dorsal root ganglion alters voltage-gated sodium and potassium currents in medium-sized dorsal root ganglion neurons. Journal of Neurophysiology 106(6): 3067-3072, 2012

Distinct structure-activity relations for stimulation of 45Ca uptake and for high affinity binding in cultured rat dorsal root ganglion neurons and dorsal root ganglion membranes. Molecular Brain Research. 35(1-2): 173-182, 1996

Developmentally regulated neurite outgrowth response from dorsal root ganglion neurons to heparin-binding growth-associated molecule (HB-GAM) and the expression of HB-GAM in the targets of the developing dorsal root ganglion neurites. European Journal of Neuroscience 8(8): 1658-1665, 1996

CaMKII inactivation by extracellular Ca(2+) depletion in dorsal root ganglion neurons. Cell Calcium 39(5): 445-454, 2006

Immunohistochemical demonstration of the calcium channel alpha 2 subunit in the chicken dorsal root ganglion and spinal cord: A special reference to colocalization with calbindin-D28k in dorsal root ganglion neurons. Neuroscience Research 59(3): 304-308, 2007

Expressing Constitutively Active Rheb in Adult Dorsal Root Ganglion Neurons Enhances the Integration of Sensory Axons that Regenerate Across a Chondroitinase-Treated Dorsal Root Entry Zone Following Dorsal Root Crush. Frontiers in Molecular Neuroscience 9: 49, 2016

Activin type II receptors in embryonic dorsal root ganglion neurons of the chicken. Journal of Neurobiology 47(2): 93-108, 2001

Morphological observation on dorsal root ganglion neurons of embryonic chicken in vitro. Hua Xi Yi Ke Da Xue Xue Bao 31(2): 204-6, 216, 2003

Dye coupling between rat dorsal root ganglion neurons during embryonic development an in vitro study. Journal of Physiology (Cambridge) 426: 122P, 1990

Local application of interleukin-6 to the dorsal root ganglion induces tumor necrosis factor-α in the dorsal root ganglion and results in apoptosis of the dorsal root ganglion cells. Spine 36(12): 926-932, 2012

Action potentials of embryonic dorsal root ganglion neurons in xenopus laevis tadpoles. Journal of Physiology (Cambridge) 283: 585-604, 1978