Spinal Cord Dissection and Electrophysiology

Dissection of spinal cords and recoding were done as described^{51 (link),52 (link)}. Briefly, after perfusion, spinal cords from P6 newborn pups were removed and hemisectioned in artificial cerebrospinal fluid (aCSF) containing: NaCl (127 mM), KCl (1.9 mM), KH₂PO₄ (1.2 mM), CaCl₂ (2 mM), MgSO₄ (1 mM), NaHCO₃ (26 mM) and d-glucose (20.5 mM) with oxygenated air (95%O₂/5%CO₂). After dissection, spinal cords were moved to aCSF-containing bath. Using tightly fitting glass pipets, dorsal roots were stimulated (0.3–0.6 mA, S88X, SIU-C, Grass technologies). Extracellular potentials are recorded from ventral roots (L4 and L5) using tightly fitting glass pipets with pre-amplifier (P55, Astro-Med) and digitizer (Digidata 1440A, Molecular Devices) and a data acquisition programme (Clampex version 10, Molecular Devices). Recorded traces are averages of 20 individual stimuli applied at 0.1 Hz.

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Pooya S., Liu X., Kumar V.B., Anderson J., Imai F., Zhang W., Ciraolo G., Ratner N., Setchell K.D., Yoshida Y., Jankowski M.P, & Dasgupta B. (2014). The tumour suppressor LKB1 regulates myelination through mitochondrial metabolism. Nature communications, 5, 4993.

Publication 2014

Digidata 1440A Clampex version 10

Bath Cerebrospinal fluid Devices Dissection Dorsal roots Glucose Grass Mgso4 Nacl Nahco3 Newborn Perfusion Spinal cords Ventral roots

Corresponding Organization :

Other organizations : Cincinnati Children's Hospital Medical Center

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Variable analysis

independent variables

Stimulation of dorsal roots (0.3–0.6 mA)

dependent variables

Extracellular potentials recorded from ventral roots (L4 and L5)

control variables

Artificial cerebrospinal fluid (aCSF) composition: NaCl (127 mM), KCl (1.9 mM), KH2PO4 (1.2 mM), CaCl2 (2 mM), MgSO4 (1 mM), NaHCO3 (26 mM) and D-glucose (20.5 mM)
Oxygenation with 95% O2 and 5% CO2
Spinal cords from P6 newborn pups

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