Ideal age to record PSCs from mice spinal cord without stimulation electrode

Experimental Techniques
1

Hello!

I have two queries regarding ephys from spinal cord slice,

  1. For recording of Post synaptic currents, is there any method other than stimulation electrode to record the PSCs in mice spinal cord in P0-P12 mice spinal cord ?

Also, what is the perfect age of mice to record PSCs from spinal cord slices?

** I have recorded for 1h 10 min from P4 mice spinal cord, with only 52 events without stimulation electrode.

  1. with Leica vibratome VT1200S, is there any better parameter to optimize the protocol to get more synaptic events?

** I use 0.50 mm/s, 1.00 mm ampl and 400um slice thickness.

Best regards,
Anutosh

2

Hi Anutosh,

We routinely record sEPSC and sIPSCs in the dorsal horn from this age range.

We cut sagittal slices (350-400um thick using similar vibratome) which keeps more of the cord intact, maybe this might help?

-Best,
Judy

3

Hi Judy,

Thank you for your reply.
We are doing transverse section (300-400 uM).

At P4 mice spinal cord, we received only 51 events after 1 hr 10 minute of recording with 0.01 Hz freq. and at P13 we received 53 events after 35 minute of recording with 0.02 Hz freq. (these events are very low in number, we dont use any evoked stimulation).

Do you have any suggestion or protocol to improve the events and frequencies without any stimulation electrode or evoked response?

Best regards,
Anutosh