Investigation of TRPM2 cation channel activation in PTZ-induced SH-SY5Y cells by patch-clamp technique: Regulatory role of valproic acid


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Ahlatcı A., Bektaş M., Tülüce Y., Yıldızhan K.

7th International Brain Research School, Isparta, Turkey, 27 June - 03 July 2022, vol.14, no.4, pp.19

  • Publication Type: Conference Paper / Summary Text
  • Volume: 14
  • City: Isparta
  • Country: Turkey
  • Page Numbers: pp.19

Abstract

Investigation of TRPM2 cation channel activation in

PTZ-induced SH-SY5Y cells by patch-clamp

technique: Regulatory role of valproic acid

Adem AHLATCI1, Muhammet BEKTAŞ1, Yasin

TÜLÜCE2, Kenan YILDIZHAN3

1Department of Biophysics, Istanbul Faculty of

Medicine, Istanbul University, Istanbul, Türkiye.

2Department of Medical Biology, Faculty of Medicine,

Van Yuzuncu Yil University, Van, Türkiye.

3Department of Biophysics, Faculty of Medicine, Van

Yuzuncu Yil University, Van, Türkiye.

Neurotoxicity is the event that neurons are

damaged by natural or artificial toxic substances.

Pentylenetetrazole (PTZ) is a neurotoxic substance and is

used to produce experimental neurotoxicity. Transient

Receptor Potential Melastatin 2 (TRPM2) Channel is a

non-selective cation channel activated by adenosine

diphosphoribose (ADPR), calcium ion (Ca2+) and

reactive oxygen species and nitrogen species. It is

inactivated by N-(p-amylcinnamoyl) anthranilic acid

(ACA). Valproic acid (VPA) is a short-chain fatty acid in

the chemical structure of N-dipropyl acetic acid, it is used

as various anticonvulsants and mood stabilizers,

especially in psychiatric and neurological cases. Studies

have shown that valproic acid blocks voltage-gated

sodium or calcium channels (Bowden 2003), but its

effect on the TRPM2 channel in the neuron cell line has

not yet been clarified.

SH-SY5Y cells were divided into five groups;

control, control + ADPR, PTZ (30 μM for 24 hours),

VPA (1 mM for 24 hours), and PTZ + VPA (30 μM for

24 hours and 1 mM for 24 hours) (Taskiran et. al., 2021).

The patch-clamp technique was used to observe the

activation of the TRPM2 channel and the protective

effect of VPA in the neurotoxicity model created by PTZ.

In the study, ADPR was used as an agonist and ACA as

an antagonist for TRPM2 stimulation in SH-SY5Y cells

(Yıldızhan and Nazıroğlu 2020). The whole-cell mode

patch-clamp record showed us that in the ADPRstimulated

groups, the intracellular Ca2+ flow was highest

in the PTZ group compared to the other groups. While no

intracellular Ca2+ flow was observed in the VPA group,

intracellular Ca2+ flow was significantly inhibited in the

PTZ+VPA group compared to the PTZ group.

In conclusion, in the electrophysiological study

using the patch-clamp technique, we observed that VPA

has a regulatory effect on TRPM2 channel currents in

PTZ-induced SH-SY5Y cells.

Keywords: Epilepsy; TRPM2 channel: Valproic acid;

Patch-clamp.

References

Bowden CL. 2003. Valproate. Bipolar Disord. 5(3):189-202.

Taskiran AS, Ergul M, Gunes H, Ozturk A, Sahin B, Ozdemir E. 2021.

The Effects of Proton Pump Inhibitors (Pantoprazole) on

Pentylenetetrazole-Induced Epileptic Seizures in Rats and

Neurotoxicity in the SH-SY5Y Human Neuroblastoma Cell

Line. Cell Mol Neurobiol. 41(1):173-183. doi: 10.1007/s10571-

020-00956-6.

Yıldızhan K, Nazıroğlu M. 2020. Glutathione Depletion and

Parkinsonian Neurotoxin MPP+-Induced TRPM2 Channel

Activation Play Central Roles in Oxidative Cytotoxicity and

Inflammation in Microglia. Mol Neurobiol. 57(8):3508-3525.

doi: 10.1007/s12035-020-01974-7.