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Two heterozygous Cav3.2 channel mutations in a pediatric chronic pain patient: recording condition-dependent biophysical effects

  • Ivana A. Souza
  • Maria A. Gandini
  • Miranda M. Wan
  • Gerald W. ZamponiEmail author
Ion channels, receptors and transporters

Abstract

We report expression system-dependent effects of heterozygous mutations (P769L and A1059S) in the Cav3.2 CACNA1H gene identified in a pediatric patient with chronic pain and absence seizures. The mutations were introduced individually into recombinant channels and then analyzed by means of electrophysiology. When both mutants were co-expressed in tsA-201 cells, we observed a loss of channel function, with significantly smaller current densities across a wide range of voltages (−40 to +20 mV). In addition, when both mutant channels were co-expressed, the channels opened at a more depolarizing potential with a ~5-mV right shift in the half-activation potential, with no changes in half-inactivation potential and the rate of recovery from inactivation. Interestingly, when both mutants were co-expressed in the neuronal-derived CAD cells in a different extracellular milieu, the effect was remarkably different. Although not statistically significant (p < 0.07), current densities appeared augmented compared to wild-type channels and the difference in the half-activation potential was lost. This could be attributed to the replacement of extracellular sodium and potassium with tetraethylammonium chloride. Our results show that experimental conditions can be a confounding factor in the biophysical effects of T-type calcium channel mutations found in certain neurological disorders.

Keywords

Calcium channel T-type Epilepsy Pain Mutation HEK cells 

Notes

Acknowledgments

We thank the patient and her family for their cooperation with our research team. We are grateful to Lina Chen for her technical support. We thank Dr. Francois Bernier for the ExAC database search.

Compliance with ethical standards

Financial support

This work was supported by a grant from the Canadian Institutes of Health Research to GWZ. MAG is supported by a Consejo Nacional de Ciencia y Tecnologia (Mexico) postdoctoral fellowship, and MMW was supported by an Alberta Innovates—Health Solutions summer studentship.

Conflict of interest

The authors declare that they have no conflict of interest.

Consent

Informed consent was obtained from the mother for the publication of this article.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Ivana A. Souza
    • 1
  • Maria A. Gandini
    • 1
  • Miranda M. Wan
    • 1
  • Gerald W. Zamponi
    • 1
    Email author
  1. 1.Department of Physiology and Pharmacology, Alberta Children’s Hospital Research Institute, Cumming School of MedicineUniversity of CalgaryCalgaryCanada

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