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Depressed HCN4 function in the type 2 diabetic sinoatrial node

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Abstract

Diabetic patients often have impaired heart rate (HR) control. HR is regulated both intrinsically within the sinoatrial node (SAN) and via neuronal input. Previously, we found lower ex vivo HR in type 2 diabetic rat hearts, suggesting impaired HR generation within the SAN. The major driver of pacemaking within the SAN is the activity of hyperpolarisation-activated cyclic nucleotide-gated 4 (HCN(4)) channels. This study aimed to investigate whether the lower intrinsic HR in the type 2 diabetic heart is due to changes in HCN4 function, protein expression and/ or distribution. The intrinsic HR response to HCN4 blockade was determined in isolated Langendorff-perfused hearts of Zucker type 2 Diabetic Fatty (ZDF) rats (DM) and their non-diabetic ZDF littermates (nDM). HCN4 protein expression and membrane localisation were determined using western blot and immunofluorescence, respectively. We found that the intrinsic HR was lower in DM compared to nDM hearts. The change in intrinsic HR in response to HCN4 blockade with ivabradine was diminished in DM hearts, which normalised the intrinsic HR between the groups. HCN4 protein expression was decreased in the SAN of DM compared to nDM controls with no change in the fraction of HCN4 localised to the membrane of SAN cardiomyocytes. The lower intrinsic HR in DM is likely due to decreased HCN4 expression and depressed HCN4 function. Our study provides a novel understanding into the intrinsic mechanisms underlying altered HR control in type 2 diabetes.

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Data availability

The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

cAMP:

Cyclic adenosine monophosphate

DM:

Type 2 diabetic ZDF

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HCN4:

Hyperpolarisation-activated cyclic nucleotide-gated 4

HR:

Heart rate

If :

Funny current

LV:

Left ventricle

nDM:

Non-diabetic ZDF

RA:

Right atria

SAN:

Sinoatrial node

WGA:

Wheat germ agglutinin

ZDF:

Zucker Diabetic Fatty

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Acknowledgements

The authors wish to thank Dr Halina Dobrzynski (University of Manchester, UK) and Dr Sunil Jit Logantha (University of Liverpool, UK) for training SP to identify and isolate the sinoatrial node.

Funding

This work was supported by Heart Foundation of New Zealand Grant [#1836] and Health Research Council of New Zealand Grant [18/232] to PPJ, a Heart Foundation of New Zealand Research Fellowship to MLM [#1784] and CTB [#1783], and Project Grant [#1771] to CTB.

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Authors and Affiliations

  1. Department of Physiology, School of Biomedical Sciences, and HeartOtago, University of Otago, Dunedin, Otago, New Zealand

    Sajida Parveen, Paddy H. S. Cheah, Luke P. I. Worthington, Michelle L. Munro, Regis R. Lamberts & Peter P. Jones

  2. Department of Medicine, School of Medicine and HeartOtago, University of Otago, Dunedin, Otago, New Zealand

    Roseanna A. Smither

  3. Department of Physiology, Faculty of Medical and Health Sciences, Manaaki Mānawa Centre for Heart Research, University of Auckland, Auckland, New Zealand

    Carol T. Bussey

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  1. Sajida Parveen
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Contributions

PPJ and RRL conceived the project, secured funding, designed the experiments, reviewed the data and reviewed and edited the manuscript. SP performed the echocardiography, Langendorff and immunofluorescent experiments, collected and analysed these data and drafted the manuscript. PHSC assisted with the Langendorff and immunofluorescent experiments. CTB, LPIW and RAS performed the western blotting, analysis and interpretation. MLM oversaw the confocal imaging and image analysis and interpretation. All authors reviewed the manuscript.

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Correspondence to Peter P. Jones.

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All research was approved by the University of Otago Ethics Committee and complied with the New Zealand Animal Welfare Act 1999 and University of Otago guidelines.

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Parveen, S., Cheah, P.H.S., Worthington, L.P.I. et al. Depressed HCN4 function in the type 2 diabetic sinoatrial node. Mol Cell Biochem 478, 1825–1833 (2023). https://doi.org/10.1007/s11010-022-04635-6

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