The effects of over-expression of the FK506-binding protein FKBP12.6 on K+ currents in adult rabbit ventricular myocytes

Kettlewell, Sarah; Seidler, Tim; Smith, Godfrey L.

doi:10.1007/s00424-009-0666-y

Cited by 5 publications

(4 citation statements)

References 22 publications

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“…the number of virus particles per cell. Efficient infection of adult cardiomyocytes can generally be attained with an MOI of 100 [108–110]. Similar MOI values can be employed in neonatal cells [108, 111], although efficient infection may also be possible at lower virus concentration (MOI = 10–20 in [112, 113]).…”

Section: Gene Transfermentioning

confidence: 99%

Methods in cardiomyocyte isolation, culture, and gene transfer

Louch

Sheehan

Wolska

2011

Journal of Molecular and Cellular Cardiology

426

396

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Since techniques for cardiomyocyte isolation were first developed nearly 35 years ago, experiments on single myocytes have yielded great insight into their cellular and sub-cellular physiology. These studies have employed a broad range of techniques including electrophysiology, calcium imaging, cell mechanics, immunohistochemistry and protein biochemistry. More recently, techniques for cardiomyocyte culture have gained additional importance with the advent of gene transfer technology. While such studies require a high quality cardiomyocyte population, successful cell isolation and maintenance during culture remain challenging. In this review, we describe methods for the isolation of adult and neonatal ventricular myocytes from rat and mouse heart. This discussion outlines general principles for the beginner, but also provides detailed specific protocols and advice for common caveats. We additionally review methods for short-term myocyte culture, with particular attention given to the importance of substrate and media selection, and describe time-dependent alterations in myocyte physiology that should be anticipated. Gene transfer techniques for neonatal and adult cardiomyocytes are also reviewed, including methods for transfection (liposome, electroporation) and viral-based gene delivery.

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Section: Gene Transfermentioning

confidence: 99%

Methods in cardiomyocyte isolation, culture, and gene transfer

Louch

Sheehan

Wolska

2011

Journal of Molecular and Cellular Cardiology

426

396

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“…34 Overexpression of FKBP1A by adenovirus-transfected isolated cardiomyocytes increases the SR Ca 2+ content and decreases Ca 2+ spark frequency. 35 Calmodulin acts as part of a calcium signal transduction pathway, mediating the control of many enzymes, ion channels and other proteins through calcium binding. 36 Among the enzymes to be stimulated by the calmodulin-calcium complex are numerous protein kinases, such as myosin light-chain kinases.…”

Section: Discussionmentioning

confidence: 99%

“…FKBP1A‐deficient mice have severe cardiac defects and even die before or shortly after birth 34 . Overexpression of FKBP1A by adenovirus‐transfected isolated cardiomyocytes increases the SR Ca 2+ content and decreases Ca 2+ spark frequency 35 . Calmodulin acts as part of a calcium signal transduction pathway, mediating the control of many enzymes, ion channels and other proteins through calcium binding 36 .…”

Section: Discussionmentioning

confidence: 99%

Cardiac proteomic profiling suggests that hypertrophic and dilated cardiomyopathy share a common pathogenetic pathway of the calcium signalling pathway

Yang

Tang

et al. 2023

Eur J Clin Investigation

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ObjectiveHypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM) are classified as different diseases but have many similar pathogenic genes and clinical symptoms. Previous research has focused on mutated genes. This study was conducted to identify key molecular mechanisms and explore effective therapeutic targets.MethodsMyocardial tissue was harvested from patients with HCM (n = 3) or DCM (n = 4) during surgery. Hearts donated by healthy traffic accident victims were treated as controls (n = 4). Total proteins were extracted for liquid chromatography–tandem mass spectrometry. Differentially expressed proteins (DEPs) were annotated via GO and KEGG analyses. Selected distinguishing protein abundance was confirmed by western blotting.ResultsCompared with the control group, there were 121 and 76 DEPs in the HCM and DCM groups, respectively. GO terms for these two comparisons are associated with contraction‐related components and actin binding. Additionally, the most significantly upregulated and downregulated proteins were periostin and tropomyosin alpha‐3 chain in both comparisons. Moreover, when comparing the HCM and DCM groups, we found 60 significant DEPs, and the GO and KEGG terms are related to the calcium signalling pathway. Expression of the calcium regulation–related protein peptidyl‐prolyl cis‐trans isomerase (FKBP1A) was significantly upregulated in multiple samples.ConclusionHCM and DCM have many mutual pathogenetic pathways. Calcium ion‐related processes are among the most significant factors affecting disease development. For HCM and DCM, research on regulating linchpin protein expression or interfering with key calcium‐related pathways may be more beneficial than genetic research.

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“…Calcium-tolerant adult rabbit ventricular myocytes were isolated from the left ventricular free wall of male New Zealand White rabbits (2.8-3.2kg, 14-19wks) following perfusion with collagenase in the Langendorff mode, as described previously (Kettlewell et al, 2009). Cell Reports 31, 107697, June 9, 2020 e2…”

Section: Ventricular Myocytes and Cardiac Tissuementioning

confidence: 99%