A major interest of our lab is to investigate the cellular mechanisms involved in the regulation of calcium cycling at the level of sarcoplasmic reticulum and its role in cardiac disease.  For these experiments, we apply conventional biochemical and molecular biology assays and digital confocal imaging. Model systems include established cell lines, primary cell cultures and transgenic animals. These techniques serve as valuable experimental tools to investigate PLN mutations, cellular protein sorting and Ca2+ cycling. 

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1. Phospholamban (PLN)  in Cardiac Muscle 

The overall objectives of this project are to provide a clearer understanding of the role of phospholamban (PLN) in the normal working of the heart, and what are the cellular mechanisms in human PLN genetic diseases that result in dilated cardiomyopathy. Further studies of the formation of the PLN–SERCA2a complex might provide further clues as to the mechanisms that underlie PLN regulation and provide new insights into ways to design chemicals that would disrupt this interaction selectively. Our aim is to provide such novel approaches to aid in the diagnosis and treatment of cardiac diseases resulting from defects in the regulation of intracellular calcium concentrations. 

 Shown, at left, is a schematic diagram of the 52 amino acids that constitute PLN, together with the phosphorylation sites of PLN (Ser-16 and Thr-17)

2. Sarcolipin (SLN) in Skeletal and Cardiac Muscle

The overall objectives of this project are to provide a clearer understanding of the role of the PLN homologue, sarcolipin, in the normal working of the heart, as well as skeletal muscle. We also investigate SLN interactions with SERCA since recent work has indicated that SLN and PLN can interact and bind with SERCA resulting in the superinhibition of SERCA. 

3. Targeted Transgenesis

In order to investigate the roles of several calcium regulatory proteins, we routinely generate mouse models with either transgenic overexpression of altered (mutant) or wildtype proteins .  We have recently adopted a targeted transgenic approach to the overexpression of our constructs, with the strategy shown above (together with Dr Alan Peterson, McGill U). The cDNAs of interest are inserted into a HPRT targeting vector and eventually insert into the HPRT-null locus of mutant ES cells. The result is a targeted insertion of one copy of our cDNA. For our ongoing studies, we design our constructs with tissue-specific promoters (such as alpha-MHC) to achieve skeletal- and cardiac-specific expression. 

Assistant Professor of Physiology, University of Toronto

Anthony Gramolini, PhD

Shown above is the gel-free, shotgun mass spectrometry, expression system employed 

Job Opportunities

From time to time, we will have many interesting projects for enthusiastic graduate students and postdoctoral fellows.

For Post-doctoral Applicants, the University of Toronto must recognize your Ph.D. title. We also require proficiency in spoken and written English, and a demonstrated command of a relevant scientific discipline. Your resume should demonstrate a proven track record in terms of productivity and long-term independent research potential, as evidenced by first-author peer-reviewed publications and public presentations. Foreigners must be eligible for employment authorization in Canada; please inquire at your local Canadian Embassy or Consulate about the details for obtaining a suitable work-permit and allow sufficient time to get this organized. You should be acquainted with the general academic requirements, cost-of-living, and available funding sources such as relevant local, national and international Fellowships.

We have stimulating projects for Graduate Students, and I recommend that you look into the Department of Physiology website for information about the programs, as you must first be accepted into the Departmental graduate program (http://www.uoftphysiology.com). Prospective undergraduate or MSc students need a strong academic standing to be competitive.

A few competitive positions are available for highly motivated Undergraduate students with strong academic track records, and strong letters of recommendation. To be considered you should familiarize yourself with our work (e.g. browse our Website and recent publications) and understand the nature of our research publications before you apply. You should become familiar as to what we do and how this relates to your own academic training and interests. You should understand the requirements and demands of academic research, and be confident in your own strengths, potential for academic research, and your ability to communicate in spoken and written English.