Keywords

Cardiovascular biology, heart development, catecholamines, adrenaline, heart function, mouse model, left ventricular dysfunction, ablation, echocardiography, adrenergic cells, pnmt

Abstract

Phenylethanolamine-N-methyltransferase (Pnmt) catalyzes the conversion of noradrenaline to adrenaline and is the last enzyme in the catecholamine biosynthetic pathway. Pnmt serves as a marker for adrenergic cells, and lineage-tracing experiments have identified the embryonic heart and hindbrain region as the first sites of Pnmt expression in the mouse. Pnmt expression in the heart occurs before the adrenal glands have formed and prior to sympathetic innervation, suggesting that the heart is the first site of catecholamine production in the mouse. The function of these Pnmt+ cells in heart development remains unclear. In the present study, we test the hypothesis that (i) a genetic ablation technique utilizing a suicide reporter gene selectively destroys Pnmt cells in the mouse, and (ii) Pnmt cells are required for normal cardiovascular and neurological function. To genetically ablate adrenergic cells, we mated Pnmt-Cre mice, in which Cre-recombinase is under the transcriptional regulation of the Pnmt promoter, and a Cre -activated diphtheria toxin A (DTA) mouse strain (ROSA26-eGFP-DTA), thereby causing activation of the toxic allele (DTA) in Pnmt-expressing (adrenergic) cells resulting in selective "suicide" of these cells in approximately half of the offspring. The other half serve as controls because they do not have the ROSA26-eGFP-DTA construct. In the Pnmt+/Cre; R26+/DTA offspring, we achieve a dramatic reduction in Pnmt transcript and Pnmt immunoreactive area in the adrenal glands. Furthermore, we show that loss of Pnmt cells results in severe left ventricular dysfunction that progressively worsens with age. These mice exhibit severely reduced cardiac output and ejection fraction due to decreased LV contractility and bradycardia at rest. Surprisingly, these mice appear to have a normal stress response, as heart rate and ejection fraction increased to a similar extent compared to controls. In addition to baseline cardiac dysfunction, these mice fail to gain body weight in a normal manner and display gross neurological dysfunction, including muscular weakness, abnormal gaiting, and altered tail suspension reflex, an indicator of neurological function. This work demonstrates that selective Pnmt cell destruction leads to severe left ventricular dysfunction, lack of weight gain, and neurological dysfunction. This novel mouse is expected to shed insight into the role of Pnmt cells in the heart, and suggests a role for Pnmt cells in neurological regulation of feeding behavior, metabolism, and motor control.

Notes

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Graduation Date

2015

Semester

Spring

Advisor

Ebert, Steven

Degree

Master of Science (M.S.)

College

College of Medicine

Department

Molecular Biology and Microbiology

Degree Program

Biotechnology

Format

application/pdf

Identifier

CFE0006048

URL

http://purl.fcla.edu/fcla/etd/CFE0006048

Language

English

Release Date

11-15-2018

Length of Campus-only Access

3 years

Access Status

Masters Thesis (Open Access)

Subjects

Dissertations, Academic -- Medicine; Medicine -- Dissertations, Academic

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