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The Autonomic Nervous System Regulates the Heart Rate through cAMP-PKA Dependent and Independent Coupled-Clock Pacemaker Cell Mechanisms

View Article: PubMed Central - PubMed

ABSTRACT

Sinoatrial nodal cells (SANCs) generate spontaneous action potentials (APs) that control the cardiac rate. The brain modulates SANC automaticity, via the autonomic nervous system, by stimulating membrane receptors that activate (adrenergic) or inactivate (cholinergic) adenylyl cyclase (AC). However, these opposing afferents are not simply additive. We showed that activation of adrenergic signaling increases AC-cAMP/PKA signaling, which mediates the increase in the SANC AP firing rate (i.e., positive chronotropic modulation). However, there is a limited understanding of the underlying internal pacemaker mechanisms involved in the crosstalk between cholinergic receptors and the decrease in the SANC AP firing rate (i.e., negative chronotropic modulation). We hypothesize that changes in AC-cAMP/PKA activity are crucial for mediating either decrease or increase in the AP firing rate and that the change in rate is due to both internal and membrane mechanisms. In cultured adult rabbit pacemaker cells infected with an adenovirus expressing the FRET sensor AKAR3, PKA activity and AP firing rate were tightly linked in response to either adrenergic receptor stimulation (by isoproterenol, ISO) or cholinergic stimulation (by carbachol, CCh). To identify the main molecular targets that mediate between PKA signaling and pacemaker function, we developed a mechanistic computational model. The model includes a description of autonomic-nervous receptors, post- translation signaling cascades, membrane molecules, and internal pacemaker mechanisms. Yielding results similar to those of the experiments, the model simulations faithfully reproduce the changes in AP firing rate in response to CCh or ISO or a combination of both (i.e., accentuated antagonism). Eliminating AC-cAMP-PKA signaling abolished the core effect of autonomic receptor stimulation on the AP firing rate. Specifically, disabling the phospholamban modulation of the SERCA activity resulted in a significantly reduced effect of CCh and a failure to increase the AP firing rate under ISO stimulation. Directly activating internal pacemaker mechanisms led to a similar extent of changes in the AP firing rate with respect to brain receptor stimulation. Thus, Ca2+ and cAMP/PKA-dependent phosphorylation limits the rate and magnitude of chronotropic changes in the spontaneous AP firing rate.

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Related in: MedlinePlus

Sympathovagal compensation. Compensatory effects of cholinergic receptor (ChR) stimulation by CCh to adrenergic receptor (β-AR) stimulation by ISO. (A) Trend of the sympathovagal compensation curve, i.e., the concentrations of ISO and CCh that compensate for each other's effects on the action potential (AP) firing rate; (B) An example of β-AR stimulation by ISO starting at t = 900 s (10 nM) followed by ChR stimulation by CCh (88 nM) starting at t = 950 s. The AP firing rate returns to its basal rate after the addition of CCh.
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Figure 9: Sympathovagal compensation. Compensatory effects of cholinergic receptor (ChR) stimulation by CCh to adrenergic receptor (β-AR) stimulation by ISO. (A) Trend of the sympathovagal compensation curve, i.e., the concentrations of ISO and CCh that compensate for each other's effects on the action potential (AP) firing rate; (B) An example of β-AR stimulation by ISO starting at t = 900 s (10 nM) followed by ChR stimulation by CCh (88 nM) starting at t = 950 s. The AP firing rate returns to its basal rate after the addition of CCh.

Mentions: We evaluated the model's ability to reproduce some known sympathovagal effects. The compensatory effects of ChR stimulation by CCh to β-AR stimulation by ISO are shown in Figure 9. The model predicted that 88 nM of CCh would compensate for 10 nM of ISO (thus, a ratio 10:88 for rabbit SANCs). Furthermore, the model predicted a change in this ratio in response to an increase in ISO (Figure 9B). That is, relatively less CCh is needed to compensate for increased ISO treatment. Next we evaluated whether the effect of combined CCh and ISO is additive (Figure 10). Application of 20 nM of ISO increased the AP firing rate by 13% compared to the baseline (Figure 10A), while application of 80 nM CCh reduced the AP firing rate by 16% compared to the baseline (Figure 10B). However, when CCh was applied before (Figure 10C) or after ISO (Figure 10D), the net effect of both drugs was a 6% increase in the AP firing rate (i.e., not additive). Finally, we tested how and whether rapid application of CCh has a phasic effect on the AP firing rate (Figure 11). Application of short CCh pulse (see Section Methods for further details) has a phasic effect on the AP firing rate. The phasic changes in pacemaker cycle length depend on the timing of the vagal stimulation during the AP cycle. The greatest effect on AP cycle (appeared on the following beat) was observed when CCh was applied during early DD (Figure 11, see S3).


The Autonomic Nervous System Regulates the Heart Rate through cAMP-PKA Dependent and Independent Coupled-Clock Pacemaker Cell Mechanisms
Sympathovagal compensation. Compensatory effects of cholinergic receptor (ChR) stimulation by CCh to adrenergic receptor (β-AR) stimulation by ISO. (A) Trend of the sympathovagal compensation curve, i.e., the concentrations of ISO and CCh that compensate for each other's effects on the action potential (AP) firing rate; (B) An example of β-AR stimulation by ISO starting at t = 900 s (10 nM) followed by ChR stimulation by CCh (88 nM) starting at t = 950 s. The AP firing rate returns to its basal rate after the addition of CCh.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5037226&req=5

Figure 9: Sympathovagal compensation. Compensatory effects of cholinergic receptor (ChR) stimulation by CCh to adrenergic receptor (β-AR) stimulation by ISO. (A) Trend of the sympathovagal compensation curve, i.e., the concentrations of ISO and CCh that compensate for each other's effects on the action potential (AP) firing rate; (B) An example of β-AR stimulation by ISO starting at t = 900 s (10 nM) followed by ChR stimulation by CCh (88 nM) starting at t = 950 s. The AP firing rate returns to its basal rate after the addition of CCh.
Mentions: We evaluated the model's ability to reproduce some known sympathovagal effects. The compensatory effects of ChR stimulation by CCh to β-AR stimulation by ISO are shown in Figure 9. The model predicted that 88 nM of CCh would compensate for 10 nM of ISO (thus, a ratio 10:88 for rabbit SANCs). Furthermore, the model predicted a change in this ratio in response to an increase in ISO (Figure 9B). That is, relatively less CCh is needed to compensate for increased ISO treatment. Next we evaluated whether the effect of combined CCh and ISO is additive (Figure 10). Application of 20 nM of ISO increased the AP firing rate by 13% compared to the baseline (Figure 10A), while application of 80 nM CCh reduced the AP firing rate by 16% compared to the baseline (Figure 10B). However, when CCh was applied before (Figure 10C) or after ISO (Figure 10D), the net effect of both drugs was a 6% increase in the AP firing rate (i.e., not additive). Finally, we tested how and whether rapid application of CCh has a phasic effect on the AP firing rate (Figure 11). Application of short CCh pulse (see Section Methods for further details) has a phasic effect on the AP firing rate. The phasic changes in pacemaker cycle length depend on the timing of the vagal stimulation during the AP cycle. The greatest effect on AP cycle (appeared on the following beat) was observed when CCh was applied during early DD (Figure 11, see S3).

View Article: PubMed Central - PubMed

ABSTRACT

Sinoatrial nodal cells (SANCs) generate spontaneous action potentials (APs) that control the cardiac rate. The brain modulates SANC automaticity, via the autonomic nervous system, by stimulating membrane receptors that activate (adrenergic) or inactivate (cholinergic) adenylyl cyclase (AC). However, these opposing afferents are not simply additive. We showed that activation of adrenergic signaling increases AC-cAMP/PKA signaling, which mediates the increase in the SANC AP firing rate (i.e., positive chronotropic modulation). However, there is a limited understanding of the underlying internal pacemaker mechanisms involved in the crosstalk between cholinergic receptors and the decrease in the SANC AP firing rate (i.e., negative chronotropic modulation). We hypothesize that changes in AC-cAMP/PKA activity are crucial for mediating either decrease or increase in the AP firing rate and that the change in rate is due to both internal and membrane mechanisms. In cultured adult rabbit pacemaker cells infected with an adenovirus expressing the FRET sensor AKAR3, PKA activity and AP firing rate were tightly linked in response to either adrenergic receptor stimulation (by isoproterenol, ISO) or cholinergic stimulation (by carbachol, CCh). To identify the main molecular targets that mediate between PKA signaling and pacemaker function, we developed a mechanistic computational model. The model includes a description of autonomic-nervous receptors, post- translation signaling cascades, membrane molecules, and internal pacemaker mechanisms. Yielding results similar to those of the experiments, the model simulations faithfully reproduce the changes in AP firing rate in response to CCh or ISO or a combination of both (i.e., accentuated antagonism). Eliminating AC-cAMP-PKA signaling abolished the core effect of autonomic receptor stimulation on the AP firing rate. Specifically, disabling the phospholamban modulation of the SERCA activity resulted in a significantly reduced effect of CCh and a failure to increase the AP firing rate under ISO stimulation. Directly activating internal pacemaker mechanisms led to a similar extent of changes in the AP firing rate with respect to brain receptor stimulation. Thus, Ca2+ and cAMP/PKA-dependent phosphorylation limits the rate and magnitude of chronotropic changes in the spontaneous AP firing rate.

No MeSH data available.


Related in: MedlinePlus