R B Robinson

Columbia University, New York City, New York, United States

Are you R B Robinson?

Claim your profile

Publications (50)305.67 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: We examined whether coupling of a ventricular myocyte to a non-myocyte cell expressing HCN2 could create a two-cell syncytium capable of generating sustained pacing. Three non-myocyte cell types were transfected with the mHCN2 gene and used as sources of mHCN2-induced currents. They were human mesenchymal stem cells and HEK293 cells, both of which express connexin43 (Cx43), and HeLa cells transfected with Cx43. Cell-cell coupling between heterologous pairs increased with time in co-culture, and hyperpolarization of the myocyte induced HCN2 currents, indicating current transfer from the mHCN2-expressing cell to the myocyte via gap junctions. The magnitude of the HCN2 currents recorded in myocytes increased with increasing junctional conductance. Once a critical level of electrical cell-cell coupling between myocytes and mHCN2 transfected cells was exceeded spontaneous action potentials were generated at frequencies of approximately 0.6 to 1.7 Hz (1.09 +/- 0.05 Hz). Addition of carbenoxolone (200 microM), a gap junction channel blocker, to the media stopped spontaneous activity in heterologous cell pairs. Carbenoxolone washout restored activity. Blockade of HCN2 currents by 100 microM 9-amino-1,2,3,4-tetrahydroacridine (THA) stopped spontaneous activity and subsequent washout restored it. Neither THA nor carbenoxolone affected electrically stimulated action potentials in isolated single myocytes. In summary, the inward current evoked in the genetically engineered (HCN2-expressing) cell was delivered to the cardiac myocyte via gap junctions and generated action potentials such that the cell pair could function as a pacemaker unit. This finding lays the groundwork for understanding cell-based biological pacemakers in vivo once an understanding of delivery and target cell geometry is defined.
    The Journal of Physiology 10/2009; 587(Pt 21):5211-26. · 4.38 Impact Factor
  • I S Cohen, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: The ionic basis of automaticity in the sinoatrial node and His-Purkinje system, the primary and secondary cardiac pacemaking regions, is discussed. Consideration is given to potential targets for pharmacologic or genetic therapies of rhythm disorders. An ideal target would be an ion channel that functions only during diastole, so that action potential repolarization is not affected, and one that exhibits regional differences in expression and/or function so that the primary and secondary pacemakers can be selectively targeted. The so-called pacemaker current, If, generated by the HCN gene family, best fits these criteria. The biophysical and molecular characteristics of this current are reviewed, and progress to date in developing selective pharmacologic agents targeting If and in using gene and cell-based therapies to modulate the current are reviewed.
    Handbook of experimental pharmacology 02/2006;
  • I.S. Cohen, R.B. Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: The ionic basis of automaticity in the sinoatrial node and His-Purkinje system, the primary and secondary cardiac pacemaking regions, is discussed. Consideration is given to potential targets for pharmacologic or genetic therapies of rhythm disorders. An ideal target would be an ion channel that functions only during diastole, so that action potential repolarization is not affected, and one that exhibits regional differences in expression and/or function so that the primary and secondary pacemakers can be selectively targeted. The so-called pacemaker current, If, generated by the HCN gene family, best fits these criteria. The biophysical and molecular characteristics of this current are reviewed, and progress to date in developing selective pharmacologic agents targeting If and in using gene and cell-based therapies to modulate the current are reviewed.
    01/2006: pages 41-71;
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The purpose of this study was to determine whether oligonucleotides the size of siRNA are permeable to gap junctions and whether a specific siRNA for DNA polymerase beta (pol beta) can move from one cell to another via gap junctions, thus allowing one cell to inhibit gene expression in another cell directly. To test this hypothesis, fluorescently labelled oligonucleotides (morpholinos) 12, 16 and 24 nucleotides in length were synthesized and introduced into one cell of a pair using a patch pipette. These probes moved from cell to cell through gap junctions composed of connexin 43 (Cx43). Moreover, the rate of transfer declined with increasing length of the oligonucleotide. To test whether siRNA for pol beta was permeable to gap junctions we used three cell lines: (1) NRK cells that endogenously express Cx43; (2) Mbeta16tsA cells, which express Cx32 and Cx26 but not Cx43; and (3) connexin-deficient N2A cells. NRK and Mbeta16tsA cells were each divided into two groups, one of which was stably transfected to express a small hairpin RNA (shRNA), which gives rise to siRNA that targets pol beta. These two pol beta knockdown cell lines (NRK-kcdc and Mbeta16tsA-kcdc) were co-cultured with labelled wild type, NRK-wt or Mbeta16tsA-wt cells or N2A cells. The levels of pol beta mRNA and protein were determined by semiquantitative RT-PCR and immunoblotting. Co-culture of Mbeta16tsA-kcdc cells with Mbeta16tsA-wt, N2A or NRK-wt cells had no effect on pol beta levels in these cells. Similarly, co-culture of NRK-kcdc with N2A cells had no effect on pol beta levels in the N2A cells. In contrast, co-culture of NRK-kcdc with NRK-wt cells resulted in a significant reduction in pol beta in the wt cells. The inability of Mbeta16tsA-kcdc cells to transfer siRNA is consistent with the fact that oligonucleotides of the 12 nucleotide length were not permeable to Cx32/Cx26 channels. This suggested that Cx43 but not Cx32/Cx26 channels allowed the cell-to-cell movement of the siRNA. These results support the novel hypothesis that non-hybridized and possible hybridized forms of siRNA can move between mammalian cells through connexin-specific gap junctions.
    The Journal of Physiology 11/2005; 568(Pt 2):459-68. · 4.38 Impact Factor
  • L Protas, D DiFrancesco, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: Although the neonatal sinus node beats at a faster rate than the adult, when a sodium current (I(Na)) present in the newborn is blocked, the spontaneous rate is slower in neonatal myocytes than in adult myocytes. This suggests a possible functional substitution of I(Na) by another current during development. We used ruptured [T-type calcium current (I(Ca,T))] and perforated [L-type calcium current (I(Ca,L))] patch clamps to study developmental changes in calcium currents in sinus node cells from adult and newborn rabbits. I(Ca,T) density did not differ with age, and no significant differences were found in the voltage dependence of activation or inactivation. I(Ca,L) density was lower in the adult than newborn (12.1 +/- 1.4 vs. 17.6 +/- 2.5 pA/pF, P = 0.049). However, activation and inactivation midpoints were shifted in opposite directions, reducing the potential contribution during late diastolic depolarization in the newborn (activation midpoints -17.3 +/- 0.8 and -22.3 +/- 1.4 mV in the newborn and adult, respectively, P = 0.001; inactivation midpoints -33.4 +/- 1.4 and -28.3 +/- 1.7 mV for the newborn and adult, respectively, P = 0.038). Recovery of I(Ca,L) from inactivation was also slower in the newborn. The results suggest that a smaller but more negatively activating and rapidly recovering I(Ca,L) in the adult sinus node may contribute to the enhanced impulse initiation at this age in the absence of I(Na).
    AJP Heart and Circulatory Physiology 10/2001; 281(3):H1252-9. · 3.63 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Ventricular pacemaker current (I(f)) shows distinct voltage dependence as a function of age, activating outside the physiological range in normal adult ventricle, but less negatively in neonatal ventricle. However, heterologously expressed HCN2 and HCN4, the putative molecular correlates of ventricular I(f), exhibit only a modest difference in activation voltage. We therefore prepared an adenoviral construct (AdHCN2) of HCN2, the dominant ventricular isoform at either age, and used it to infect neonatal and adult rat ventricular myocytes to investigate the role of maturation on current gating. The expressed current exhibited an 18-mV difference in activation (V(1/2) -95.9+/-1.9 in adult; -77.6+/-1.6 mV in neonate), comparable to the 22-mV difference between native I(f) in adult and neonatal cultures (V(1/2) -98.7 versus -77.0 mV). This did not result from developmental differences in basal cAMP, because saturating cAMP in the pipette caused an equivalent positive shift in both preparations. In the neonate, AdHCN2 caused a significant increase in spontaneous rate compared with control (88+/-5 versus 48+/-4 bpm). In adult, where HCN2 activates more negatively, the effect was evident only during anodal excitation, requiring significantly less stimulus energy than control (2149+/-266 versus 3140+/-279 mV. ms). Thus, ventricular maturational state influences the voltage dependence of expressed HCN2, resulting in distinct physiological impact of expressed channels in neonate and adult myocytes. The full text of this article is available at http://www.circresaha.org.
    Circulation Research 08/2001; 89(1):E8-14. · 11.86 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The HCN family of ion channel subunits underlies the currents I(f) in heart and I(h) and I(q) in the nervous system. In the present study, we demonstrate that minK-related peptide 1 (MiRP1) is a beta subunit for the HCN family. As such, it enhances protein and current expression as well as accelerating the kinetics of activation. Because MiRP1 also functions as a beta subunit for the cardiac delayed rectifier I(Kr), these results suggest that this peptide may have the unique role of regulating both the inward and outward channels that underlie cardiac pacemaker activity. The full text of this article is available at http://www.circresaha.org.
    Circulation Research 07/2001; 88(12):E84-7. · 11.86 Impact Factor
  • Source
    M Baruscotti, D DiFrancesco, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: We have reported previously that the sinoatrial node (SAN) in the newborn rabbit expresses a Na+ current (INa) with properties similar to the neuronal type-I isoform and that this current contributes to the net inward current flowing during diastolic depolarization. To characterize this current further we conducted cell-attached single-channel experiments in isolated newborn SAN myocytes. The Na+ channel was sensitive to divalent cation block and had a single-channel conductance of 25.6 pS in the absence of divalent cations. Kinetic compatibility between single-channel and previous whole-cell data was confirmed by measuring the time constant of current decay. At pacemaker potentials, time constants were of the order of tens of milliseconds. Additional experiments indicated that this slow inactivation arises because the Na+ channels expressed in the neonatal SAN tend to re-open frequently at potentials in the pacemaker range. We suggest that this is the mechanism by which a small tetrodotoxin (TTX)-sensitive current contributes to the total inward current flowing during slow diastolic depolarization in neonatal (but not adult) pacemaker myocytes.
    Pflügers Archiv - European Journal of Physiology 06/2001; 442(2):192-6. · 4.87 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: In the present study, we demonstrate that minK-related peptide 1 (MiRP1) is a b subunit for the HCN family. As such, it enhances protein and current expression as well as accelerating the kinetics of activation. Because MiRP1 also functions as a b subunit for the cardiac delayed rectifier IKr, these results suggest that this peptide may have the unique role of regulating both the inward and outward channels that underlie cardiac pacemaker activity. The full text of this article is available at http://www.circresaha.org. (Circ Res. 2001;88:e84-e87.)
    Circulation Research - CIRC RES. 01/2001; 88(12).
  • M Baruscotti, D DiFrancesco, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: Isolated newborn, but not adult, rabbit sinoatrial node (SAN) cells exhibit spontaneous activity that (unlike adult) are highly sensitive to the Na(+) current (I(Na)) blocker TTX. To investigate this TTX action on automaticity, cells were voltage clamped with ramp depolarizations mimicking the pacemaker phase of spontaneous cells (-60 to -20 mV, 35 mV/s). Ramps elicited a TTX-sensitive current in newborn (peak density 0.89 +/- 0.14 pA/pF, n = 24) but not adult (n = 5) cells. When depolarizing ramps were preceded by steplike depolarizations to mimic action potentials, ramp current decreased 54.6 +/- 8.0% (n = 3) but was not abolished. Additional experiments demonstrated that ramp current amplitude depended on the slope of the ramp and that TTX did not alter steady-state holding current at pacemaker potentials. This excluded a steady-state Na(+) window component and suggested a kinetic basis, which was investigated by measuring TTX-sensitive I(Na) during long step depolarizations. I(Na) exhibited a slow but complete inactivation time course at pacemaker voltages (tau = 33.9 +/- 3.9 ms at -50 mV), consistent with the rate-dependent ramp data. The data indicate that owing to slow inactivation of I(Na) at diastolic potentials, a small TTX-sensitive current flows during the diastolic depolarization in neonatal pacemaker myocytes.
    AJP Heart and Circulatory Physiology 12/2000; 279(5):H2303-9. · 3.63 Impact Factor
  • Source
    J Qu, I S Cohen, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: Pacemaker current (If) exists in both neonatal and adult ventricles, but activates at more negative voltages in the adult. This study uses whole-cell patch clamp to investigate the factors that may contribute to the maturational shift of If, comparing neonatal rat ventricular myocytes that were cultured for 4-6 days either alone, in co-culture with sympathetic nerves, or with neurotransmitters chronically present in culture. If recorded from nerve-muscle co-cultures had a significantly more negative and shallower activation-voltage relation than that from control muscle cultures, which was reflected in the midpoint potential (V50) and slope factor (K) of activation. This effect of innervation was prevented by the sustained presence in the culture of the alpha1-adrenergic antagonist prazosin (Pz) at 10(-7) M. In parallel experiments, myocytes treated with noradrenaline (NA) at 10(-7) M or neuropeptide Y (NPY) at 10(-7) M during culture had the same If activation as control cells, but cells treated with NA and NPY together had a significantly more negative and shallower activation curve. Maximum conductance and reversal potential were unchanged. The effect of chronic exposure to NA + NPY was prevented by the sustained presence of either Pz or the NPY Y2 selective antagonist T4-[NPY(33-36)]4 (3.5 x 10(-7) M) in the culture, indicating a requirement for both alpha1-adrenergic and NPY Y2 activation. Substituting NA with the alpha1A-adrenergic selective agonist A61603 (5(-10) x 10(-9) M), in the presence of NPY, did not alter If, suggesting the involvement of alpha1B- rather than alpha1A-adrenoceptors. Further, sequential exposure to NPY followed by NA was effective in reproducing the action of chronic simultaneous exposure to these agonists, but sequential exposure to NA followed by NPY was ineffective. The results are consistent with past studies indicating that NPY affects the functional expression of the alpha1B-adrenergic cascade and suggest that sympathetic innervation induces a negative shift of If in ventricle via a combined action at alpha1B-adrenergic and NPY Y2 receptors. This effect of innervation probably contributes to the developmental maturation of If activation.
    The Journal of Physiology 09/2000; 526 Pt 3:561-9. · 4.38 Impact Factor
  • L Protas, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: To test the hypothesis that the Ca(2+) channel blocker mibefradil slows heart rate due to inhibition of T-type Ca(2+) current in pacemaker cells, we studied effects of mibefradil on action potentials and ionic currents of isolated rabbit sinus node cells using the patch clamp technique. Mibefradil (100 nM and 1 microM) reduced spontaneous rate, decreased action potential amplitude and finally stopped impulse initiation. This action was not due to the drug effect on hyperpolarization-activated pacemaker current, but can be explained by attenuation of both T- and L-type Ca(2+) currents, which were inhibited by mibefradil almost equally (55% and 64% inhibition with 1 microM for T- and L-types, respectively).
    European Journal of Pharmacology 08/2000; 401(1):27-30. · 2.59 Impact Factor
  • R B Robinson, Q Y Liu, M R Rosen
    [Show abstract] [Hide abstract]
    ABSTRACT: In canine ventricle, alpha-adrenergic agonists prolong action potential duration (APD) without any effect on the action potential notch, suggesting that, in this species, the effect on repolarization might be independent of inhibition of I(to). The present study investigated the action of the alpha-adrenergic agonist phenylephrine on the action potential and the repolarizing currents I(to) and I(K) in isolated canine epicardial myocytes. Isolated cells from canine epicardial tissue, and Purkinje fibers, were studied with the whole cell, voltage clamp method. Phenylephrine 0.1 microM increased APD by 13% +/- 4% at 90% repolarization without affecting the notch or amplitude. Under voltage clamp, concentrations of phenylephrine as high as 10 microM had no effect on I(to) in canine epicardial myocytes. However, I(to) of isolated canine Purkinje myocytes was reduced to 69% +/- 7% of control by 1 microM phenylephrine. Further studies in canine epicardial myocytes revealed an action of phenylephrine to inhibit I(K), and in particular I(Ks). Using a voltage protocol that included a two-step repolarization to separate I(Ks) and I(Kr) tail components, the largely I(Kr) component was not significantly affected by 1 microM phenylephrine, whereas the largely I(Ks) component was reduced to 81% +/- 5% of control value. Alpha-adrenergic prolongation of repolarization in canine epicardium does not result from inhibition of I(to). Rather, it appears that reduction of I(Ks) contributes to the action of phenylephrine. The unresponsiveness of epicardial I(to) is not a general characteristic of the canine heart, because Purkinje myocyte I(to) was inhibited, suggesting regional differences in the molecular basis of I(to) and/or alpha-adrenergic signaling in the canine heart.
    Journal of Cardiovascular Electrophysiology 02/2000; 11(1):70-6. · 3.48 Impact Factor
  • L Protas, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: The developmental increase in L-type Ca current (I(Ca,L)) density in the rat ventricle is reproduced in vitro by culturing neonatal myocytes with sympathetic neurons. We tested whether this effect of sympathetic innervation results from a chronic or sustained action of neurally released neuropeptide Y (NPY). Ventricular myocytes from newborn rats were cultured in serum-free medium with or without sympathetic neurons, NPY, or NPY analogs. Ca currents were measured in single myocytes at room temperature using the perforated patch clamp. In all cell groups (control, innervated, or NPY treated), the current-voltage relation for I(Ca,L) was represented by a bell-shaped curve with maximal value near 0 mV. The current density at 0 mV normalized to that of corresponding mean control values was 1.63 +/- 0.12 and 1.52 +/- 0.16 for innervated and NPY-treated myocytes, respectively. Both groups differed significantly from control (P < 0.05). NPY analogs exhibited the following rank order of effectiveness: NPY >/= NPY-(13-36) >/= PYY > [Leu31Pro34]NPY, suggesting that the NPY effect occurs via a Y2-receptor subtype. In confirmation, chronic treatment of innervated cultures with a Y2-selective NPY antagonist prevented the innervation-dependent increase in I(Ca,L). These results indicate that sympathetic innervation contributes to the developmental increase in I(Ca,L) via neurally released NPY acting at Y2 receptors on the ventricular myocytes.
    The American journal of physiology 09/1999; 277(3 Pt 2):H940-6. · 3.28 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: HCN cation channel mRNA expression was determined in the rabbit heart and neonatal and adult rat ventricle using RNase protection assays. In the rabbit SA node, the dominant HCN transcript is HCN4, representing >81% of the total HCN message. HCN1 is also expressed, representing >18% of the total HCN mRNA. Rabbit Purkinje fibers contained almost equal amounts of HCN1 and HCN4 transcripts with low levels of HCN2, whereas rabbit ventricle contained predominantly HCN2. The SA node contained 25 times the total HCN message of Purkinje fibers and 140 times the total HCN message of ventricle. No reports of hyperpolarization-activated current (If) exist in rabbit Purkinje fibers, and we could not record If in rabbit ventricular myocytes. To investigate the possible role of isoform switching in determining the voltage dependence of If, we determined the prevalence of HCN isoforms in neonatal and adult rat ventricle. We had previously determined the threshold for activation of If to be approximately -70 mV in neonatal rat ventricle and -113 mV in adult rat ventricle. In both neonatal and adult rat ventricle, only HCN2 and HCN4 transcripts are present. The ratio of HCN2 to HCN4 is approximately 5:1 in the neonate and 13:1 in the adult. Taken together, these results suggest that different cardiac regions express different isoforms of the HCN family. The HCN1 and HCN4 isoforms are most closely associated with a depolarized threshold for If activation, whereas the HCN2 isoform is associated with a more negative activation curve.
    Circulation Research 08/1999; 85(1):e1-6. · 11.86 Impact Factor
  • L S Sun, V O Rybin, S F Steinberg, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: The cardiac alpha1-adrenergic chronotropic response changes from stimulatory to inhibitory post-natally. The mature inhibitory response is mediated by the alpha1B-adrenoceptor and a pertussis toxin sensitive G protein. In vivo and in vitro studies identify sympathetic innervation as critical for the maturation of this inhibitory response. Additional experiments in a culture model indicate the effect of innervation is dependent on neurally released neuropeptide Y. The present study establishes that the individual signaling elements in the neuropeptide Y induced alpha1-adrenergic cascade are the same as those appearing during normal in vivo development. In addition, the data demonstrate that the effect of neuropeptide Y does not result from activation of the putative cardiac Y3 neuropeptide Y receptor subtype, since it is reproduced by the peptide fragment neuropeptide Y-(13-36) but not by [Leu31, Pro34]neuropeptide Y.
    European Journal of Pharmacology 06/1998; 349(2-3):377-81. · 2.59 Impact Factor
  • Q Y Liu, M R Rosen, D McKinnon, R B Robinson
    [Show abstract] [Hide abstract]
    ABSTRACT: During postnatal development, sympathetic innervation of the heart evolves, and repolarization accelerates. Our goal in this study was to test whether sympathetic innervation modulates the ion channels that regulate repolarization. We studied action potentials and repolarizing K+ currents in epicardial myocytes from rats in which sympathetic innervation was accelerated or delayed, respectively, by subcutaneous injection of nerve growth factor (NGF) or NGF antibody (Ab) for the first 15 days of life. A placebo group was included as well. Action potential duration (APD) to 90% repolarization was greater in the Ab (158 +/- 18 ms)-treated than the NGF (106 +/- 10 ms)-treated animals (P < 0.05); the APD at 90% repolarization for the placebo group was intermediate (125 +/- 30 ms). The transient outward (Ito) and inward rectifier (IK1) K+ currents were recorded in freshly dissociated cells using the whole cell patch-clamp technique. Ito was decreased in density at potentials positive to +40 mV in Ab-treated rats when compared with rats treated with NGF (P < 0.05). In addition, the inactivation curve of Ito in Ab-treated rats was shifted 13 mV positive to that of NGF-treated rats. IK1 also decreased in the Ab-treated group compared with the NGF group in the potential ranges of -100 to -90 mV (P < 0.05). However, the channel transcript abundance (RNA) in NGF-, Ab-, or placebo-treated rat hearts did not differ. Our results suggest that sympathetic innervation contributes to the developmental differences in K+ currents and APD postnatally in the rat.
    The American journal of physiology 03/1998; 274(3 Pt 2):H915-22. · 3.28 Impact Factor
  • A Ghavami, M Baruscotti, R B Robinson, R Hen
    [Show abstract] [Hide abstract]
    ABSTRACT: The 5-HT1B receptor is expressed on nerve terminals where it inhibits neurotransmitter release. When expressed ectopically in fibroblasts, the 5-HT1B receptor inhibits adenylyl cyclase. However, in the central nervous system, the effect of this receptor on neurotransmitter release appears to be cAMP-independent. We therefore investigated alternative effector systems that might be activated by the 5-HT1B receptor. We constructed a recombinant adenovirus that allows expression of high levels of the 5-HT1B receptor in a variety of cells. We chose cardiac ventricle myocytes because they express a muscarinic-gated, inwardly rectifying K+ channel (i[KACh]). In infected ventricle cells, both 5-HT and the muscarinic receptor agonist, carbachol, elicited a similar inwardly rectifying K+ current. The currents elicited by these agonists were pertussis-toxin sensitive and were not additive. These results suggest a common signal transduction pathway for 5-HT1B and muscarinic receptors in ventricle cells.
    European Journal of Pharmacology 12/1997; 340(2-3):259-66. · 2.59 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Neonatal rat ventricular myocytes express both beta 1-and beta 2-adrenergic receptors linked to enhanced intracellular adenosine 3',5'-cyclic monophosphate (cAMP) accumulation and the modulation of contractile function. This study tests the hypothesis that muscarinic agonists act via distinct mechanisms to interfere with beta 1-and beta 2-adrenergic receptor actions. The beta 2-selective agonist zinterol (10(-7) M) elicits approximately a fourfold increase in cAMP accumulation, which is mimicked, both in magnitude and kinetics, by 10(-9) M of the mixed beta 1-receptor agonist/beta 2-receptor agonist isoproterenol. At these concentrations, isoproterenol and zinterol elicit equivalent inotropic and lusitropic (i.e., enhanced relaxation) responses. Carbachol inhibits all three responses (cAMP, inotropic, and lusitropic) elicited by isoproterenol. In contrast, carbachol does not interfere with the effect of zinterol to augment cAMP accumulation or to induce a positive inotropic response. However, carbachol inhibits the lusitropic response to zinterol via an action at an M2-muscarinic receptor linked to a pertussis toxin-sensitive pathway. Additional studies indicate that beta 2-receptor-dependent phosphorylation of troponin I and phospholamban is substantially attenuated by carbachol. We conclude that carbachol interferes with beta 1-receptor actions by reducing cAMP accumulation. In contrast, the anti-beta 2-receptor actions of carbachol are mediated by a mechanism that is distinct from inhibition of cAMP accumulation, involving an M2-muscarinic receptor coupled to a pertussis toxin-sensitive G protein, which leads to inhibition of troponin I and phospholamban phosphorylation and inhibition of the beta 2-receptor-dependent lusitropic response.
    The American journal of physiology 07/1997; 272(6 Pt 2):H2726-35. · 3.28 Impact Factor
  • E A Accili, R B Robinson, D DiFrancesco
    [Show abstract] [Hide abstract]
    ABSTRACT: The hearts in newborn mammals have greater intrinsic beating rates, rates of diastolic depolarization, and sensitivity to autonomic stimulation than those in adults. The differences could be explained partly by altered properties of the hyperpolarization-activated current (If). To test this possibility, sinoatrial node myocytes from the hearts of newborn (9- to 10-day) and adult (>30-day) rabbits were isolated, and the If was examined with the perforated-patch-clamp technique. The fully activated current-voltage relationship yielded a larger slope conductance of If in newborn SA node myocytes (0.244 +/- 0.020 vs. 0.158 +/- 0.012 pS/pF), compatible with the more rapid diastolic depolarization. Activation curves of the If had similar midactivation voltages (newborn, -66.71 +/- 1.94 mV; adult, -66.33 +/- 2.60 mV), but the slope was significantly greater in newborns (inverse slope factor: newborn, -9.57 +/- 0.35 mV; adult, -11.34 +/- 0.54 mV). No differences in shifts of the If activation curve in response to maximal concentrations of acetylcholine (newborn, -9.70 +/- 1.8 mV; adult, -12.60 +/- 2.10 mV) and isoproterenol (newborn, 6.90 +/- 2.5 mV; adult, 5.3 +/- 1.5 mV) or in the total shift in response to these agonists (newborn, 16.60 +/- 3.30 mV; adult, 18.00 +/- 1.00 mV) were observed. The greater If density and steeper voltage dependence can contribute to both the greater heart rate and the greater sensitivity of the SA node to autonomic modulation in newborn animals.
    The American journal of physiology 04/1997; 272(3 Pt 2):H1549-52. · 3.28 Impact Factor

Publication Stats

1k Citations
305.67 Total Impact Points

Institutions

  • 1986–2009
    • Columbia University
      • • Department of Pharmacology
      • • Department of Anesthesiology
      • • Department of Medicine
      New York City, New York, United States
  • 1999–2006
    • Stony Brook University
      • Department of Physiology and Biophysics
      Stony Brook, NY, United States
  • 2001
    • University of Milan
      Milano, Lombardy, Italy
  • 1997
    • Public Health Agency
      Béal Feirste, N Ireland, United Kingdom
  • 1988–1990
    • CUNY Graduate Center
      New York City, New York, United States