Publications

Scholarly Journals--Published

• Buchholz, JN., Behringer, EJ., Pottorf, WJ., Pearce, WJ., and Vanterpool, CK. "Age-dependent changes in Ca2+ homeostasis in peripheral neurones: implications for changes in function.." Aging Cell 6. (2007): 285-296. Calcium ions represent universal second messengers within neuronal cells integrating multiple cellular functions, such as release of neurotransmitters, gene expression, proliferation, excitability, and regulation of cell death or apoptotic pathways. The magnitude, duration and shape of stimulation-evoked intracellular calcium ([Ca2+]i) transients are determined by a complex interplay of mechanisms that modulate stimulation-evoked rises in [Ca2+]i that occur with normal neuronal function. Disruption of any of these mechanisms may have implications for the function and health of peripheral neurones during the aging process. This review focuses on the impact of advancing age on the overall function of peripheral adrenergic neurones and how these changes in function may be linked to age-related changes in modulation of [Ca2+]i regulation. The data in this review suggest that normal aging in peripheral autonomic neurones is a subtle process and does not always result in dramatic deterioration in their function. We present studies that support the idea that in order to maintain cell viability peripheral neurones are able to compensate for an age-related decline in the function of at least one of the neuronal calcium-buffering systems, smooth endoplasmic reticulum calcium ATPases, by increased function of other calcium-buffering systems, namely, the mitochondria and plasmalemma calcium extrusion. Increased mitochondrial calcium uptake may represent a 'weak point' in cellular compensation as this over time may contribute to cell death. In addition, we present more recent studies on [Ca2+]i regulation in the form of the modulation of release of calcium from smooth endoplasmic reticulum calcium stores. These studies suggest that the contribution of the release of calcium from smooth endoplasmic reticulum calcium stores is altered with age through a combination of altered ryanodine receptor levels and modulation of these receptors by neuronal nitric oxide containing neurones. (01/2007)
• Sandoval RJ, Injetti ER, Gerthoffer WT, Pearce WJ.. "Postnatal maturation modulates relations among cytosolic Ca2+, myosin light chain phosphorylation, and contractile tone in ovine cerebral arteries.." American Journal of Physiology 293. (2007): (in press)-. The present study tests the hypothesis that age-related changes in patterns of agonist-induced myofilament Ca(2+) sensitization involve corresponding differences in the relative contributions of thick-filament and thin-filament regulation to overall myofilament Ca(2+) sensitivity. Posterior communicating cerebral arteries from term fetal and non-pregnant adult sheep were used in the measurements of cytosolic Ca(2+), myosin light chain phosphorylation, and contractile tensions induced by varying concentrations of K(+) or serotonin (5-HT). The results were used to assess the relative contributions of the relations between cytosolic Ca(2+) and MLC phosphorylation (thick-filament reactivity) along with the relations between MLC phosphorylation and contractile tension (thin-filament reactivity), to overall myofilament Ca(2+) sensitivity. For K(+)-induced contractions, both fetal and adult arteries exhibited similar basal myofilament Ca(2+) sensitivity. Despite this similarity, thick-filament reactivity was greater in fetal arteries whereas thin-filament reactivity was greater in adult arteries. In contrast, 5-HT-induced contractions exhibited increased myofilament Ca(2+) sensitivity compared to K(+)-induced contractions for both fetal and adult cerebral arteries, and the magnitude of this effect was greater in fetal compared to adult arteries. When interpreted together with our previous studies of 5-HT-induced myofilament Ca(2+) sensitization, we attributed the present effects to agonist-enhancement of thick-filament reactivity in fetal arteries mediated by G-protein receptor activation of a PKC-independent but rho-A dependent pathway. In adult arteries, agonist stimulation enhanced thin-filament reactivity that was also mediated thru G-protein coupled activation of rho-A dependent and PKC independent mechanisms. Overall, the present data demonstrate that agonist-enhanced myofilament Ca(2+) sensitivity can be partitioned into separate thick-filament and thin-filament effects whose magnitudes are different between fetal and adult cerebral arteries. Key words: myofilament calcium sensitivity, vascular smooth muscle, G-protein coupled receptors. (01/2007)
• Charles SM, Zhang L, Longo LD, Buchholz JN, Pearce WJ. . " Postnatal maturation attenuates pressure-evoked myogenic tone and stretch-induced increases in Ca2+ in rat cerebral arteries.." American Journal of Physiology 293. (2007): R737-R744. Although postnatal maturation potently modulates agonist-induced cerebrovascular contractility, its effects on the mechanisms mediating cerebrovascular myogenic tone remain poorly understood. Because the regulation of calcium influx and myofilament calcium sensitivity change markedly during early postnatal life, the present study tested the general hypothesis that early postnatal maturation increases the pressure sensitivity of cerebrovascular myogenic tone via age-dependent enhancement of pressure-induced calcium mobilization and myofilament calcium sensitivity. Pressure-induced myogenic tone and changes in artery wall intracellular calcium concentrations ([Ca(2+)](i)) were measured simultaneously in endothelium-denuded, fura-2-loaded middle cerebral arteries (MCA) from pup [postnatal day 14 (P14)] and adult (6-mo-old) Sprague-Dawley rats. Increases in pressure from 20 to 80 mmHg enhanced myogenic tone in MCA from both pups and adults although the normalized magnitudes of these increases were significantly greater in pup than adult MCA. At each pressure step, vascular wall [Ca(2+)](i) was also significantly greater in pup than in adult MCA. Nifedipine significantly attenuated pressure-evoked constrictions in pup MCA and essentially eliminated all responses to pressure in the adult MCA. Both pup and adult MCA exhibited pressure-dependent increases in calcium sensitivity, as estimated by changes in the ratio of pressure-induced myogenic tone to wall [Ca(2+)](i). However, there were no differences in the magnitudes of these increases between pup and adult MCA. The results support the view that regardless of postnatal age, changes in both calcium influx and myofilament calcium sensitivity contribute to the regulation of cerebral artery myogenic tone. The greater cerebral myogenic response in P14 compared with adult MCA appears to be due to greater pressure-induced increases in [Ca(2+)](i), rather than enhanced augmentation of myofilament calcium sensitivity. (01/2007)
• Pearce WJ.. "Cerebrovascular effects of ischemic preconditioning: endothelial survivin joins the fray." American Journal of Physiology 292. (2007): H2559-H2560. (01/2007)
• Sandoval RJ, Injeti ER, Williams JM, Georthoffer WT, and Pearce WJ. . "Myogenic contractility is more dependent on myofilament calcium sensitization in term fetal than adult ovine cerebral arteries.." American Journal of Physiology 293. (2007): H548-H556. Regulation of cytosolic calcium and myofilament calcium sensitivity varies considerably with postnatal age in cerebral arteries. Because these mechanisms also govern myogenic tone, the present study used graded stretch to examine the hypothesis that myogenic tone is less dependent on calcium influx and more dependent on myofilament calcium sensitization in term fetal compared with adult cerebral arteries. Term fetal and adult posterior communicating cerebral arteries exhibited similar myogenic responses, with peak tensions averaging 24 and 26% of maximum contractile force produced in any given tissue in response to an isotonic Krebs buffer containing 122 mM K(+) (K(max)) at optimum stretch ratios (working diameter/unstressed diameter) of 2.19 and 2.23, respectively. Graded stretch increased cytosolic Ca(2+) concentration at stretch ratios >2.0 in adult arteries, but increased Ca(2+) concentration only at stretch ratios >2.3 in fetal arteries. In permeabilized arteries, myogenic tone peaked at a stretch ratio of 2.1 in both fetal and adult arteries. The fetal %K(max) values at peak myogenic tone were not significantly different at either pCa 7.0 (23%) or pCa 5.5 (25%) but were significantly less at pCa 8.0 (8.4 +/- 2.3%). Conversely, adult %K(max) values at peak myogenic tone were significantly less at both pCa 8.0 (10.4 +/- 1.8%) and pCa 7.0 (16%) than at pCa 5.5 (27%). The maximal extents of stretch-induced increases in myosin light chain phosphorylation in intact fetal (20%) and adult (17%) arteries were similar. The data demonstrate that the cerebrovascular myogenic response is highly conserved during postnatal maturation but is mediated differently in fetal and adult cerebral arteries. (01/2007)
• Nishida, N., Blood, A.B., Hunter, C.J., Bragg, S., Williams, J., Pearce, W.J., Power, G.G.. "Role of prostanoids in the regulation of cerebral blood flow during normoxia and hypoxia in the fetal sheep." Pediatric Research 60. (2006): 524-529. (06/2006)
• Angeles, D.M., Pearce, W.J.. "Effect of dopamine on vascular reactivity in near-term lamb carotids: role of the endothelium." Biological Research for Nursing 8. (2006): 97-103. (06/2006)
• Pearce, W.J., Williams, J.M., Hamade, M.W., Chang, M.M., White, C.R.. "Chronic Hypoxia Modulates Endothelium-Dependent Vasorelaxation Through Multiple Independent Mechanisms in Ovine Cranial Arteries." Advances in Experimental Medicine and Biology 578. (2006): 873-92. (06/2006)
• Lin, M.T., Hessinger, D.A., Pearce, W.J., Longo, L.D.. "Modulation of BK channel calcium affinity by differential phosphorylation in developing ovine basilar artery myocytes." American Journal of Physiology 291. (2006): H732-H740. (06/2006)
• Vanterpool, C.K., Vanterpool, E.A., Pearce, W.J., Buchholz, J.N. . "Advancing age alters the expression of the ryanodine receptor 3 isoform in adult rat superior cervical ganglia. ." Journal of Applied Physiology 101. (2006): 392-400. (05/2006)
• Pearce, W.J.. "Basic and Translational Neonatal Neuroscience Research: Whither goest the future of Physician-Scientists?." Journal of Perinatology 26. (2006): S23-S29. (04/2006)
• Xiao, D., Huang, X., Longo, L.D., Pearce, W.J., Zhang, L. "Regulation of baseline Ca2+ sensitivity in permeabilized uterine arteries: effect of pregnancy." American Journal of Physiology 291. (2006): H413-H420. (03/2006)
• Williams, J.M., White, C.R., Chang, M.M., Injeti, E.R., Zhang, L., Pearce, W.J.. "Chronic hypoxic decreases in soluble guanylate cyclase protein and enzyme activity are age-dependent in fetal and adult ovine carotid arteries." Journal of Applied Physiology 100. (2006): 1857-1866. (03/2006)
• Pearce, W. J.. "Hypoxic regulation of the fetal cerebral circulation." Journal of Applied Physiology 100. (2006): 731-738. (02/2006)
• Williams, J.M. and Pearce, W.J.. "Age-dependent modulation of endothelium-dependent vasodilatation by chronic hypoxia in ovine cranial arteries. ." Journal of Applied Physiology 100. (2006): 225-232. Although abundant evidence indicates that chronic hypoxia can induce pulmonary vascular remodeling, very little is known of the effects of chronic hypoxia on cerebrovascular structure and function, particularly in the fetus. Thus the present study explored the hypothesis that chronic hypoxemia also influences the size and shape of cerebrovascular smooth muscle and endothelial cells, with parallel changes in the reactivity of these cells to endothelium-dependent vasodilator stimuli. To test this hypothesis, measurements of endothelial and vascular smooth muscle cell size and density were made in silver-stained common carotid and middle cerebral arteries from term fetal and nonpregnant adult sheep maintained at an altitude of 3,820 m for 110 days. Chronic hypoxia induced an age-dependent remodeling that led to smooth muscle cells that were larger in fetal arteries but smaller in adult arteries. Chronic hypoxia also increased endothelial cell density in fetal arteries but reduced it in adult arteries. These combined effects resulted in an increased (adult carotid), decreased (adult middle cerebral), or unchanged (fetal arteries) per cell serosal volume of distribution for endothelial factors. Despite this heterogeneity, the magnitude of endothelium-dependent vasodilatation to A23187, measured in vitro, was largely preserved, although sensitivity to this relaxant was uniformly depressed. N(G)-nitro-L-arginine methyl ester, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, and endothelium denudation each independently blocked A23187-induced vasodilation without unmasking any residual vasoconstrictor effect. Indomethacin did not significantly attenuate A23187-induced relaxation except in the hypoxic adult middle cerebral, where a small contribution of prostanoids was evident. Vascular sensitivity to exogenous nitric oxide (NO) was uniformly increased by chronic hypoxia. From these results, we conclude that chronic hypoxia reduced endothelial NO release while also upregulating some component of the NO-cGMP-PKG vasodilator pathway. These offsetting effects appear to preserve endothelium-dependent vasodilation after adaptation to chronic hypoxia. (01/2006)
• Angeles, D.M., Chang, M., Leong, V., Oberg, K.C., Pearce, W.J. . "Dexamethasone alters vascular reactivity by enhancing COX-related vasodilatationo in fetal ovine carotids.." Biol Neonate 90. (2005): 1-878. (01/2005)
• Vanterpool, C.K., Pearce, W.J., Buchholz, J.N.. "Advancing age alters rapid and spontaneous refilling of caffeine sensitive calcium stores in sympathetic superior cervical ganglion cells.." Journal of Applied Physiology 99. (2005): 963-971. Intracellular calcium concentration ([Ca2+]i) release from smooth endoplasmic reticulum (SER) stores plays an important role in cell signaling. These stores are rapidly refilled via influx through voltage-gated calcium channels or spontaneously via store-operated calcium channels and subsequent pumping by SER Ca2+-ATPases. We measured [Ca2+]i transients in isolated fura 2-loaded superior cervical ganglion cells from 6-, 12-, 20-, and 24-mo-old Fischer 344 rats. For rapid refilling, [Ca2+]i transients were elicited by a 1) 5-s exposure to K+, 2) caffeine to release Ca2+ from SER stores, 3) K+ to refill SER Ca2+ stores, and 4) caffeine. The percent difference between the peak and rate of rise of the first and second caffeine-evoked [Ca2+]i transient significantly declined over the age range of 12-24 mo. To estimate spontaneous refilling, cells were depolarized for 5 s with 68 mM K+ (control), followed by a 10-s exposure to 10 mM caffeine "conditioning stimulus" to deplete [Ca2+]i stores. Caffeine was then rapidly applied for 5 s at defined intervals from 60 to 300 s. Integrated caffeine-evoked [Ca2+]i transients were measured and plotted as a percentage of the K+ response vs. time. The derivative of the refilling time curves significantly declined over the age range from 12-24 mo. Overall, these data suggest that the ability of superior cervical ganglion cells to sustain release of [Ca2+]i following rapid or spontaneous refilling declines with advancing age. Compromised ability to sustain calcium signaling may possibly alter the overall function of adrenergic neurons innervating the cerebrovasculature. (01/2005)
• Lin, M.T., Longo, L.D., Pearce, W.J., Hessinger, D.A.. "Calcium-Activated K+ (BK) Channel-Associated Phosphatase and Kinase Activities during Development." American Journal of Physiology 289. (2005): H414-H425. In ovine basilar arterial smooth muscle cells (SMCs), the fetal "big" Ca2+-activated K+ (BK) channel activity is significantly greater and has a lower Ca2+ setpoint than BK channels from adult cells. In the present study, we tested the hypothesis that these differences result from developmentally regulated phosphorylation of these channels. Using the patch-clamp technique and a novel in situ enzymological approach, we measured the rates and extents of changes in BK channel voltage activation from SMC inside-out patch preparations in response to selective activation and inhibition of channel-associated protein phosphatases and kinases (CAPAKs). We show that BK channel activity is modulated during development by differential phosphorylation and that the activities of CAPAKs change substantially during development. In particular, excised membrane patches from adult SMCs exhibited greater protein kinase A activity than those from a fetus. In contrast, fetal SMCs exhibited greater protein kinase G activity and phosphatase activity than adult SMCs. These findings extend our previous observation that the BK channel Ca2+ setpoint differs significantly in adult and fetal cerebrovascular myocytes and suggest a biochemical mechanism for this difference. In addition, these findings suggest that the functional stoichiometry of CAPAKs varies significantly during development and that such variation may be a hitherto unrecognized mechanism of ion channel regulation. (01/2005)
• Angeles, D.M., Wycliffe, N. Holshouser, B.A., Deming, D., Pearce, W.J., Sowers, L.C., Ashwal, S. . " Use of Opioids in Asphyxiated Term Neonates: Effects on Neuroimaging and Clinical Outcome." Pediatric Research 57. (2005): 873-878. Perinatal asphyxia is a common cause of neurologic morbidity in neonates who are born at term. Asphyxiated neonates are frequently treated with analgesic medications, including opioids, for pain and discomfort associated with their care. On the basis of previous laboratory studies suggesting that opioids may have neuroprotective effects, we conducted a retrospective review of medical records of 52 neonates who were admitted to our neonatal intensive care unit between 1995 and 2002 and had undergone magnetic resonance imaging (MRI) of the brain. Our review revealed that 33% of neonates received morphine or fentanyl. The neonates who received opioids also had experienced hypoxic/ischemic insults of greater magnitude as suggested by higher plasma lactate levels and lower 5-min Apgar scores. It is interesting that the MRI studies of neonates who were treated with opioids during the first week of life demonstrated significantly less brain injury in all regions studied. More important, follow-up studies of a subgroup of opioid-treated neonates whose MRI scans were obtained in the second postnatal week had better long-term neurologic outcomes. Our results suggest that the use of opioids in the first week of life after perinatal asphyxia have no significant long-term detrimental effects and may increase the brain's resistance to hypoxic-ischemic insults. (01/2005)
• Nauli, S.M., Williams, J.M., Gerthoffer, W.T., Pearce. W.J.. "Chronic Hypoxia Modulates Relations Among Calcium, Myosin Light Chain Phosphorylation, and Force Differently in Fetal and Adult Ovine Basilar Arteries." Journal of Applied Physiology 99. (2005): 120-127. The present study tests the hypothesis that age-related differences in the contractility of cerebral arteries from hypoxic animals involve changes in myofilament Ca++ sensitivity. Basilar arteries from term fetal and non-pregnant adult sheep maintained 110 days at 3820 m were used for measurements of [Ca++]i, myosin light chain phosphorylation, and contractile tensions induced by graded concentrations of K+ or 5HT. Slopes relating [Ca++]i to tension were similar in fetal (0.83±0.07) and adult (1.02±0.08) arteries for K+-contractions, but were significantly greater for fetal (3.77±0.64) than adult (2.00±0.13) arteries for 5HT-contractions. For both K+ and 5HT- contractions, these relations were left-shifted in fetal compared to adult arteries, indicating greater Ca++ sensitivity in fetal arteries. In contrast, slopes relating [Ca++]i to %myosin phosphorylation for K+-contractions were less in fetal (0.37±0.08) than adult (0.81±0.07) arteries and the fetal curves were right-shifted. For 5HT-contractions the slope of the [Ca++]i-phosphorylation relation was similar in fetal (0.33±0.09) and adult (0.33±0.23) arteries, indicating that 5HT depressed [Ca++]i-induced myosin phosphorylation in adult arteries. For slopes relating %myosin phosphorylation to tension, fetal values (K+:1.52±0.22, 5HT:7.66±1.70) were less than adult values (K+:2.13±0.30, 5HT:8.29±2.40) for both K+ and 5HT-induced contractions, although again fetal curves were left-shifted relative to the adult. Thus, in hypoxia-acclimatized basilar arteries, a downregulated ability of Ca++ to promote myosin phosphorylation is offset by an upregulated ability of phosphorylated myosin to produce force yielding an increased fetal myofilament Ca++ sensitivity. Postnatal maturation reprioritizes the mechanisms regulating hypoxic contractility through changes in the source of activator Ca++, the pathways governing myosin light chain phosphorylation, and its interaction with actin. (01/2005)
• Longo, L.D., Pearce, W.J.. "Fetal cerebrovascular acclimatization responses to high altitude, long-term hypoxia: A model for prenatal programming of adult disease?." American Journal of Physiology 288. (2005): R16-R24. During the past several decades, many risk factors for cerebrovascular and cardiovascular disease have been identified. More recently, it has been appreciated that inadequate nutrition and/or other intrauterine factors during fetal development may play an important role in the genesis of these conditions. An additional stress factor that may ?program? the fetus for disease later in life is that of chronic hypoxia. In studies originally designed to examine the function of developing cerebral arterial function in response to long-term hypoxia (LTH), it has become clear that many cellular and subcellular changes may have important implications for later life. Here we review some of the significant alterations in fetal cerebral artery structure and function induced by high altitude (3,280 m; 12,470 ft) LTH (~110 days). LTH is associated with augmentation or upregulation of presynaptic functions including responses to perivascular (i.e., sympathetic) nerve stimulation, and structural maturational changes. In contrast, many postsynaptic functions related to the Ca2+-dependent contractile pathway tend to be downregulated, while elements of the Ca2+-independent contraction pathway are up-regulated. The results emphasize the role of high altitude, LTH in modulating many aspects of electro-mechanical and pharmaco-mechanical coupling in the developing cerebral vasculature. A complicating factor is that the regulation of cerebrovascular tone by Ca2+-dependent and Ca2+-independent pathways changes significantly as a function of maturational age. In addition to highlighting independent regulation of various elements of the signal transduction cascade, the studies demonstrate the potential for LTH to program the fetus for cerebrovascular and other disease as an adult. (01/2005)
• Vanterpool, C.K., Vanterpool, E.A., Pearce, W.J., Buchholz, J.N. . "Advancing age alters the expression of the ryanodine receptor 3 isoform in adult rat superior cervical ganglia. ." Journal of Applied Physiology . (): -.

Abstract

• (PEER REVIEWED) Sandoval, R., Injeti, E., Williams, J.M., Pearce, W.J.. " Postnatal maturation enhances calcium influx, and reduces myofilament Ca++ sensitization in myogenic responses to cerebral artery stretch.." Journal of Cerebral Blood Flow and Metabolism 27.(Supplement) (2007): BP31-3W-. (01/2007)
• (PEER REVIEWED) Williams, J.M., Pearce, W.J. . "Chronic hypoxia and postnatal maturation alter isoform expression and specific activity of protein kinase G in ovine carotid arteries." Faseb Journal 21.(Supplement) (2007): 960.27-. (01/2007)
• (PEER REVIEWED) Sandoval, R.J., Pearce, W.J.. "Maturational differences in the contributions of thick- and thin-filament reactivity to myofilament Ca2+ sensitivity in ovine cerebral arteries. ." Faseb Journal 21.(Supplement) (2007): 960.26-. (01/2007)
• (PEER REVIEWED) Charles, S., Zhang, L., Longo, L.D., Pearce, W.J., Buchholz, J.N.. "Generation of myogenic tone requires release of calcium in rat cerebral arteries.." Faseb Journal 21.(Supplement) (2007): 960.16-. (01/2007)
• (PEER REVIEWED) Buchholz, J.N., Vanterpool, C.K., Pearce, W.J., Behringer, E.J. "Advancing age alters the contribution of release of calcium from internal stores to stimulation-evoked calcium transients." Faseb Journal 21.(Supplement) (2007): 945.9-. (01/2007)
• (PEER REVIEWED) Goyal, R., Pearce, W.J., Longo, L.D., Wilson, S.M.. "Chronic hypoxia and the influence of maturation on serotonergic contractility in ovine pulmonary arteries.." Faseb Journal 21.(Supplement) (2007): 939.3-. (01/2007)
• (PEER REVIEWED) Injeti, E.R., Sandoval, R.J., Williams, J.M., Pearce, W.J. . "Postnatal maturation increases the abundance, but decreases the specific activity of MLCK in ovine carotids." Faseb Journal 21.(Supplement) (2007): 599.7-. (01/2007)
• (PEER REVIEWED) Williams, J.M., Pearce, W.J. . "Physiological modulation of protein kinase G by chronic hypoxia and maturation in adult and fetal ovine common carotid arteries." Faseb Journal 21.(Supplement) (2007): 753.6-. (01/2007)
• (PEER REVIEWED) Sandoval, R., Pearce, W.J.. "Age-dependent stretch-induced myofilament calcium sensitization in ovine cerebral arteries ." Faseb Journal 21.(Supplement) (2007): 466.6-. (01/2007)
• (PEER REVIEWED) Charles, S.M., Zhang, L., Longo, L.D., Pearce, W.J., Buchholz, J.N. "Maturation alters calcium dependent sensitivity of pressure-dependent cerebrovascular myogenic tone." Faseb Journal 21.(Supplement) (2007): 208.15-. (01/2007)
• (PEER REVIEWED) Renan Sandoval & William J Pearce. "Age-dependent stretch-induced myofilament calcium sensitization in ovine cerebral arteries ." Faseb Journal . (2006): -. (01/2006)
• (PEER REVIEWED) James M Williams & William J Pearce. "Physiological modulation of protein kinase G by chronic hypoxia and maturation in adult and fetal ovine common carotid arteries." Faseb Journal . (2006): -. (01/2006)