|Year : 2017 | Volume
| Issue : 1 | Page : 14-17
Epigenetic programming of autonomic functions in an experimental model of apnea of prematurity
Jayasri Nanduri, Nanduri R Prabhakar
Institute for Integrative Physiology and Center for Systems Biology of O2Sensing, Biological Science Division, University of Chicago, Chicago, USA
|Date of Submission||11-Apr-2017|
|Date of Acceptance||18-Apr-2017|
|Date of Web Publication||1-Jun-2017|
Nanduri R Prabhakar
Institute for Integrative Physiology, Biological Science Division, 5841 S. Maryland Avenue, MC 5068, Room No. 711, Chicago, IL 60637
Source of Support: None, Conflict of Interest: None
Intermittent hypoxia (IH) is a hallmark manifestation of recurrent apneas, which is a major clinical problem in infants born preterm. Carotid body (CB) chemoreflex and catecholamine (CA) secretion from adrenal medullary chromaffin cells (AMCs) are two major mechanisms contributing to the maintenance of cardiorespiratory homeostasis during hypoxia. The purpose of this article is to highlight recent studies showing how neonates experiencing IH affect the CB and AMC function and their consequences in adult life. To simulate apneas, rat pups were treated with IH consisting of alternating cycles of hypoxia (1.5% O2) for 15 s and room air for 5 min, 8 h/day from ages P0–P10. Rats treated neonatal IH displayed augmented CB response to hypoxia and augmented CA secretion from AMC. Rats treated for 10 days of IH in the neonatal period were allowed to grow into adulthood. Remarkably, the effects of neonatal IH on CB and AMC persisted in the adulthood. Moreover, adult rats that were exposed to IH in neonatal period exhibited hypertension, increased incidence apnea. Analysis of the underlying molecular mechanisms revealed re-programming of the redox state by epigenetic mechanisms involving suppression of transcription of antioxidant enzyme genes by DNA hypermethylation. DNA hypomethylating agents might offer a novel therapeutic intervention to prevent early onset of cardiorespiratory morbidities caused by neonatal IH.
Keywords: Apnea of prematurity, cardiorespiratory morbidities, DNA methylation, oxidative stress
|How to cite this article:|
Nanduri J, Prabhakar NR. Epigenetic programming of autonomic functions in an experimental model of apnea of prematurity. BLDE Univ J Health Sci 2017;2:14-7
|How to cite this URL:|
Nanduri J, Prabhakar NR. Epigenetic programming of autonomic functions in an experimental model of apnea of prematurity. BLDE Univ J Health Sci [serial online] 2017 [cited 2018 Mar 23];2:14-7. Available from: http://www.bldeujournalhs.in/text.asp?2017/2/1/14/207430
Infants born preterm often exhibit recurrent apnea (brief, repeated cessation of breathing) resulting in intermittent hypoxia (IH). Cardiorespiratory responses to hypoxia depend on reflexes arising from the carotid body (CB), the primary sensory organ for monitoring arterial blood O2 levels. Carotid bodies, however, are immature at birth and show maturation of O2 sensing during the 1st week of neonatal life.,, Too little (hypoxia) or too much environmental O2(hyperoxia) during neonatal life profoundly impacts maturation of CB O2 sensing., Catecholamine (CA) secretion from adrenal medullary chromaffin cells (AMCs) is another important mechanism for maintaining cardiovascular homeostasis under hypoxia., In neonates, sympathetic innervation to the target organs is incomplete,, and hypoxia facilitates CA secretion by directly affecting the excitability of AMC., Infants with recurrent apnea exhibit autonomic dysfunctions including (a) altered sympathoadrenal function  (b) augmented ventilatory response to hypoxia, a hallmark of CB chemoreflex, and (c) cardiac arrhythmias. Recent studies on rodent models have shown that neonatal IH profoundly affects the CB O2 sensing and CA secretion from AMC. In this article, we provide a brief review of studies addressing the mechanisms underlying the effects of neonatal IH on the CB and AMC and their impact in adult life.
| Effects of Neonatal Intermittent Hypoxia on Hypoxic Sensing by the Carotid Body|| |
Carotid bodies from neonatal rat pups respond poorly to hypoxia.,, Neonatal rats exposed to chronic hypoxia exhibit reduced CB response to hypoxia., In striking contrast, rat pups exposed to IH from ages P0 to P10 (15 s of hypoxia followed by 5 min of normoxia, 9 episodes/h, 8 h/day) exhibit augmented CB response to hypoxia.,, The augmented sensory response to hypoxia could be seen in ex vivo carotid bodies, suggesting that this response is independent of circulatory changes.,, Although IH leads to a similar augmentation of the CB response to hypoxia in adult rats, there are some notable differences between the effects of IH in neonatal versus adult carotid bodies., First, the augmented hypoxic sensitivity in neonates can be seen with exposures to as little as 72 episodes of IH, whereas adult rats require as many as 720 IH episodes, suggesting that neonates are relatively more sensitive to IH than adults. Second, in IH-exposed adult rats, repetitive hypoxia leads to long-lasting increase in baseline sensory activity of the CB, a phenomenon termed as sensory long-term facilitation (sensory LTF). In striking contrast, IH is ineffective in evoking sensory LTF in neonatal carotid bodies. Third, IH has no significant effect on CB morphology in adult rats  whereas it caused hyperplasia of glomus cells in neonates. Fourth, in adult rats, the augmented CB response to hypoxia is completely reversed after the cessation of IH  whereas the effects of neonatal IH persisted into adulthood.
IH-exposed rat pups exhibit augmented hypoxic ventilatory response (HVR), a hallmark reflex response initiated by the CB., A similar increase in the HVR was also seen in preterm infants with recurrent apneas compared to infants without apneas. The enhanced HVR evoked by neonatal IH, on the one hand, may be beneficial in the initial stages as it provides adequate oxygenation in infants with apnea, thereby preventing deleterious effects of hypoxia on the central nervous system. On the other hand, if the apneas persist, instead of being beneficial, the heightened hypoxic sensitivity of the CB may lead to breathing instability and increased incidence of apneas. Indeed, neonatal rats exposed to several days of IH exhibit greater number of apneas than control rat pups.,
| Neonatal Intermittent Hypoxia Leads to Enhanced Catecholamine Secretion from Adrenal Medullary Chromaffin Cells|| |
Souvannakitti et al. examined the effects of IH on CA secretion from AMC in neonatal rats in response to hypoxia. CA secretion is monitored from dissociated chromaffin cells by carbon fiber amperometry. The number of chromaffin cells responding to hypoxia and the magnitude of CA secretion for a given level of hypoxia are greater in IH-exposed rats than the controls. The increased CA secretion by hypoxia is due to a greater number of secretory events as well as greater amount of CA released per se cretory event. IH increased both norepinephrine and epinephrine contents in neonatal adrenal medullae. In striking contrast, hypoxia-evoked CA secretion is reduced in rat pups exposed to continuous hypobaric hypoxia (0.4 ATM), suggesting that the augmented secretory response of AMC is unique to IH. Like CB, the enhanced AMC response to hypoxia is not reversed after the cessation of IH and persisted into adulthood.
| Physiological Consequences of Neonatal Intermittent Hypoxia in Adult Life|| |
Adult rats that were exposed to IH in neonatal period showed (a) augmented CB and AMC responses to hypoxia, (b) enhanced HVR, a hallmark response of the carotid chemoreflex, (c) irregular breathing, (d) greater number of apneas, and (e) hypertension and elevated plasma CA levels as compared to control rats. These findings are reminiscent of recent clinical studies showing greater incidence of sleep disordered breathing with apnea ,, and hypertension in young adults and adults, respectively who were born preterm.
| Reactive Oxygen Species: A major Cellular Mechanism Mediating the Effects of Neonatal Intermittent Hypoxia on the Carotid Body and Adrenal Medullary Chromaffin Cells|| |
The above-outlined studies demonstrate that intermittent leads to augmented hypoxic sensing by the CB and enhanced CA secretion from AMC in neonatal rats, whereas these responses were not seen with continuous exposure to hypoxia. The major difference between intermittent and continuous hypoxia is the periodic oxygenation in the former but not the latter. In this respect, IH resembles ischemia-reperfusion. It is well known that during reperfusion, there is increased generation of reactive oxygen species (ROS). The following observations demonstrate that ROS mediate the effects of neonatal IH on hypoxic sensing by the CB and AMC: (a) IH increased ROS levels in neonatal carotid bodies and adrenal medullae as evidenced by elevated malondialdehyde levels,, which represent oxidized lipids and proteins, and (b) antioxidant treatment prevented the augmented hypoxic response of the CB and AMC evoked by neonatal IH.,, Interestingly, antioxidant treatment had no effect on hyperplasia of glomus cells by IH, suggesting that IH-induced augmented hypoxic sensitivity of the neonatal CB is not secondary to increased number of glomus cells.
The elevated ROS levels by neonatal IH could be due to either increased ROS generation by pro-oxidant enzymes or decreased ROS degradation by antioxidant enzymes. The family of NADPH oxidases (Nox) constitutes one of the major sources of ROS in mammalian cells. IH-exposed neonatal rat adrenal medulla show elevated levels of Nox2 and 4 mRNAs and increase in Nox enzyme activity. On the other hand, mRNAs encoding antioxidant enzymes such as the manganese superoxide dismutase (Sod2), catalase 1, and glutathione peroxidase 1 were downregulated in IH-exposed neonatal rat carotid bodies and adrenal medullae. These observations suggest that both decreased activity of antioxidant enzymes and increased activity of Nox contribute to elevated ROS levels by neonatal IH.
| Neonatal Intermittent Hypoxia Initiates Epigenetic Programming of the Redox State|| |
Similar to cardiorespiratory changes, the increased ROS levels caused by neonatal IH were not normalized during room air recovery, rather persisted into adult life., A recent study examined the molecular mechanisms underlying the long-lasting effects of neonatal IH on ROS levels in the CB and adrenal medulla. In this study, rat pups are exposed to IH from ages P0 to P10 and then reared under room air environment (normoxia) for 40 days. Analysis of mRNAs shows increased expression of genes encoding pro-oxidant enzymes and decreased expression of genes encoding antioxidant enzymes in carotid bodies and adrenal medullae of adult rats exposed to IH in the neonatal period as compared to controls.
Epigenetic mechanisms are heritable modifications of DNA and include DNA methylation and histone modifications. Epigenetic changes result in long-term alterations in gene expression. Using the Sod2 as a model gene, Nanduri et al. showed that DNA hypermethylation contributes to neonatal IH-induced downregulation of Sod2 mRNA, protein, and the enzyme activity. These authors further identified a single CpG dinucleotide within the Sod2 gene close to the transcription initiation site that was hypermethylated in response to neonatal IH. Neonatal rats exposed to IH were treated with decitabine, an inhibitor of DNA methylation. Decitabine treatment prevented DNA hypermethylation of the Sod2 gene and restored ROS levels to control values. Molecular mechanisms mediating the persistent upregulation of pro-oxidant enzymes by neonatal IH, however, remain to be elucidated. Notwithstanding these limitations, the study by Nanduri et al. demonstrate that neonatal IH initiates epigenetic changes that lead to long-lasting increase in ROS levels in the CB and adrenal medulla.
Remarkably, decitabine treatment during the neonatal life prevented hypertension and increased apneas in adults. These observations suggest that neonatal IH predisposes to cardiorespiratory dysfunction in early adulthood involving epigenetic regulation of the redox state.
This research is supported by National Institutes of Health grants HL-PO1-90554. We gratefully acknowledge the participation of Dr. Anita Pawar, Dangjai Souvannakitti, and Ying-Jie Peng in various experiments outlined in this article.
Financial support and sponsorship
This research is supported by National Institutes of Health grants HL-PO1-90554.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Abu-Shaweesh JM, Martin RJ. Neonatal apnea: What's new? Pediatr Pulmonol 2008;43:937-44.
Blanco CE, Dawes GS, Hanson MA, McCooke HB. The response to hypoxia of arterial chemoreceptors in fetal sheep and new-born lambs. J Physiol 1984;351:25-37.
Donnelly DF. Developmental aspects of oxygen sensing by the carotid body. J Appl Physiol 2000;88:2296-301.
Carroll JL. Developmental plasticity in respiratory control. J Appl Physiol 2003;94:375-89.
Lagercrantz H, Bistoletti P. Catecholamine release in the newborn infant at birth. Pediatr Res 1977;11:889-93.
Seidler FJ, Slotkin TA. Adrenomedullary function in the neonatal rat: Responses to acute hypoxia. J Physiol 1985;358:1-16.
Thompson RJ, Jackson A, Nurse CA. Developmental loss of hypoxic chemosensitivity in rat adrenomedullary chromaffin cells. J Physiol 1997;498(Pt 2):503-10.
Takeuchi Y, Mochizuki-Oda N, Yamada H, Kurokawa K, Watanabe Y. Nonneurogenic hypoxia sensitivity in rat adrenal slices. Biochem Biophys Res Commun 2001;289:51-6.
Lagercrantz H, Sjöquist B. Deficient sympatho-adrenal activity - A cause of apnoea? Urinary excretion of catecholamines and their metabolites in preterm infants. Early Hum Dev 1980;4:405-9.
Nock ML, Difiore JM, Arko MK, Martin RJ. Relationship of the ventilatory response to hypoxia with neonatal apnea in preterm infants. J Pediatr 2004;144:291-5.
Poets CF, Samuels MP, Southall DP. Epidemiology and pathophysiology of apnoea of prematurity. Biol Neonate 1994;65:211-9.
Peng YJ, Rennison J, Prabhakar NR. Intermittent hypoxia augments carotid body and ventilatory response to hypoxia in neonatal rat pups. J Appl Physiol 2004;97:2020-5.
Donnelly DF, Doyle TP. Hypoxia-induced catecholamine release from rat carotid body, in vitro
, during maturation and following chronic hypoxia. Adv Exp Med Biol 1994;360:197-9.
Sterni LM, Bamford OS, Wasicko MJ, Carroll JL. Chronic hypoxia abolished the postnatal increase in carotid body type I cell sensitivity to hypoxia. Am J Physiol 1999;277(3 Pt 1):L645-52.
Pawar A, Peng YJ, Jacono FJ, Prabhakar NR. Comparative analysis of neonatal and adult rat carotid body responses to chronic intermittent hypoxia. J Appl Physiol 2008;104:1287-94.
Pawar A, Nanduri J, Yuan G, Khan SA, Wang N, Kumar GK, et al.
Reactive oxygen species-dependent endothelin signaling is required for augmented hypoxic sensory response of the neonatal carotid body by intermittent hypoxia. Am J Physiol Regul Integr Comp Physiol 2009;296:R735-42.
Peng YJ, Overholt JL, Kline D, Kumar GK, Prabhakar NR. Induction of sensory long-term facilitation in the carotid body by intermittent hypoxia: Implications for recurrent apneas. Proc Natl Acad Sci U S A 2003;100:10073-8.
Julien C, Bairam A, Joseph V. Chronic intermittent hypoxia reduces ventilatory long-term facilitation and enhances apnea frequency in newborn rats. Am J Physiol Regul Integr Comp Physiol 2008;294:R1356-66.
Nanduri J, Makarenko V, Reddy VD, Yuan G, Pawar A, Wang N, et al.
Epigenetic regulation of hypoxic sensing disrupts cardiorespiratory homeostasis. Proc Natl Acad Sci U S A 2012;109:2515-20.
Souvannakitti D, Kumar GK, Fox A, Prabhakar NR. Neonatal intermittent hypoxia leads to long-lasting facilitation of acute hypoxia-evoked catecholamine secretion from rat chromaffin cells. J Neurophysiol 2009;101:2837-46.
Rosen CL, Larkin EK, Kirchner HL, Emancipator JL, Bivins SF, Surovec SA, et al.
Prevalence and risk factors for sleep-disordered breathing in 8- to 11-year-old children: Association with race and prematurity. J Pediatr 2003;142:383-9.
Paavonen EJ, Strang-Karlsson S, Räikkönen K, Heinonen K, Pesonen AK, Hovi P, et al.
Very low birth weight increases risk for sleep-disordered breathing in young adulthood: The Helsinki Study of Very Low Birth Weight Adults. Pediatrics 2007;120:778-84.
Hibbs AM, Johnson NL, Rosen CL, Kirchner HL, Martin R, Storfer-Isser A, et al.
Prenatal and neonatal risk factors for sleep disordered breathing in school-aged children born preterm. J Pediatr 2008;153:176-82.
Dalziel SR, Parag V, Rodgers A, Harding JE. Cardiovascular risk factors at age 30 following pre-term birth. Int J Epidemiol 2007;36:907-15.
Ambrosio G, Tritto I, Chiariello M. The role of oxygen free radicals in preconditioning. J Mol Cell Cardiol 1995;27:1035-9.
Souvannakitti D, Nanduri J, Yuan G, Kumar GK, Fox AP, Prabhakar NR. NADPH oxidase-dependent regulation of T-type Ca2+ channels and ryanodine receptors mediate the augmented exocytosis of catecholamines from intermittent hypoxia-treated neonatal rat chromaffin cells. J Neurosci 2010;30:10763-72.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.
Bedard K, Krause KH. The NOX family of ROS-generating NADPH oxidases: Physiology and pathophysiology. Physiol Rev 2007;87:245-313.
Feinberg AP. Phenotypic plasticity and the epigenetics of human disease. Nature 2007;447:433-40.