Skip to main content
SpringerLink
Log in

Anatomically “Calibrated” Isolated Respiratory Networks from Newborn Rodents

  • Protocol
  • First Online:
Isolated Central Nervous System Circuits

Part of the book series: Neuromethods ((NM,volume 73))

Abstract

En bloc and transversal slice preparations from perinatal rodents are established in vitro models for studying control of breathing by neurons and neighboring glial cells in the lower brainstem. These neural “respiratory networks in the dish” show features complementary with those in recent in vivo models that enable, for example, optogenetic manipulation for studying behavioral changes in awake animals or modulation of these circuits by higher brain regions during sleep. Contrary, the in vitro models allow respiratory network analysis at the (sub)cellular level. This is currently studied using powerful analytical tools such as quantitative pharmacology and patch-clamp recording of biophysical membrane properties of respiratory interneurons and/or motoneurons (plus neighboring glia). Increasingly, these approaches are combined with fluorescence imaging of both activity and morphology of individual cells or respiratory groups. Our recent work indicates that properties of the isolated respiratory networks depend critically on both their physical dimensions and the composition of superfusates used for their study. Based on this, we recommend to use anatomically “calibrated” rhythmic en bloc and slice preparations that we have developed and to study these models in superfusate that mimics in vivo conditions as closely as possible. We show here how these preparations are generated and present examples for pharmacological and “electrophysiological imaging” analyses that revealed novel properties of neuron–glial networks involved in respiratory control.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bianchi AL, Denavit-Saubié M, Champagnat J (1995) Central control of breathing in mammals: neuronal circuitry, membrane properties, and neurotransmitters. Physiol Rev 75:1–45

    PubMed  CAS  Google Scholar 

  2. Feldman JL, Del Negro CA (2006) Looking for inspiration: new perspectives on respiratory rhythm. Nat Rev Neurosci 7:232–242

    Article  PubMed  CAS  Google Scholar 

  3. Smith JC, Abdala AP, Rybak IA, Paton JF (2009) Structural and functional architecture of respiratory networks in the mammalian brainstem. Philos Trans R Soc Lond B Biol Sci 364:2577–2587

    Article  PubMed  Google Scholar 

  4. Suzue T (1984) Respiratory rhythm generation in the in vitro brain stem-spinal cord preparation of the neonatal rat. J Physiol 354:173–183

    PubMed  CAS  Google Scholar 

  5. Ballanyi K, Onimaru H, Homma I (1999) Respiratory network function in the isolated brainstem-spinal cord of newborn rats. Prog Neurobiol 59:583–634

    Article  PubMed  CAS  Google Scholar 

  6. Ballanyi K, Onimaru H (2009) Respiratory network analysis, isolated respiratory center functions. In: Binder M, Hirokawa N, Windhorst U (eds) Encyclopedia of neuroscience. Springer, Heidelberg/New York/Tokyo, pp 3453–3458

    Chapter  Google Scholar 

  7. Smith JC, Ellenberger HH, Ballanyi K, Richter DW, Feldman JL (1991) Pre-Bötzinger complex: a brainstem region that may generate respiratory rhythm in mammals. Science 254:726–729

    Article  PubMed  CAS  Google Scholar 

  8. Molkov YI, Abdala AP, Bacak BJ, Smith JC, Paton JF, Rybak IA (2010) Late-expiratory activity: emergence and interactions with the respiratory CpG. J Neurophysiol 104:2713–2729

    Article  PubMed  Google Scholar 

  9. Smith JC, Ballanyi K, Richter DW (1992) Whole-cell patch-clamp recordings from respiratory neurons in neonatal rat brainstem in vitro. Neurosci Lett 134:153–156

    Article  PubMed  CAS  Google Scholar 

  10. Rekling JC, Champagnat J, Denavit-Saubi M (1996) Electroresponsive properties and membrane potential trajectories of three types of inspiratory neurons in the newborn mouse brain stem in vitro. J Neurophysiol 75:795–810

    PubMed  CAS  Google Scholar 

  11. Koshiya N, Smith JC (1999) Neuronal pacemaker for breathing visualized in vitro. Nature 400:360–363

    Article  PubMed  CAS  Google Scholar 

  12. Del Negro CA, Morgado-Valle C, Feldman JL (2002) Respiratory rhythm: an emergent network property? Neuron 34:821–830

    Article  PubMed  Google Scholar 

  13. Ramirez JM, Viemari JC (2005) Determinants of inspiratory activity. Respir Physiol Neurobiol 147:145–157

    Article  PubMed  CAS  Google Scholar 

  14. Doi A, Ramirez JM (2008) Neuromodulation and the orchestration of the respiratory rhythm. Respir Physiol Neurobiol 164:96–104

    Article  PubMed  CAS  Google Scholar 

  15. Ruangkittisakul A, Okada Y, Oku Y, Koshiya N, Ballanyi K (2009) Fluorescence imaging of active respiratory networks. Respir Physiol Neurobiol 168:26–38

    Article  PubMed  Google Scholar 

  16. Gaultier C, Gallego J (2008) Neural control of breathing: insights from genetic mouse models. J Appl Physiol 104:1522–1530

    Article  PubMed  CAS  Google Scholar 

  17. Gray PA (2008) Transcription factors and the genetic organization of brain stem respiratory neurons. J Appl Physiol 104:1513–1521

    Article  PubMed  Google Scholar 

  18. Ballanyi K, Ruangkittisakul A (2009) Structure-function analysis of rhythmogenic inspiratory pre-Botzinger complex networks in “calibrated” newborn rat brainstem slices. Respir Physiol Neurobiol 168:158–178

    Article  PubMed  Google Scholar 

  19. Champagnat J, Morin-Surun MP, Bouvier J, Thoby-Brisson M, Fortin G (2011) Prenatal development of central rhythm generation. Respir Physiol Neurobiol 178:146–155

    Article  PubMed  Google Scholar 

  20. Iizuka M (2001) Respiratory activity in glossopharyngeal, vagus and accessory nerves and pharyngeal constrictors in newborn rat in vitro. J Physiol 532:535–548

    Article  PubMed  CAS  Google Scholar 

  21. Iizuka M (2004) Rostrocaudal distribution of spinal respiratory motor activity in an in vitro neonatal rat preparation. Neurosci Res 50:263–269

    Article  PubMed  Google Scholar 

  22. Iizuka M (2011) Respiration-related control of abdominal motoneurons. Respir Physiol Neurobiol 179(1):80–88

    Article  PubMed  Google Scholar 

  23. Ballanyi K, Onimaru H, Keller B (2009) Respiratory network analysis, functional imaging. In: Binder M, Hirokawa N, Windhorst U (eds) Encyclopedia of neuroscience. Springer, Heidelberg/New York/Tokyo, pp 3447–3453

    Chapter  Google Scholar 

  24. Ballanyi K, Ruangkittisakul A, Onimaru H (2009) Opioids prolong and anoxia shortens delay between onset of pre-inspiratory (pFRG) and inspiratory (preBötC) network bursting in newborn rat brainstems. Eur J Physiol (Pflügers Archiv) 458:571–587

    Article  CAS  Google Scholar 

  25. Onimaru H, Homma I (2003) A novel functional neuron group for respiratory rhythm generation in the ventral medulla. J Neurosci 23:1478–1486

    PubMed  CAS  Google Scholar 

  26. Guyenet PG, Mulkey DK (2010) Retrotrapezoid nucleus and parafacial respiratory group. Respir Physiol Neurobiol 173:244–255

    Article  PubMed  CAS  Google Scholar 

  27. Ballanyi K, Panaitescu B, Ruangkittisakul A (2010) Control of breathing by nerve glue. Sci Signal 3:pe41

    Article  PubMed  CAS  Google Scholar 

  28. Taccola G, Secchia L, Ballanyi K (2007) Anoxic augmentation of lumbar respiratory bursts and block of lumbar locomotion in newborn rat brainstem-spinal cord. J Physiol 585:507–524

    Article  PubMed  CAS  Google Scholar 

  29. Giraudin A, Cabirol-Pol MJ, Simmers J, Morin D (2008) Intercostal and abdominal respiratory motoneurons in the neonatal rat spinal cord: spatiotemporal organization and responses to limb afferent stimulation. J Neurophysiol 99:2626–2640

    Article  PubMed  Google Scholar 

  30. Smith JC, Greer JJ, Liu GS, Feldman JL (1990) Neural mechanisms generating respiratory pattern in mammalian brain stem-spinal cord in vitro. I. Spatiotemporal patterns of motor and medullary neuron activity. J Neurophysiol 64:1149–1169

    PubMed  CAS  Google Scholar 

  31. Mironov SL, Langohr K, Richter DW (1999) A1 adenosine receptors modulate respiratory activity of the neonatal mouse via the cAMP-mediated signaling pathway. J Neurophysiol 81:247–255

    PubMed  CAS  Google Scholar 

  32. Tryba AK, Ramirez JM (2004) Hyperthermia modulates respiratory pacemaker bursting properties. J Neurophysiol 92:2844–2852

    Article  PubMed  Google Scholar 

  33. Ruangkittisakul A, Schwarzacher SW, Secchia L, Poon BY, Ma Y, Funk GD, Ballanyi K (2006) High sensitivity to neuromodulator-activated signaling pathways at physiological [K+] of confocally imaged respiratory center neurons in on-line-calibrated newborn rat brainstem slices. J Neurosci 26:11870–11880

    Article  PubMed  CAS  Google Scholar 

  34. Ruangkittisakul A, Secchia L, Bornes TD, Palathinkal DM, Ballanyi K (2007) Dependence on extracellular Ca2+/K+ antagonism of inspiratory centre rhythms in slices and en bloc preparations of newborn rat brainstem. J Physiol 584:489–508

    Article  PubMed  CAS  Google Scholar 

  35. Ballanyi K, Ruangkittisakul A (2009) Brain slices. In: Binder M, Hirokawa N, Windhorst U (eds) Encyclopedia of neuroscience. Springer, Heidelberg/New York/Tokyo, pp 483–490

    Chapter  Google Scholar 

  36. Hansen AJ (1985) Effect of anoxia on ion distribution in the brain. Physiol Rev 65:101–148

    PubMed  CAS  Google Scholar 

  37. Brockhaus J, Ballanyi K, Smith JC, Richter DW (1993) Microenvironment of respiratory neurons in the in vitro brainstem-spinal cord of neonatal rats. J Physiol 462:421–445

    PubMed  CAS  Google Scholar 

  38. Völker A, Ballanyi K, Richter DW (1995) Anoxic disturbance of the isolated respiratory network of neonatal rats. Exp Brain Res 103:9–19

    Article  PubMed  Google Scholar 

  39. Somjen GG (2002) Ion regulation in the brain: implications for pathophysiology. Neuroscien­tist 8:254–267

    PubMed  CAS  Google Scholar 

  40. Ruangkittisakul A, Schwarzacher SW, Secchia L, Ma Y, Bobocea N, Poon BY, Funk GD, Ballanyi K (2008) Generation of eupnea and sighs by a spatiochemically organized inspiratory network. J Neurosci 28:2447–2458

    Article  PubMed  CAS  Google Scholar 

  41. Ruangkittisakul A, Panaitescu B, Ballanyi K (2011) K+ and Ca2+ dependence of inspiratory-related rhythm in novel “calibrated” mouse brainstem slices. Respir Physiol Neurobiol 175:37–48

    Article  PubMed  CAS  Google Scholar 

  42. Panaitescu B, Ruangkittisakul A, Ballanyi K (2009) Silencing by raised extracellular Ca2+ of pre-Bötzinger complex neurons in newborn rat brainstem slices without change of membrane potential or input resistance. Neurosci Lett 456:25–29

    Article  PubMed  CAS  Google Scholar 

  43. Ruangkittisakul A, Ballanyi K (2009) Neuron-glia-imaging. In: Binder M, Hirokawa N, Windhorst U (eds) Encyclopedia of neuroscience. Springer, Heidelberg/New York/Tokyo, pp 2756–2764

    Chapter  Google Scholar 

  44. Ruangkittisakul A, Ballanyi K (2010) Methylxanthine reversal of opioid-evoked inspiratory depression via phosphodiesterase-4 blockade. Respir Physiol Neurobiol 172:94–105

    Article  PubMed  CAS  Google Scholar 

  45. Trapp S, Ballanyi K (2012) Autonomic nervous system in vitro: studying tonically active neurons controlling vagal outflow in rodent brainstem slices. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 1–59

    Google Scholar 

  46. Ballanyi K (2004) Neuromodulation of the perinatal respiratory network. Curr Neuropharmacol 2:221–243

    Article  CAS  Google Scholar 

  47. Greer JJ, Funk GD, Ballanyi K (2006) Preparing for the first breath: prenatal maturation of respiratory neural control. J Physiol 570:437–444

    Article  PubMed  CAS  Google Scholar 

  48. Ballanyi K (1999) Isolated tissues: in vitro preparations. In: Windhorst U, Johansson H (eds) Modern techniques in neuroscience research. Springer, Heidelberg, pp 307–326

    Chapter  Google Scholar 

  49. Neher E, Sakmann B (2009) Single-channel recording, 2nd edn. Springer, New York/Dordrecht/Heidelberg/London

    Google Scholar 

  50. Walz W (2007) Patch-clamp analysis: advanced techniques, 2nd edn. Humana, Totowa

    Book  Google Scholar 

  51. Onimaru H, Ballanyi K, Richter DW (1996) Calcium-dependent responses in neurons of the isolated respiratory network of newborn rats. J Physiol 491:677–695

    PubMed  CAS  Google Scholar 

  52. Onimaru H, Ballanyi K, Homma I (2003) Contribution of Ca2+-dependent conductances to membrane potential fluctuations of medullary respiratory neurons of newborn rats in vitro. J Physiol 552:727–741

    Article  PubMed  CAS  Google Scholar 

  53. Schwarzacher S, Pestean A, Günther S, Ballanyi K (2002) Serotonergic modulation of respiratory motoneurons and interneurons in ­brainstem slices of perinatal rats. Neuroscience 115:1247–1259

    Article  PubMed  CAS  Google Scholar 

  54. Moore AR, Zhou WL, Jakovcevski I, Zecevic N, Antic SD (2012) Physiological properties of human fetal cortex in vitro. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 125–158

    Google Scholar 

  55. Sanchez-Vives MV (2012) Spontaneous rhythmic activity in the adult cerebral cortex in vitro. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 263–284

    Google Scholar 

  56. Broicher T, Speckmann EJ (2012) Living human brain slices: network analysis using voltage sensitive dyes. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 285–300

    Google Scholar 

  57. Kantor C, Panaitescu B, Kuribayashi J, Ruangkittisakul A, Jovanovic I, Leung V, Lee TF, MacTavish D, Jhamandas JH, Cheung PY, Ballanyi K (2012) Spontaneous neural network oscillations in hippocampus, cortex and locus coeruleus of newborn rat and piglet brain slices. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 315–356

    Google Scholar 

  58. Biggs JE, Lu VB, Kim H, Lai A, Todd KG, Ballanyi K, Colmers WF, Smith PA (2012) Defined medium organotypic cultures of spinal cord put ‘pain in a dish’. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 405–435

    Google Scholar 

  59. Brockhaus J, Ballanyi K (1998) Synaptic inhibition in the isolated respiratory network of neonatal rats. Eur J Neurosci 10:3823–3839

    Article  PubMed  CAS  Google Scholar 

  60. Brockhaus J, Ballanyi K (2000) Anticonvulsant adenosine A1 receptor-mediated adenosine action on neuronal networks in the brainstem-spinal cord of newborn rats. Neuroscience 96:359–371

    Article  PubMed  CAS  Google Scholar 

  61. Huxtable AG, Zwicker JD, Alvares TS, Ruangkittisakul A, Fang X, Hahn LB, Posse de Chaves E, Baker GB, Ballanyi K, Funk GD (2010) Glia contribute to the purinergic modulation of inspiratory rhythm generating networks. J Neurosci 30:3947–3958

    Article  PubMed  CAS  Google Scholar 

  62. Yuste R, Konnerth A, Masters B (2006) Imaging in neuroscience and development, a laboratory manual. J Biomed Opt 11:19902

    Article  Google Scholar 

  63. Fish KN, Gonzales-Burgos G, Zaitsev AV, Lewis DA (2012) Histological characterization of physiologically determined fast spiking interneurons in slices of the primate dorsolateral prefrontal cortex. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 159–181

    Google Scholar 

  64. Nakamura TJ, Michel S, Block GD, Colwell CS (2012) Neural circuits underlying circadian oscillations in mammals: clocks in a dish. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 183–210

    Google Scholar 

  65. Stosiek C, Garaschuk O, Holthoff K, Konnerth A (2003) In vivo two-photon calcium imaging of neuronal networks. Proc Natl Acad Sci USA 100:7319–7324

    Article  PubMed  CAS  Google Scholar 

  66. Paxinos G, Halliday G, Watson C, Koutcherov Y, Wang H (2007) Atlas of the developing mouse brain. Elsevier, Amsterdam/Boston/Heidelberg

    Google Scholar 

  67. Greene EC (1935) The anatomy of the rat. Braintree Scientific, Philadelphia

    Google Scholar 

  68. Cho YR, Pawela CP, Li R, Kao D, Schulte ML, Runquist ML, Yan JG, Matloub HS, Jaradeh SS, Hudetz AG, Hyde JS (2007) Refining the sensory and motor ratunculus of the rat upper extremity using fMRI and direct nerve stimulation. Magn Reson Med 58:901–909

    Article  PubMed  Google Scholar 

  69. Mandadi S, Nakanishi ST, Han P, Humphreys J, Whela PJ (2012) The use of rodent isolated spinal cord preparations to examine motor output. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 437–464

    Google Scholar 

  70. Ballanyi K, Völker A, Richter DW (1994) Anoxia induced functional inactivation of neonatal respiratory neurons in vitro. Neuroreport 6:165–168

    Article  PubMed  CAS  Google Scholar 

  71. Ballanyi K, Lalley PM, Hoch B, Richter DW (1997) cAMP dependent reversal of opioid and prostaglandin mediated depression of the isolated respiratory network in newborn rats. J Physiol 504:127–134

    Article  PubMed  CAS  Google Scholar 

  72. Ballanyi K (2004) Protective role of neuronal KATP channels in brain hypoxia. J Exp Biol 207:3201–3212

    Article  PubMed  CAS  Google Scholar 

  73. Stachniak TS, Sudbury JR, Trudel E, Choe KY, Ciura S, Bourque CW (2012) Osmoregulatory circuits in slices and en bloc preparations of rodent hypothalamus. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 211–231

    Google Scholar 

  74. Trapp S, Lückermann M, Brooks PS, Ballanyi K (1996) Acidosis of dorsal vagal neurons in situ during spontaneous and evoked activity. J Physiol 496:695–710

    PubMed  CAS  Google Scholar 

  75. Ben-Ari Y, Gaiarsa JL, Tyzio R, Khazipov R (2007) GABA: a pioneer transmitter that excites immature neurons and generates primitive oscillations. Physiol Rev 87:1215–1284

    Article  PubMed  CAS  Google Scholar 

  76. McKay BE, Tadayonnejad R, Anderson DM, Engbers JDT, Fernandez FR, Iftinca M, Turner RW (2012) Establishing in vivo like activity in rat cerebellar cells maintained in vitro. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 233–262

    Google Scholar 

  77. Ballanyi K, Grafe P, ten Bruggencate G (1987) Ion activities and potassium uptake mechanisms of glial cells in guinea-pig olfactory cortex slices. J Physiol 382:159–174

    PubMed  CAS  Google Scholar 

  78. Konnerth A, Keller BU, Ballanyi K, Yaari Y (1990) Voltage sensitivity of NMDA-receptor mediated postsynaptic currents. Exp Brain Res 81:209–212

    Article  PubMed  CAS  Google Scholar 

  79. Ballanyi K, Branchereau P, Champagnat J, Fortin G, Velluti J (1993) Extracellular potassium, glial and neuronal potentials in the solitary complex of rat brainstem slices. Brain Res 607:99–107

    Article  PubMed  CAS  Google Scholar 

  80. Ballanyi K, Doutheil J, Brockhaus J (1996) Membrane potentials and microenvironment of rat dorsal vagal cells in vitro during energy depletion. J Physiol 495:769–784

    PubMed  CAS  Google Scholar 

  81. Ballanyi K, Kaila K (1998) Activity-evoked changes in intracellular pH. In: Kaila K, Ransom BR (eds) pH and brain function. Wiley, New York, pp 283–300

    Google Scholar 

  82. Kulik A, Haentzsch A, Lückermann M, Reichelt W, Ballanyi K (1999) Neuron-glia signaling via α1 adrenoceptor-mediated Ca2+ release in Bergmann glia in situ. J Neurosci 19:8401–8408

    PubMed  CAS  Google Scholar 

  83. Ballanyi K, Kuwana S, Völker A, Morawietz G, Richter DW (1992) Developmental changes in the hypoxia tolerance of the in vitro respiratory network of rats. Neurosci Lett 148:141–144

    Article  PubMed  CAS  Google Scholar 

  84. Ballanyi K, Völker A, Richter DW (1996) Functional relevance of anaerobic metabolism in the isolated respiratory network of newborn rats. Eur J Physiol (Pflügers Archiv) 432:741–748

    Article  CAS  Google Scholar 

  85. Voipio J, Ballanyi K (1997) Interstitial PCO2 and pH, and their role as chemostimulants in the isolated respiratory network of neonatal rats. J Physiol 499:527–542

    PubMed  CAS  Google Scholar 

  86. Ammann D (1986) Ion-selective microelectrodes. Springer, Berlin/Heidelberg/New York/Tokyo

    Google Scholar 

  87. Baumgärtl H (1987) Systematic investigations of needle electrode properties in polarographic measurements of local tissue P02. In: Ehrly AM, Hauss J, Kolde G (eds) Clinical oxygen pressure measurement. Springer, Berlin/Heidelberg/New York/London/Paris/Tokyo, pp 17–42

    Chapter  Google Scholar 

  88. Ballanyi K, Grafe P (1985) An intracellular analysis of γ-aminobutyric-acid-associated ion movements in rat sympathetic neurones. J Physiol 365:41–58

    PubMed  CAS  Google Scholar 

  89. Ballanyi K, Mückenhoff K, Bellingham MC, Okada Y, Scheid P, Richter DW (1994) Activity-related pH changes in respiratory neurones and glial cells of cats. Neuroreport 6:33–36

    Article  PubMed  CAS  Google Scholar 

  90. De Curtis M, Lilbrizzi L, Uva L, Gnatkovsky V (2012) Neuronal networks in the in vitro isolated Guinea Pig Brain. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 357–383

    Google Scholar 

  91. Day TA, Wilson RJA (2012) The study of respiratory chemoreflexes using a novel dual-perfused rodent preparation. In Isolated Central Nervous System Circuits (Ed K Ballanyi), Neuromethods Series Vol. 73 (Ed W Walz). Springer Science+Business Media, LLC, New York, NY, pp 385–403

    Google Scholar 

  92. Onimaru H, Kumagawa Y, Homma I (2006) Respiration-related rhythmic activity in the rostral medulla of newborn rats. J Neurophysiol 9:55–61

    Article  Google Scholar 

  93. Ruangkittisakul A, Panaitescu B, Kuribayashi J, Ballanyi K (2010) Caffeine reversal of ­opioid-evoked and endogenous inspiratory depression in perinatal rat en bloc medullas and slices. Adv Exp Med Biol 669:123–127

    Article  PubMed  CAS  Google Scholar 

  94. Bobocea N, Ruangkittisakul A, Ballanyi K (2010) Multiphoton/confocal Ca2+−imaging of inspiratory pre-Bötzinger complex neurons at the rostral or caudal surface of newborn rat brainstem slices. Adv Exp Med Biol 669:81–85

    Article  PubMed  CAS  Google Scholar 

  95. St. John WM (2009) Noeud vital for breathing in the brainstem: gasping-yes, eupnoea-doubtful. Philos Trans R Soc Lond B Biol Sci 364:2625–2633

    Article  PubMed  Google Scholar 

  96. Zanella S, Roux JC, Viemari JC, Hilaire G (2006) Possible modulation of the mouse respiratory rhythm generator by A1/C1 neurones. Respir Physiol Neurobiol 153:126–138

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Work contributing to this study was supported by the Alberta Heritage Foundation for Medical Research (AHFMR), Alberta Innovates Health Solutions-Hotchkiss Brain Institute (AIHS-HBI), the Canada Foundation for Innovation (CFI-ASRIP), and operating plus training grants from the Canadian Institutes of Health Research (CIHR).

Author information

Authors and Affiliations

  1. Faculty of Medicine & Dentistry, Department of Physiology and Centre for Neuroscience, University of Alberta, Edmonton, AB, Canada

    Araya Ruangkittisakul, Bogdan Panaitescu, Lucia Secchia, Nicoleta Bobocea, Chase Kantor, Junya Kuribayashi & Klaus Ballanyi

  2. Center for Medical Sciences, Ibaraki Prefectural University of Health Sciences, Ami, Ibaraki, Japan

    Makito Iizuka

Authors
  1. Araya Ruangkittisakul
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bogdan Panaitescu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lucia Secchia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nicoleta Bobocea
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chase Kantor
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Junya Kuribayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Makito Iizuka
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Klaus Ballanyi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Klaus Ballanyi .

Editor information

Editors and Affiliations

  1. Faculty of Medicine & Dentistry, Department of Physiology and Centre for, University of Alberta, N/A 232D, Edmonton, T6G 2S2, Alberta, Canada

    Klaus Ballanyi

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media New York

About this protocol

Cite this protocol

Ruangkittisakul, A. et al. (2012). Anatomically “Calibrated” Isolated Respiratory Networks from Newborn Rodents. In: Ballanyi, K. (eds) Isolated Central Nervous System Circuits. Neuromethods, vol 73. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-020-5_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-020-5_2

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-019-9

  • Online ISBN: 978-1-62703-020-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation