Optical Recording of Spontaneous Respiratory Neuron Activity in the Rat Brain Stem.

Tokumasu, Masayuki; Nakazono, Yoshimi; Ide, Hideto; Akagawa, Kimio; Onimaru, Hiroshi

doi:10.2170/jjphysiol.51.613

Cited by 15 publications

(15 citation statements)

References 23 publications

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“…For representation of the time course of fluorescence changes, averaged fluorescence intensity was assessed and inverted in a 50-pixel region of interest covering an area of 92 × 108µm. As discussed before, optical signals presumably reflect primarily the depolarization associated with respiratory drive potentials [35,61].…”

Section: Voltage-sensitive Dye-imagingmentioning

confidence: 82%

Opioids prolong and anoxia shortens delay between onset of preinspiratory (pFRG) and inspiratory (preBötC) network bursting in newborn rat brainstems

Ballanyi

Ruangkittisakul

Onimaru

2009

Pflugers Arch - Eur J Physiol

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Differential responses to opioids established the hypothesis that pre/postinspiratory (Pre-I) neurons of the parafacial respiratory group (pFRG) and inspiratory (Insp) neurons of the pre-Bötzinger complex (preBötC) constitute a dual brainstem respiratory center. For further analysis of pFRG/preBötC interactions, we studied in newborn rat brainstem-spinal cord preparations opioid and anoxia effects on histologically identified pFRG-driven "type-I" Insp preBötC neurons and Pre-I neurons from three distinct respiratory brainstem regions. The micro-opioid [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin (DAMGO) slowed inspiratory-related cervical nerve bursts quantally, whereas anoxia induced nonquantal slowing and repetitive cervical bursts. DAMGO had no effect on membrane potential or input resistance of Pre-I neurons, while anoxia hyperpolarized them (approximately 5 mV) and decreased their resistance (approximately 30%). DAMGO prolonged the preinspiratory phase of Pre-I neuron bursting, whereas anoxia caused a shift to postinspiratory (48%) or inspiratory (22%) activity and silenced further 30% of cells. Pre-I neuron responses were not correlated with their rostrocaudal location or morphology. Neither DAMGO nor anoxia changed membrane potential of type-I neurons, but decreased their input resistance by 33% and 21%, respectively. The opposite DAMGO- and anoxia-evoked phase shifts of Pre-I neuron activity were reflected by corresponding shifts of pre/postinspiratory drive potentials in type-I neurons and, partly, by voltage-sensitive dye-imaged medullary neuronal population activities. The findings suggest that opioids presynaptically delay activation of type-I neurons as the target of drive from the pFRG to the preBötC. Contrary, anoxia seems to partly synchronize the pFRG and preBötC rhythm generators. This may enhance inspiratory and postinspiratory medullary activities for triggering multiple inspiratory motor bursts.

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Section: Voltage-sensitive Dye-imagingmentioning

confidence: 82%

Opioids prolong and anoxia shortens delay between onset of preinspiratory (pFRG) and inspiratory (preBötC) network bursting in newborn rat brainstems

Ballanyi

Ruangkittisakul

Onimaru

2009

Pflugers Arch - Eur J Physiol

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“…2003; Tokumasu et al, 2001; Janczewski et al, 2002), has not been well characterized with respect to exact neurotransmitter phenotype, greatly limiting comparison between studies or conducting specific studies with an identified cell type. Although Onimaru and colleagues have evidence for both inhibitory (Arata et al, 1998) and excitatory-type neurons (Janczewski et al, 2002; Onimaru et al, 2006) in this area, they have provided only correlative physiological data rather than direct recording and histochemical identification of pFRG neurons.…”

Section: Brainstem Respiratory Areas With Identified Major Ionotromentioning

confidence: 99%

Identification of neurotransmitters and co-localization of transmitters in brainstem respiratory neurons

Stornetta

2008

Respiratory Physiology & Neurobiology

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Identifying the major ionotropic neurotransmitter in a respiratory neuron is of critical importance in determining how the neuron fits into the respiratory system, whether in producing or modifying respiratory drive and rhythm. There are now several groups of respiratory neurons whose major neurotransmitters have been identified and in some of these cases, more than one transmitter have been identified in particular neurons. This review will describe the physiologically identified neurons in major respiratory areas that have been phenotyped for major ionotropic transmitters as well as those where more than one transmitter has been identified. Although the purpose of the additional transmitter has not been elucidated for any of the respiratory neurons, some examples from other systems will be discussed.

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“…A recent study provides evidence that pacemaker activity is exclusively associated with gasping patterns in situ (27). The current hypotheses regarding respiratory rhythm generation are largely based on electrophysiological studies, which yield excellent temporal resolution of single-cell activities, but analysis of population activity is difficult.With the recent advances in imaging techniques, the PBC, as well as other respiratory centers, have been studied optically using voltage-sensitive dyes in isolated brain stem-spinal cord preparations (8,23,25,39) and even in the perfused brainstem preparation (9, 29). However, respiratory network functions and the activity of several network elements could not be probed at single-cell resolution.…”

mentioning

confidence: 99%

“…With the recent advances in imaging techniques, the PBC, as well as other respiratory centers, have been studied optically using voltage-sensitive dyes in isolated brain stem-spinal cord preparations (8,23,25,39) and even in the perfused brainstem preparation (9, 29). However, respiratory network functions and the activity of several network elements could not be probed at single-cell resolution.…”

mentioning

confidence: 99%

Imaging of respiratory-related population activity with single-cell resolution

Funke¹,

Dutschmann²,

Müller³

2007

American Journal of Physiology-Cell Physiology

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September 6, 2006; doi:10.1152/ajpcell.00253.2006.-The pre-Bötzinger complex (PBC) in the rostral ventrolateral medulla contains a kernel involved in respiratory rhythm generation. So far, its respiratory activity has been analyzed predominantly by electrophysiological approaches. Recent advances in fluorescence imaging now allow for the visualization of neuronal population activity in rhythmogenic networks. In the respiratory network, voltage-sensitive dyes have been used mainly, so far, but their low sensitivity prevents an analysis of activity patterns of single neurons during rhythmogenesis. We now have succeeded in using more sensitive Ca 2ϩ imaging to study respiratory neurons in rhythmically active brain stem slices of neonatal rats. For the visualization of neuronal activity, fluo-3 was suited best in terms of neuronal specificity, minimized background fluorescence, and response magnitude. The tissue penetration of fluo-3 was improved by hyperosmolar treatment (100 mM mannitol) during dye loading. Rhythmic population activity was imaged with single-cell resolution using a sensitive charge-coupled device camera and a ϫ20 objective, and it was correlated with extracellularly recorded mass activity of the contralateral PBC. Correlated optical neuronal activity was obvious online in 29% of slices. Rhythmic neurons located deeper became detectable during offline image processing. Based on their activity patterns, 74% of rhythmic neurons were classified as inspiratory and 26% as expiratory neurons. Our approach is well suited to visualize and correlate the activity of several single cells with respiratory network activity. We demonstrate that neuronal synchronization and possibly even network configurations can be analyzed in a noninvasive approach with single-cell resolution and at frame rates currently not reached by most scanning-based imaging techniques. neuronal network; fluorescence imaging; optical fingerprint; respiratory rhythm generation; glia A RHYTHMOGENIC NEURONAL NETWORK with intrinsic voltagedependent bursting properties is located in the pre-Bötzinger complex (PBC) of the rostral ventrolateral medulla (1,16,17,30,35). The respiratory rhythms of the in vitro PBC were previously studied in transverse slice preparations of 250-to 700-m thickness (30,32,35), tilted-sagittal slice preparations (28), as well as in en bloc preparations (22, 37). The location of the primary respiratory rhythm generator and a potential role for pacemaker neurons in the generation of breathing rhythms remain controversial though (13). A recent study provides evidence that pacemaker activity is exclusively associated with gasping patterns in situ (27). The current hypotheses regarding respiratory rhythm generation are largely based on electrophysiological studies, which yield excellent temporal resolution of single-cell activities, but analysis of population activity is difficult.With the recent advances in imaging techniques, the PBC, as well as other respiratory centers, have been studied optically using voltage-sensi...

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Optical Recording of Spontaneous Respiratory Neuron Activity in the Rat Brain Stem.

Cited by 15 publications

References 23 publications

Opioids prolong and anoxia shortens delay between onset of preinspiratory (pFRG) and inspiratory (preBötC) network bursting in newborn rat brainstems

Opioids prolong and anoxia shortens delay between onset of preinspiratory (pFRG) and inspiratory (preBötC) network bursting in newborn rat brainstems

Identification of neurotransmitters and co-localization of transmitters in brainstem respiratory neurons

Imaging of respiratory-related population activity with single-cell resolution

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