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Meeting abstracts
Neural control of breathing
An Official Satellite of the International Congress of Physiological Sciences (IUPS) 2001,
Rotorua, New Zealand, 1–4 September 2001
Received: 2 August 2001
Published: 17 August 2001
Respir Res 2001, 2 (suppl 1):S1–S37
© 2001 BioMed Central Ltd
(Print ISSN 1465-9921; Online ISSN 1465-993X)
ORAL PRESENTATIONS — SESSION 1
Ontogeny and phylogeny of respiratory control
1.1
Early development of respiratory rhythm
generation in mice and chicks
J Champagnat , G Fortin, S Jungbluth, V Abadie, F Chatonnet,
E Dominquez-del-Toro, L Guimarães
UPR 2216 (Neurobiologie Génétique et Intégrative), IFR 2118 (Institut
de Neurobiologie Alfred Fessard), CNRS, 91198, Gif-sur-Yvette, France
Breathing in mammals starts in the foetus and acquires a vital
importance at birth. The ability to produce rhythmic motor behav-
iours linked to respiratory function is a property of the brainstem
reticular formation, which has been remarkably conserved during
the evolution of vertebrates. Therefore, to understand the biological
basis of the breathing behavior, we are investigating conservative
developmental mechanisms orchestrating the organogenesis of the
brainstem. In vertebrates, the hindbrain is one of the vesicles that
appears at the anterior end of the neural tube of the embryo.
Further morphogenetic subdivision ensues whereby the hindbrain
neuroepithelium becomes partitioned into an iterated series of
compartments called rhombomeres. The segmentation process is
believed to determine neuronal fates by encoding positional infor-
mation along the rostro-caudal axis. Before and at the onset of seg-
mentation, genes encoding transcription factors such as Hox,
Krox-20, kreisler, are expressed in domains corresponding to the
limits of future rhombomeres. Inactivation of these genes specifi-
cally disturbs the rhombomeric pattern of the hindbrain. The pre-
sentation will address the problem of whether this primordial
rhombomeric organisation influences later function of respiratory
control networks in chicks and mice.
Experiments were performed in embryos and after birth in trans-
genic mice. They show that, although expression of developmental
genes and hindbrain segmentation are transient events of early
embryonic development, they are important for the process of res-
piratory rhythm generation by brainstem neuronal networks. We
have found in chick that at the end of the period of segmentation,
the hindbrain contains neuronal rhythm generators that conform to
the rhombomeric anatomical pattern. We have also identified a
minimal rhombomeric motif allowing the post-segmental maturation
of a specific (GABAergic) rhythm-promoting circuit. Furthermore,
in vivo and in vitro analysis of neurons in transgenic mice revealed
postnatal respiratory phenotypes associated with defects of central
pontine and/or afferent respiratory control in Krox-20, Hoxa1 and
kreisler mutants. Neonatal respiratory phenotypes are also induced
in mice by treatment with low doses of retinoic acid that slightly
change the early embryonic development of the Pons. Altogether,
these experiments indicate that segmentation-related specifica-
tions of the hindbrain rhythmic neuronal network influences the res-
piratory patterns after birth. Therefore, early developmental
processes have to be taken into account to understand normal and
pathological diversity of the breathing behaviour in vertebrates.
Acknowledgement: Supported by HFSP RG101/97, ACI (BDPI)
2000, CEE BIO4CT, ICCTI PRAXIS XXI (BD/11299/97).
1.2
Development of gill and lung breathing in amphibia
MJ Gdovin , VV Jackson, JC Leiter
Division of Life Sciences, University of Texas at San Antonio, TX, USA
In the 25 morphological stages of larval bullfrog development there
exists a gradual transition from gill to lung ventilation associated
with a developmental decrease in the contribution of the skin in
gas exchange. Bath application of GABA and/or glycine inhibited
gill but not lung burst activities of cranial nerve (CN) VII in the
premetamorphic (stages 16–19) in vitro tadpole brainstem prepa-
ration [1]. It was proposed that the neural basis of gill rhythmogen-
esis involved network inhibition, whereas lung rhythmicity was
pacemaker driven [1]. Bath application of a bicuculline/strychnine
solution abolished gill and enhanced lung bursting in stages
16–19 in vitro [1]. Bath application of the GABA
B
receptor antago-
nist 2-hydroxy-saclofen disinhibited the lung central pattern genera-
tor (CPG) resulting in precocious lung bursting patterns as early as
developmental stage 6 [2].
We recorded efferent activity from CN VII and spinal nerve (SN) II in
the in vitro tadpole brainstem preparation in three successive devel-
opmental groups (3–9; 10–15; 16–19) before and after bath appli-
cation of a 10 μM bicuculline and 5 μM strychnine solution. We also
exposed the brainstem to bath pH 7.4, 7.8, and 8.2 before and after
bath application of bicuculline/strychnine. Bicuculline/strychnine
produced lung ventilatory bursts in all developmental stages tested,
indicating the presence of the lung CPG as well as excitatory
synapses to respiratory motor neurons as early as stage 3.
We also designed an experiment to examine the importance of
lung ventilation on the developmental shift from gill to lung burst-
ing. Two groups of tadpoles were hatched from eggs. Control tad-