Abstract
We recently developed an ex vivo lung slice model that allows for confocal live cell imaging (LCI) of neuroepithelial bodies (NEBs) in postnatal mouse lungs (postnatal days 1–21 and adult). NEBs are morphologically well-characterized, extensively innervated groups of neuroendocrine cells in the airway epithelium, which are shielded from the airway lumen by ‘Clara-like’ cells. The prominent presence of differentiated NEBs from early embryonic development onwards, strongly suggests that NEBs may exert important functions during late fetal and neonatal life. The main goal of the present study was to adapt the current postnatal LCI lung slice model to enable functional studies of fetal mouse lungs (gestational days 17–20).
In vibratome lung slices of prenatal mice, NEBs could be unequivocally identified with the fluorescent stryryl pyridinium dye 4-Di-2-ASP. Changes in the intracellular free calcium concentration and in mitochondrial membrane potential could be monitored using appropriate functional fluorescent indicators (e.g. Fluo-4).
It is clear that the described fetal mouse lung slice model is suited for LCI studies of Clara cells, ciliated cells, and the NEB microenvironment, and offers excellent possibilities to further unravel the significance of NEBs during the prenatal and perinatal period.
This work was supported by the following research grants:
A fellowship from the Agency for Innovation by Science and Technology in Flanders (IWT) to Robrecht Lembrechts (SB 81162), by grants of the Fund for Scientific Research-Flanders (FWO; G.0081.08 to D.A. and I.B., G.0589.11 to D.A. and J-P.T.), and by grants of the University of Antwerp (GOA BOF 2007 to D.A. and KP BOF 2011 to I.B.).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Adriaensen D, Brouns I, Van Genechten J, Timmermans J-P (2003) Functional morphology of pulmonary neuroepithelial bodies: extremely complex airway receptors. Anat Rec 270A:25–40
Adriaensen D, Brouns I, Pintelon I, De Proost I, Timmermans J-P (2006) Evidence for a role of neuroepithelial bodies as complex airway sensors: comparison with smooth muscle-associated airway receptors. J Appl Physiol 101:960–970
Bollé T, Lauweryns JM, Van Lommel A (2000) Postnatal maturation of neuroepithelial bodies and carotid body innervation: a quantitative investigation in the rabbit. J Neurocytol 29:241–248
Brouns I, Van Genechten J, Burnstock G, Timmermans J-P, Adriaensen D (2003) Ontogenesis of P2X3 receptor-expressing nerve fibres in the rat lung, with special reference to neuroepithelial bodies. Biomed Res 14:80–86
Cutz E, Jackson A (1999) Neuroepithelial bodies as airway oxygen sensors. Respir Physiol 115:201–214
De Proost I, Pintelon I, Brouns I, Kroese ABA, Riccardi D, Kemp PJ, Timmermans J-P, Adriaensen D (2008) Functional live cell imaging of the pulmonary neuroepithelial body microenvironment. Am J Respir Cell Mol Biol 39:180–189
De Proost I, Pintelon I, Wilkinson WJ, Goethals S, Brouns I, Van Nassauw L, Riccardi D, Timmermans J-P, Kemp PJ, Adriaensen D (2009) Purinergic signaling in the pulmonary neuroepithelial body microenvironment unraveled by live cell imaging. FASEB J 23:1153–1160
Fu XW, Wang D, Nurse CA, Dinauer MC, Cutz E (2000) NADPH oxidase is an O2 sensor in airway chemoreceptors: evidence from K+ current modulation in wild-type and oxidase-deficient mice. Proc Natl Acad Sci U S A 97:4374–4379
Fukuda J, Ishimine H, Masaki Y (2003) Long-term staining of live Merkel cells with FM dyes. Cell Tissue Res 311:325–332
Gosney JR (1993) Pulmonary neuroendocrine cells in species at high altitude. Anat Rec 236:105–107
Hoyt RF, McNelly NA, Sorokin SP (1993) Calcitonin gene-related peptide (CGRP) as regional mitogen for tracheobronchial epithelium of organ cultured fetal rat lungs. Am Rev Resp Dis 147:A498
Knowles MR, Clarke LL, Boucher RC (1991) Activation by extracellular nucleotides of chloride secretion in the airway epithelia of patients with cystic fibrosis. N Engl J Med 325:533–538
Liberati TA, Randle MR, Toth LA (2010) In vitro lung slices: a powerful approach for assessment of lung pathophysiology. Expert Rev Mol Diagn 10:501–508
Linnoila RI (2006) Functional facets of the pulmonary neuroendocrine system. Lab Invest 86:425–444
Pan J, Yeger H, Cutz E (2004) Innervation of pulmonary neuroendocrine cells and neuroepithelial bodies in developing rabbit lung. J Histochem Cytochem 52:379–389
Pintelon I, De Proost I, Brouns I, Van Herck H, Van Genechten J, Van Meir F, Timmermans J-P, Adriaensen D (2005) Selective visualisation of neuroepithelial bodies in vibratome slices of living lung by 4-Di-2-ASP in various animal species. Cell Tissue Res 321:21–33
Sanderson MJ (2011) Exploring lung physiology in health and disease with lung slices. Pulm Pharmacol Ther 24:452–465
Sorokin SP, Hoyt RF (1990) On the supposed function of neuroepithelial bodies in adult mammalian lungs. News Physiol Sci 5:89–95
Sorokin SP, Hoyt RF, Shaffer MJ (1997) Ontogeny of neuroepithelial bodies: correlations with mitogenesis and innervation. Microsc Res Tech 37:43–61
Van Lommel A, Bolle T, Fannes W, Lauweryns JM (1999) The pulmonary neuroendocrine system: the past decade. Arch Histol Cytol 62:1–16
Widdicombe JG (2001) Airway receptors. Respir Physiol 125:3–15
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media Dordrecht
About this paper
Cite this paper
Schnorbusch, K., Lembrechts, R., Brouns, I., Pintelon, I., Timmermans, JP., Adriaensen, D. (2012). Precision-Cut Vibratome Slices Allow Functional Live Cell Imaging of the Pulmonary Neuroepithelial Body Microenvironment in Fetal Mice. In: Nurse, C., Gonzalez, C., Peers, C., Prabhakar, N. (eds) Arterial Chemoreception. Advances in Experimental Medicine and Biology, vol 758. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-4584-1_22
Download citation
DOI: https://doi.org/10.1007/978-94-007-4584-1_22
Published:
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-007-4583-4
Online ISBN: 978-94-007-4584-1
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)