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Mouse Embryonic Lung Culture, A System to Evaluate the Molecular Mechanisms of Branching
Published on: June 30, 2010
Expression of growth hormone and its receptor in the lungs of embryonic chicks
Jason A Beyea1, David M Olson, Richard A Vandergriend
1Department of Physiology and Perinatal Research Centre, University of Alberta, Edmonton, T6G 2H7, Canada.
Insights
The embryonic lung produces growth hormone (GH) before pituitary secretion, suggesting local roles in lung development. This prenatal GH is distinct from pituitary GH and acts via GH receptors in the lung.
Area of Science:
- Endocrinology
- Developmental Biology
- Pulmonology
Background:
- The lung is a known postnatal target for growth hormone (GH), with dysfunction observed in GH excess or deficiency.
- However, lung development occurs before the pituitary gland's GH secretion, suggesting alternative GH sources or roles during this critical period.
Purpose of the Study:
- To investigate the possibility of local GH production within the embryonic lung.
- To determine the characteristics and localization of GH and its receptor in the developing embryonic chick lung.
Main Methods:
- Reverse transcription/polymerase chain reaction (RT-PCR) to detect GH mRNA in embryonic lung tissue.
- In situ hybridization to localize GH mRNA and GH immunoreactivity within lung cells.
- Immunohistochemistry to identify GH protein localization and characteristics.
Main Results:
- GH mRNA, similar to pituitary GH transcript, was found in embryonic chick lungs from days 11 to 18 of incubation.
- GH immunoreactivity was localized to mesenchymal and epithelial cells, and later to smooth muscle around blood vessels and bronchioles.
- Lung GH was associated with a 15-kDa protein, differing from the pituitary's 26-kDa form. GH receptor mRNA and immunoreactivity were also abundant.
Conclusions:
- The embryonic lung synthesizes GH locally, potentially acting in an autocrine or paracrine manner to influence lung development and function.
- This local GH production precedes pituitary GH secretion, highlighting its importance in prenatal lung maturation.
- The presence of GH receptors further supports a functional role for lung-derived GH during embryonic development.
Abstract:
The lung is well established as being a postnatal target site for growth hormone (GH) action, since pathophysiological states of GH excess and deficiency are both associated with impaired pulmonary function. Pituitary GH is therefore probably involved in normal lung growth or development, although perinatal lung development occurs prior to the differentiation of pituitary somatotrophs and the ontogeny of pituitary GH secretion. The lung itself may, however, be a site of GH production during prenatal development, since a specific GH-response gene (a marker of GH activity) is expressed in the lungs of early chick embryos, in which GH immunoreactivity is widespread in many other peripheral tissues. We have assessed this possibility in embryonic chicks. A 690-bp cDNA, identical in size and nucleotide sequence to the full-length pituitary GH transcript, was amplified by reverse transcription/polymerase chain reaction from total RNA extracted from the lungs of embryos at 11, 13, 15, and 18 days of the 21-day incubation period. This transcript was localized by in situ hybridization to mesenchymal and epithelial cells of the developing lungs, in which specific GH immunoreactivity was similarly located. Intense GH immunoreactivity was also present after embryonic day 15 (ED15) in the smooth muscle surrounding blood vessels in the lung and surrounding the bronchioles. Lung GH immunoreactivity was primarily associated with a 15-kDa protein, rather than the 26-kDa protein in the pituitary gland. After the onset of pituitary GH secretion (at ED17), GH mRNA was barely detectable in the lungs of ED20 embryos, at the start of lung breathing. GH immunoreactivity was, however, still present in some cells in the lungs of ED20 embryos. GH-receptor mRNA and immunoreactivity were also widespread and abundant within the embryonic lung. Lung GH may thus have autocrine or paracrine roles in lung development or in pulmonary function prior to the ontogeny of the pituitary gland and the appearance of GH in peripheral plasma.
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