Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension
<p>Abstract</p> <p>Background</p> <p>Chronic hypoxia influences gene expression in the lung resulting in pulmonary hypertension and vascular remodelling. For specific investigation of the vascular compartment, laser-microdissection of intrapulmonary arteries was combine...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2005-09-01
|
| Series: | Respiratory Research |
| Online Access: | http://respiratory-research.com/content/6/1/109 |
| _version_ | 1818140806020595712 |
|---|---|
| author | Ziegler Andreas Koenig Inke R Laumanns Isabel Wolff Stephanie Wilhelm Jochen Kwapiszewska Grazyna Seeger Werner Bohle Rainer M Weissmann Norbert Fink Ludger |
| author_facet | Ziegler Andreas Koenig Inke R Laumanns Isabel Wolff Stephanie Wilhelm Jochen Kwapiszewska Grazyna Seeger Werner Bohle Rainer M Weissmann Norbert Fink Ludger |
| author_sort | Ziegler Andreas |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Chronic hypoxia influences gene expression in the lung resulting in pulmonary hypertension and vascular remodelling. For specific investigation of the vascular compartment, laser-microdissection of intrapulmonary arteries was combined with array profiling.</p> <p>Methods and Results</p> <p>Analysis was performed on mice subjected to 1, 7 and 21 days of hypoxia (FiO<sub>2 </sub>= 0.1) using nylon filters (1176 spots). Changes in the expression of 29, 38, and 42 genes were observed at day 1, 7, and 21, respectively. Genes were grouped into 5 different classes based on their time course of response. Gene regulation obtained by array analysis was confirmed by real-time PCR. Additionally, the expression of the growth mediators PDGF-B, TGF-β, TSP-1, SRF, FGF-2, TIE-2 receptor, and VEGF-R1 were determined by real-time PCR. At day 1, transcription modulators and ion-related proteins were predominantly regulated. However, at day 7 and 21 differential expression of matrix producing and degrading genes was observed, indicating ongoing structural alterations. Among the 21 genes upregulated at day 1, 15 genes were identified carrying potential hypoxia response elements (HREs) for hypoxia-induced transcription factors. Three differentially expressed genes (S100A4, CD36 and FKBP1a) were examined by immunohistochemistry confirming the regulation on protein level. While FKBP1a was restricted to the vessel adventitia, S100A4 and CD36 were localised in the vascular tunica media.</p> <p>Conclusion</p> <p>Laser-microdissection and array profiling has revealed several new genes involved in lung vascular remodelling in response to hypoxia. Immunohistochemistry confirmed regulation of three proteins and specified their localisation in vascular smooth muscle cells and fibroblasts indicating involvement of different cells types in the remodelling process. The approach allows deeper insight into hypoxic regulatory pathways specifically in the vascular compartment of this complex organ.</p> |
| first_indexed | 2024-12-11T10:49:50Z |
| format | Article |
| id | doaj.art-478ee5bd813449e6a8649c6edcd23635 |
| institution | Directory Open Access Journal |
| issn | 1465-9921 |
| language | English |
| last_indexed | 2024-12-11T10:49:50Z |
| publishDate | 2005-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj.art-478ee5bd813449e6a8649c6edcd236352022-12-22T01:10:21ZengBMCRespiratory Research1465-99212005-09-016110910.1186/1465-9921-6-109Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertensionZiegler AndreasKoenig Inke RLaumanns IsabelWolff StephanieWilhelm JochenKwapiszewska GrazynaSeeger WernerBohle Rainer MWeissmann NorbertFink Ludger<p>Abstract</p> <p>Background</p> <p>Chronic hypoxia influences gene expression in the lung resulting in pulmonary hypertension and vascular remodelling. For specific investigation of the vascular compartment, laser-microdissection of intrapulmonary arteries was combined with array profiling.</p> <p>Methods and Results</p> <p>Analysis was performed on mice subjected to 1, 7 and 21 days of hypoxia (FiO<sub>2 </sub>= 0.1) using nylon filters (1176 spots). Changes in the expression of 29, 38, and 42 genes were observed at day 1, 7, and 21, respectively. Genes were grouped into 5 different classes based on their time course of response. Gene regulation obtained by array analysis was confirmed by real-time PCR. Additionally, the expression of the growth mediators PDGF-B, TGF-β, TSP-1, SRF, FGF-2, TIE-2 receptor, and VEGF-R1 were determined by real-time PCR. At day 1, transcription modulators and ion-related proteins were predominantly regulated. However, at day 7 and 21 differential expression of matrix producing and degrading genes was observed, indicating ongoing structural alterations. Among the 21 genes upregulated at day 1, 15 genes were identified carrying potential hypoxia response elements (HREs) for hypoxia-induced transcription factors. Three differentially expressed genes (S100A4, CD36 and FKBP1a) were examined by immunohistochemistry confirming the regulation on protein level. While FKBP1a was restricted to the vessel adventitia, S100A4 and CD36 were localised in the vascular tunica media.</p> <p>Conclusion</p> <p>Laser-microdissection and array profiling has revealed several new genes involved in lung vascular remodelling in response to hypoxia. Immunohistochemistry confirmed regulation of three proteins and specified their localisation in vascular smooth muscle cells and fibroblasts indicating involvement of different cells types in the remodelling process. The approach allows deeper insight into hypoxic regulatory pathways specifically in the vascular compartment of this complex organ.</p>http://respiratory-research.com/content/6/1/109 |
| spellingShingle | Ziegler Andreas Koenig Inke R Laumanns Isabel Wolff Stephanie Wilhelm Jochen Kwapiszewska Grazyna Seeger Werner Bohle Rainer M Weissmann Norbert Fink Ludger Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension Respiratory Research |
| title | Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension |
| title_full | Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension |
| title_fullStr | Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension |
| title_full_unstemmed | Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension |
| title_short | Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension |
| title_sort | expression profiling of laser microdissected intrapulmonary arteries in hypoxia induced pulmonary hypertension |
| url | http://respiratory-research.com/content/6/1/109 |
| work_keys_str_mv | AT zieglerandreas expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT koeniginker expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT laumannsisabel expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT wolffstephanie expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT wilhelmjochen expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT kwapiszewskagrazyna expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT seegerwerner expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT bohlerainerm expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT weissmannnorbert expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension AT finkludger expressionprofilingoflasermicrodissectedintrapulmonaryarteriesinhypoxiainducedpulmonaryhypertension |