Mus Pneumonectomy Model af kompenserende Lung Vækst
Summary
Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.
Abstract
In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.
Introduction
Den vigtigste funktion af lungen er at sørge for ilt og kuldioxid udveksling mellem en organisme og atmosfæren. Hos mennesker en masse af medfødte og erhvervede betingelser føre til nedsat lunge overfladeareal, hvilket resulterer i forringet lungefunktion. Selv om en række behandlinger såsom inhalerede kortikosteroider, bronkodilatatorer, supplerende ilt og kronisk mekanisk ventilation anvendes til at afbøde følgerne af nedsat lungefunktion 1-3, vil den ideelle behandling for disse tilstande fremmer genvækst af funktionel lungevæv – dvs. lunge regenerering.
Pattedyrvæv regenerering er veldokumenteret. Den afrikanske Spiny Mouse kan regenerere store områder af huden uden ardannelse 4. Den distale phalanx i mennesker kan regenerere efter skade eller amputation 5-7. Efter pneumonectomy (PNX) forekommer kompenserende lunge vækst i mus 8 rotter 9, gørgs 10, og mennesker 11. Per definition kompenserende lunge vækst indebærer ikke kun udvidelse af eksisterende luftrum, men re-septumdannelse af disse udvidede luftrum med udvidelse af den tilhørende mikrocirkulationen 12. Genekspressionsanalyse har vist, at denne model sammenfatter mange af de signaler begivenheder lunge udvikling 13. Fire uger efter mus PNX, alveolære overflade svarer til den for skinopererede dyr 14. I dette manuskript, beskriver vi musen PNX og fingeret PNX procedurer.
Protocol
Representative Results
Discussion
Vi har givet den mest detaljerede beskrivelse af de mus PNX og mus fingeret PNX procedurer rapporteret til dato. Vi har gjort læseren opmærksom på flere af de almindelige faldgruber at efterforskerne lære proceduren sædvanligvis støder, og vi har skitseret en række teknikker udviklet af vores laboratorium for at afbøde disse faldgruber. Andre laboratorier anvender denne model kan have udviklet andre teknik modifikationer eller bruge forskellige instrumenter. Ved evalueringen forskelle i teknikker, vil de enkelte…
Disclosures
The authors have nothing to disclose.
Acknowledgements
The authors would like to acknowledge the Cincinnati Children’s Hospital Division of Veterinary Services for their assistance. This manuscript was supported by the National Institutes of Health K12 HD028827. Anna Perl PhD taught the authors this surgical procedure.
Materials
| Name of the reagent | Company | Catalogue number | Comments (optional) |
| 6-inch Vascular Clip Applicator | Teleflex Medical (WECK) | 137062 | |
| Horizon Small Titanium Red Clip | Teleflex Medical (WECK) | 1201 | |
| Narrow Pattern 12cm Curved Forceps | Fine Science Tools | 11003-12 | |
| Curved Serated 10 cm Graefe Forceps | Fine Science Tools | 11052-10 | |
| Castroviejo Needle Holder | Fine Science Tools | 12565-14 | |
| Straight 9 cm Strabismus Scissors (Blunt Tip) | Fine Science Tools | 14075-09 | |
| Straight 8.5 cm Hardened Fine Scissors | Fine Science Tools | 14090-09 | |
| Straight, Blunt Tip Cohan-Vannas Spring Scissors | Fine Science Tools | 15000-12 | |
| Skin Glue | Gluture | 32046 | |
| 22 ga angiocatheter | |||
| 24 ga angiocatheter | |||
| 3 mL luer lock syringe | |||
| 4 short retractors | |||
| 2 long retractors | |||
| 5-0 Prolene on curved cutting needle | Ethicon | 8698G | |
| 0.5 mL syringe on 27 ga needle | |||
| Normal Saline | |||
| Buprenorphine | |||
| Press-n-Seal Wrap | Glad Products Company | ||
| 12 inch X 12 inch cork board stack | Office Depot | ||
| 70% Ethanol | |||
| Betadine | |||
| Mouse Ventilator | Hugo Sachs Elektronnik | Minivent Type 845 | |
| Isoflurane Vaporizer | OHMEDA | Excel 210 SE | |
| artificial tear ointment | puralube | NDC: 17033-211-38 |
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