Topo Pneumonectomia Modello di crescita compensatoria Lung
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
La funzione principale del polmone è di fornire ossigeno e anidride carbonica scambio tra un organismo e l'atmosfera. Negli esseri umani, una serie di condizioni congenite ed acquisite portano a ridurre la superficie del polmone che si traduce in deficit della funzionalità respiratoria. Anche se una serie di terapie come corticosteroidi inalatori, broncodilatatori, ossigeno supplementare, e la ventilazione meccanica cronica sono utilizzati per attenuare le conseguenze di funzionalità polmonare ridotta 1-3, la terapia ideale per queste condizioni dovrebbe promuovere la ricrescita del tessuto polmonare funzionale – cioè, polmone rigenerazione.
Rigenerazione dei tessuti dei mammiferi è stata ben documentata. L'africano Mouse spinosi può rigenerare ampie aree di pelle senza formazione di cicatrici 4. La falange distale nell'uomo può rigenerarsi dopo una lesione o amputazione 5-7. Dopo pneumonectomia (PNX), la crescita del polmone di compensazione avviene nei topi 8, ratti 9, dogs 10, e gli esseri umani 11. Per definizione, la crescita del polmone di compensazione coinvolge non solo l'espansione di spazi aerei esistenti, ma ri-septation di questi spazi aerei allargata con l'espansione del microcircolo associato 12. Analisi di espressione genica ha dimostrato che questo modello ricapitola molti degli eventi di segnalazione dello sviluppo polmonare 13. Quattro settimane dopo il mouse PNX, superficie alveolare è equivalente a quella di fittizie animali operati 14. In questo manoscritto, descriviamo la PNX mouse e procedure PNX fittizi.
Protocol
Representative Results
Discussion
Abbiamo fornito la descrizione più dettagliata delle procedure PNX del mouse PNX e mouse farsa segnalati fino ad oggi. Abbiamo fatto il lettore a conoscenza di alcuni dei problemi comuni che gli investigatori di apprendimento la procedura comunemente incontrano, e abbiamo delineato diverse tecniche sviluppate dal nostro laboratorio per mitigare queste trappole. Altri laboratori che utilizzano questo modello possono sviluppare altre modifiche tecnica o utilizzare strumenti diversi. Nel valutare le differenze nelle tecni…
Divulgazioni
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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