Mus Pneumonektomi Modell av Kompenserende Lung Vekst
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 prinsipielle funksjon av lungen, er å sørge for oksygen og karbondioksyd utveksling mellom en organisme og atmosfæren. Hos mennesker, en rekke medfødte og ervervede tilstander føre til redusert lungeflateareal som resulterer i nedsatt lungefunksjon. Selv om en rekke behandlinger som inhalasjonssteroider, bronkodilatorer, ekstra oksygen, og kronisk mekanisk ventilasjon brukes for å dempe konsekvensene av svekket lungefunksjon 1-3, ville det ideelle behandling for disse forholdene fremme gjenvekst av funksjonell lungevev – dvs. lunge regenerering.
Pattedyrvev regenerering har vært godt dokumentert. The African Spiny Mouse kan regenerere store områder av huden uten arrdannelse 4. Den distale falanks hos mennesker kan regenerere etter skade eller amputasjon 5-7. Etter pneumonectomy (PNX), oppstår kompenserende lunge vekst i mus 8, rotter 9, gjøregs 10, og mennesker 11. Per definisjon innebærer kompenserende lunge vekst ikke bare utvidelse av eksisterende luftrommene, men re-tverrdeling av disse forstørrede luftrom med ekspansjon av den tilhørende mikrosirkulasjonen 12. Genekspresjonsanalyser har vist at denne modellen sammenfatter mange av de signal hendelsene i lunge utvikling 13. Fire uker etter mus PNX, er alveolar areal som tilsvarer humbug operert dyr 14. I dette manuskriptet, beskriver vi muse PNX og humbug PNX prosedyrer.
Protocol
Representative Results
Discussion
Vi har gitt den mest detaljerte beskrivelsen av mus PNX og mus humbug PNX prosedyrer rapportert til dags dato. Vi har gjort leseren oppmerksom på flere av de vanligste fallgruvene som etterforskerne læring prosedyren ofte støter på, og vi har skissert flere teknikker utviklet av vårt laboratorium for å redusere mot disse fallgruvene. Andre laboratorier som benytter denne modellen kan ha utviklet andre teknikk modifikasjoner eller bruke ulike instrumenter. Når man skal vurdere forskjeller i teknikker, vil 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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