PMID- 35604150
OWN - NLM
STAT- MEDLINE
DCOM- 20220525
LR  - 20240416
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 183
DP  - 2022 May 5
TI  - Utilizing the Precision-Cut Lung Slice to Study the Contractile Regulation of 
      Airway and Intrapulmonary Arterial Smooth Muscle.
LID - 10.3791/63932 [doi]
AB  - Smooth muscle cells (SMC) mediate the contraction of the airway and the 
      intrapulmonary artery to modify airflow resistance and pulmonary circulation, 
      respectively, hence playing a critical role in the homeostasis of the pulmonary 
      system. Deregulation of SMC contractility contributes to several pulmonary 
      diseases, including asthma and pulmonary hypertension. However, due to limited 
      tissue access and a lack of culture systems to maintain in vivo SMC phenotypes, 
      molecular mechanisms underlying the deregulated SMC contractility in these 
      diseases remain fully identified. The precision-cut lung slice (PCLS) offers an 
      ex vivo model that circumvents these technical difficulties. As a live, thin lung 
      tissue section, the PCLS retains SMC in natural surroundings and allows in situ 
      tracking of SMC contraction and intracellular Ca(2+) signaling that regulates SMC 
      contractility. Here, a detailed mouse PCLS preparation protocol is provided, 
      which preserves intact airways and intrapulmonary arteries. This protocol 
      involves two essential steps before subjecting the lung lobe to slicing: 
      inflating the airway with low-melting-point agarose through the trachea and 
      infilling pulmonary vessels with gelatin through the right ventricle. The PCLS 
      prepared using this protocol can be used for bioassays to evaluate 
      Ca(2+)-mediated contractile regulation of SMC in both the airway and the 
      intrapulmonary arterial compartments. When applied to mouse models of respiratory 
      diseases, this protocol enables the functional investigation of SMC, thereby 
      providing insight into the underlying mechanism of SMC contractility deregulation 
      in diseases.
FAU - Bai, Yan
AU  - Bai Y
AD  - Division of Newborn Medicine, Department of Pediatrics, Massachusetts General 
      Hospital and Harvard Medical School; ybai4@bwh.harvard.edu.
FAU - Ai, Xingbin
AU  - Ai X
AD  - Division of Newborn Medicine, Department of Pediatrics, Massachusetts General 
      Hospital and Harvard Medical School.
LA  - eng
GR  - R01 HL132991/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Video-Audio Media
DEP - 20220505
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Arteries
MH  - Lung
MH  - Mice
MH  - *Muscle Contraction/physiology
MH  - *Muscle, Smooth/physiology
MH  - Myocytes, Smooth Muscle/physiology
EDAT- 2022/05/24 06:00
MHDA- 2022/05/26 06:00
CRDT- 2022/05/23 09:05
PHST- 2022/05/23 09:05 [entrez]
PHST- 2022/05/24 06:00 [pubmed]
PHST- 2022/05/26 06:00 [medline]
AID - 10.3791/63932 [doi]
PST - epublish
SO  - J Vis Exp. 2022 May 5;(183). doi: 10.3791/63932.