OBJECTIVE: The objectives of the current experiments were to develop and characterize primary rat nasal epithelial cultures and evaluate their usefulness as a model of cystic fibrosis (CF) sinonasal transepithelial transport and CF transmembrane conductance regulator (CFTR) function. STUDY DESIGN: Laboratory in vitro and animal studies. METHODS: CFTR+/+ and CFTR-/- rat nasal septal epithelia (RNSE) were cultured on semipermeable supports at an air-liquid interface to confluence and full differentiation. Monolayers were mounted in Ussing chambers for pharmacologic manipulation of ion transport and compared to similar filters containing murine (MNSE) and human (HSNE) epithelia. Histology and scanning electron microscopy (SEM) were completed. Real-time polymerase chain reaction of CFTR+/+ RNSE, MNSE, and HSNE was performed to evaluate relative CFTR gene expression. RESULTS: Forskolin-stimulated anion transport (ΔIsc in μA/cm2 ) was significantly greater in epithelia derived from CFTR+/+ when compared to CFTR-/- animals (100.9 ± 3.7 vs. 10.5 ± 0.9; P < 0.0001). Amiloride-sensitive ISC was equivalent (-42.3 ± 2.8 vs. -46.1 ± 2.3; P = 0.524). No inhibition of CFTR-mediated chloride (Cl- ) secretion was exhibited in CFTR-/- epithelia with the addition of the specific CFTR inhibitor, CFTRInh -172. However, calcium-activated Cl- secretion (UTP) was significantly increased in CFTR-/- RNSE (CFTR-/- -106.8 ± 1.6 vs. CFTR+/+ -32.2 ± 3.1; P < 0.0001). All responses were larger in RNSE when compared to CFTR+/+ and CFTR-/- (or F508del/F508del) murine and human cells (P < 0.0001). Scanning electron microscopy demonstrated 80% to 90% ciliation in all RNSE cultures. There was no evidence of infection in CFTR-/- rats at 4 months. CFTR expression was similar among species. CONCLUSION: The successful development of the CFTR-/- rat enables improved evaluation of CF sinus disease based on characteristic abnormalities of ion transport. LEVEL OF EVIDENCE: NA. Laryngoscope, 127:E384-E391, 2017.
OBJECTIVE: The objectives of the current experiments were to develop and characterize primary rat nasal epithelial cultures and evaluate their usefulness as a model of cystic fibrosis (CF) sinonasal transepithelial transport and CF transmembrane conductance regulator (CFTR) function. STUDY DESIGN: Laboratory in vitro and animal studies. METHODS:CFTR+/+ and CFTR-/- ratnasal septal epithelia (RNSE) were cultured on semipermeable supports at an air-liquid interface to confluence and full differentiation. Monolayers were mounted in Ussing chambers for pharmacologic manipulation of ion transport and compared to similar filters containing murine (MNSE) and human (HSNE) epithelia. Histology and scanning electron microscopy (SEM) were completed. Real-time polymerase chain reaction of CFTR+/+ RNSE, MNSE, and HSNE was performed to evaluate relative CFTR gene expression. RESULTS:Forskolin-stimulated anion transport (ΔIsc in μA/cm2 ) was significantly greater in epithelia derived from CFTR+/+ when compared to CFTR-/- animals (100.9 ± 3.7 vs. 10.5 ± 0.9; P < 0.0001). Amiloride-sensitive ISC was equivalent (-42.3 ± 2.8 vs. -46.1 ± 2.3; P = 0.524). No inhibition of CFTR-mediated chloride (Cl- ) secretion was exhibited in CFTR-/- epithelia with the addition of the specific CFTR inhibitor, CFTRInh -172. However, calcium-activated Cl- secretion (UTP) was significantly increased in CFTR-/- RNSE (CFTR-/- -106.8 ± 1.6 vs. CFTR+/+ -32.2 ± 3.1; P < 0.0001). All responses were larger in RNSE when compared to CFTR+/+ and CFTR-/- (or F508del/F508del) murine and human cells (P < 0.0001). Scanning electron microscopy demonstrated 80% to 90% ciliation in all RNSE cultures. There was no evidence of infection in CFTR-/- rats at 4 months. CFTR expression was similar among species. CONCLUSION: The successful development of the CFTR-/- rat enables improved evaluation of CF sinus disease based on characteristic abnormalities of ion transport. LEVEL OF EVIDENCE: NA. Laryngoscope, 127:E384-E391, 2017.
Authors: J R Riordan; J M Rommens; B Kerem; N Alon; R Rozmahel; Z Grzelczak; J Zielenski; S Lok; N Plavsic; J L Chou Journal: Science Date: 1989-09-08 Impact factor: 47.728
Authors: Scott D Grosse; Coleen A Boyle; Jeffrey R Botkin; Anne Marie Comeau; Martin Kharrazi; Margaret Rosenfeld; Benjamin S Wilfond Journal: MMWR Recomm Rep Date: 2004-10-15
Authors: Marcelo B Antunes; Bradford A Woodworth; Geeta Bhargave; Guoxiang Xiong; Jorge L Aguilar; Adam J Ratner; James L Kreindler; Ronald C Rubenstein; Noam A Cohen Journal: Biotechniques Date: 2007-08 Impact factor: 1.993
Authors: Katherine L Tuggle; Susan E Birket; Xiaoxia Cui; Jeong Hong; Joe Warren; Lara Reid; Andre Chambers; Diana Ji; Kevin Gamber; Kengyeh K Chu; Guillermo Tearney; Li Ping Tang; James A Fortenberry; Ming Du; Joan M Cadillac; David M Bedwell; Steven M Rowe; Eric J Sorscher; Michelle V Fanucchi Journal: PLoS One Date: 2014-03-07 Impact factor: 3.240
Authors: Shaoyan Zhang; Daniel Skinner; Stephen Bradley Hicks; Mark O Bevensee; Eric J Sorscher; Ahmed Lazrak; Sadis Matalon; Carmel M McNicholas; Bradford A Woodworth Journal: PLoS One Date: 2014-08-12 Impact factor: 3.240
Authors: Jacelyn E Peabody; Ren-Jay Shei; Brent M Bermingham; Scott E Phillips; Brett Turner; Steven M Rowe; George M Solomon Journal: Am J Physiol Lung Cell Mol Physiol Date: 2018-03-01 Impact factor: 5.464
Authors: Jessica Grayson; Kiranya E Tipirneni; Daniel F Skinner; Matthew Fort; Do-Yeon Cho; Shaoyan Zhang; Andrew C Prince; Dong-Jin Lim; Calvin Mackey; Bradford A Woodworth Journal: Int Forum Allergy Rhinol Date: 2017-06-08 Impact factor: 3.858
Authors: Kiranya E Tipirneni; Jessica W Grayson; Shaoyan Zhang; Do-Yeon Cho; Daniel F Skinner; Dong-Jin Lim; Calvin Mackey; Guillermo J Tearney; Steven M Rowe; Bradford A Woodworth Journal: Int Forum Allergy Rhinol Date: 2017-06-28 Impact factor: 3.858
Authors: Do-Yeon Cho; Daniel Skinner; Shaoyan Zhang; Ahmed Lazrak; Dong Jin Lim; Christopher G Weeks; Catherine G Banks; Chang Kyun Han; Si-Kwan Kim; Guillermo J Tearney; Sadis Matalon; Steven M Rowe; Bradford A Woodworth Journal: J Ginseng Res Date: 2019-09-13 Impact factor: 6.060