TY - JOUR
T1 - Correlation of actin crosslinker and capper expression levels with stereocilia growth phases
AU - Avenarius, Matthew R.
AU - Saylor, Katherine W.
AU - Lundeberg, Megan R.
AU - Wilmarth, Phillip A.
AU - Shin, Jung-Bum
AU - Spinelli, Kateri J.
AU - Pagana, James M.
AU - Andrade, Leonardo
AU - Kachar, Bechara
AU - Choi, Dongseok
AU - David, Larry L.
AU - Barr-Gillespie, Peter G.
N1 - Mass Spectrometry of Developing Cochlear Epithelia To determine the expression profile of actin crosslinkers and other actin-cytoskeleton proteins during development of the chick cochlea, we examined organs from E14 and 16, which represent the beginning and end of the diameter-widening phase (phase II), and 18 and E21, during and at the end of the renewed elongation phase (phase III).
PY - 2014/2
Y1 - 2014/2
N2 - During development of the chick cochlea, actin crosslinkers and barbed-end cappers presumably influence growth and remodeling of the actin paracrystal of hair cell stereocilia. We used mass spectrometry to identify and quantify major actin-associated proteins of the cochlear sensory epithelium from E14 to E21, when stereocilia widen and lengthen. Tight actin crosslinkers (i.e. fascins, plastins, and espin) are expressed dynamically during cochlear epithelium development between E7 and E21, with FSCN2 replacing FSCN1 and plastins remaining low in abundance. Capping protein, a barbed-end actin capper, is located at stereocilia tips; it is abundant during growth phase II, when stereocilia have ceased elongating and are increasing in diameter. Capping protein levels then decline during growth phase III, when stereocilia reinitiate barbed-end elongation. Although actin crosslinkers are readily detected by electron microscopy in developing chick cochlea stereocilia, quantitative mass spectrometry of stereocilia isolated from E21 chick cochlea indicated that tight crosslinkers are present there in stoichiometric ratios relative to actin that are much lower than their ratios for vestibular stereocilia. These results demonstrate the value of quantitation of global protein expression in chick cochlea during stereocilia development.
AB - During development of the chick cochlea, actin crosslinkers and barbed-end cappers presumably influence growth and remodeling of the actin paracrystal of hair cell stereocilia. We used mass spectrometry to identify and quantify major actin-associated proteins of the cochlear sensory epithelium from E14 to E21, when stereocilia widen and lengthen. Tight actin crosslinkers (i.e. fascins, plastins, and espin) are expressed dynamically during cochlear epithelium development between E7 and E21, with FSCN2 replacing FSCN1 and plastins remaining low in abundance. Capping protein, a barbed-end actin capper, is located at stereocilia tips; it is abundant during growth phase II, when stereocilia have ceased elongating and are increasing in diameter. Capping protein levels then decline during growth phase III, when stereocilia reinitiate barbed-end elongation. Although actin crosslinkers are readily detected by electron microscopy in developing chick cochlea stereocilia, quantitative mass spectrometry of stereocilia isolated from E21 chick cochlea indicated that tight crosslinkers are present there in stoichiometric ratios relative to actin that are much lower than their ratios for vestibular stereocilia. These results demonstrate the value of quantitation of global protein expression in chick cochlea during stereocilia development.
UR - https://www.scopus.com/pages/publications/84893217541
U2 - 10.1074/mcp.M113.033704
DO - 10.1074/mcp.M113.033704
M3 - Article
SN - 1535-9476
VL - 13
SP - 606
EP - 620
JO - Molecular and Cellular Proteomics
JF - Molecular and Cellular Proteomics
IS - 2
ER -