[Expression of plasma membrane Ca²(+)-ATPase isoform 2 in spiral ganglion cells of newborn rats].
OBJECTIVE
To study the expression of plasma membrane Ca(2+)-ATPase isoform 2 (PMCA2) in spiral ganglion cell (SGC) from inner ear of newborn rats and further check PMCA2 splice variants at site A and C.


METHODS
Spiral ganglion tissues isolated from cochlea of newborn rats (P3-P4) were cultured and identified in vitro. The cochlea of newborn rats (P3-P4) were isolated and cut into frozen sections. The expression of PMCA2 was detected by immunofluorescence analyses. The SGC cultured in 4 wells of the 6-well culture plate were collected and the total RNA was extracted by Trizol and reverse transcribed to cDNA. The site A and C splice variants of PMCA2 were respectively checked by nested PCR and common PCR.


RESULTS
The SGC grew well with good refraction and showed positive immunoreactivity for neuronal marker NF-200. Strong green fluorescence could be seen in cytomembrane, cytoplasm and neuritis, as well showing SGC immunoreactivity for PMCA2 antibody. In the cochlear sections, the spiral ganglion tissues were strongly stained by PMCA2. PMCA2z was present at splice site A, but PMCA2b and PMCA2c were present at splice site C.


CONCLUSIONS
SGC from newborn rats strongly expresses PMCA2 and different splice variants are present at PMCA2 splice site A and C.
Chen
Qing-guo
QG
Department of Otorhinolaryngology Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Chu
Han-qi
HQ
Wang
Xian-hong
XH
Chen
Jin
J
Li
Jian-ling
JL
Cui
Yong-hua
YH
D000818:Animals; D005260:Female; D008297:Male; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D022821:RNA Splice Sites; D051381:Rats; D017207:Rats, Sprague-Dawley; D013136:Spiral Ganglion / Q000166:cytology / Q000378:metabolism*
D020033:Protein Isoforms; D022821:RNA Splice Sites; D053499:Plasma Membrane Calcium-Transporting ATPases