To study the expression of plasma membrane Ca(2+)-ATPase isoform 2 (PMCA2) and its alternative splicing at sites A (the first intracellular loop) and C (the C-terminal region) in the neonatal rat cochlea.
The cochleae from rats postnatal day 3 to postnatal day 4 (P3-P4) were dissected, fixed, embedded, and sectioned. Meanwhile, the cochlear coils from neonatal rats were isolated and fixed. Using immunofluorescence staining, the expression of PMCA2 was respectively examined in the cochlear sections and cochlear coils. In addition, the total RNAs of basilar membrane (BM, including the organ of corti, the same below), spiral ganglion (SG), spiral ligament (SL, including SV, the same below), and the whole cochlea from neonatal rats were respectively extracted and reverse transcribed to cDNAs, then subjected to primers flanking site A or C in the PMCA2 gene using reverse transcription polymerase chain reaction (RT-PCR). Western blot was also applied to detect the expression of PMCA2 isoforms in the cochlear tissues.
We found that PMCA2 is strongly expressed in outer hair cell (OHC) bundles, SG, and stria vascularis (SV), weakly expressed in Reissner's membrane (RM), and occasionally expressed in inner hair cell (IHC) bundles. Moreover, w/a is the major splice form of PMCA2 present in hair cell bundles, z/b and z/c are the major splice forms of PMCA2 present in SG, and w/a and w/c are the major splice forms of PMCA2 present in SV. In the whole cochlea, variants w, y, and z were detected at site A, and variants a, b, and c were detected at site C. Using Western blot, variant a or b was also detectable in the same cochlear tissues mentioned above.
PMCA2 and its splice variants at sites A and C are differentially expressed in cochlear tissues of neonatal rat.
International journal of pediatric otorhinolaryngology 02/2011; 75(2):196-201. DOI:10.1016/j.ijporl.2010.10.033 · 1.32 Impact Factor
Asthma is a common polygenic disease, caused by complex interactions between multiple genes and environmental factors. Study of the gene-gene interactions would contribute to a new insight into the pathogenesis and therapeutics of asthma.
To evaluate the single and combined associations of eight single-nucleotide polymorphisms loci in five candidate genes with the development of asthma in Chinese children.
We examined eight single-nucleotide polymorphisms (SNPs) in five key asthma susceptibility genes and performed single SNP association study, haplotype analysis, and gene-gene interactions analysis in 479 Chinese children, including 252 asthmatic subjects and 227 healthy controls. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Haplotype analysis was detected by SHEsis software. Gene-gene interactions were tested using the multifactor dimensionality reduction (MDR) method.
There were significant differences of interleukin (IL)-13 R130Q and IL-13 C1923T in genotype and allele frequency distributions between the asthmatic group and control group. Furthermore, the A allele of IL-13 R130Q and the T allele of IL-13 C1923T were significantly associated with increased risk of asthma (odds ratio [OR] = 1.59, 95% confidence interval [CI] 1.20-2.09, p = .0010; OR = 1.57, 95% CI 1.19-2.08, p = .0014, respectively). By haplotype analysis, the C-G and T-A haplotypes consisting of IL-13 C1923T and IL-13 R130Q and the G-A and A-A haplotypes consisting of IL-4Ralpha I75V and IL-4Ralpha Q576R were significantly associated with asthma (p < .05). Using MDR, the authors detected significant gene-gene interactions with a best six-locus model among IL-4 -C33T, IL-13 R130Q, IL-4Ralpha I75V, IL-4Ralpha Q576R, STAT6 C2892T, and CD14 -C159T on the risk of asthma (OR = 4.43, 95% CI 1.30-15.04, p < .001, by 1000-fold permutation test).
These data suggest that genetic variants in the IL-13 gene may play an important role in the development of pediatric asthma in Middle China. In addition, the significant gene-gene interactions among IL-4 -C33T, IL-13 R130Q, IL-4Ralpha I75V, IL-4Ralpha Q576R, STAT6 C2892T, and CD14 -C159T may increase an individual's susceptibility to asthma and contribute to the pathogenesis of asthma.
Journal of Asthma 04/2010; 47(3):238-44. DOI:10.3109/02770900903509099 · 1.83 Impact Factor
To investigate culturing neural stem cells (NSCs) from rat embryos in vitro and to observe their growth and differentiation.
NSCs were isolated from hippocampus of SD rat embryos (P16-P18) and cultured in DMEM/F12 medium containing EGF, bFGF, B27. To observe process of cell proliferation by microscope and identify cell types by immunocytochemical analyses after differentiation.
NSCs grew well in serum-free conditional medium and their cell bodies present transparent with good refraction at about eighth day. After differentiation, the cells demonstrated NSE and GFAP immunoreactive.
NSCs were cultured well in serum-free conditional medium and they could be induced to differentiate into neurons and astrocytes in serum conditional medium.
Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 09/2008; 22(16):747-50.