ࡱ> 02/q` -bjbjqPqP 4::- 2222$V nnnnnIII2444444$h X II Xnnmz z z nn2z 2z z z nb d[0#28 "z 20z Z z z IvTz DW/IIIXXd III d22 BDNF Expression in Lymphoblastoid Cell Lines Carrying BDNF SNPs Associated with Bipolar Disorder YongLin Gao1, MD, Mathew Galante1, James El-Mallakh1, Zhenmin Lei2, MD, Rif S. El-Mallakh1, MD Departments of Psychiatry 1 Departments of Obstetrics and Gynecology2 University of Louisville School of Medicine, Louisville, Kentucky Background: Alterations in brain derived neurotrophic factor (BDNF) expression have been associated with mood disturbances. Specific single nucleotide polymorphisms (SNPs) of the BDNF gene have been reported to be associated with the psychiatric condition of bipolar disorder. However, the functional consequences of these SNPs are unknown. Methods: Lymphoblastoid cell lines (n = 30) obtained from bipolar type I subjects who participated in a study examining genetic linkage in bipolar illness were utilized. Cells expressing variations in a total of 10 SNPs, five of which are linked with bipolar disorder, were exposed to the pro-apoptotic stressors of serum deprivation (to eliminate exogenous BDNF) alone, or serum deprivation combined with the sodium ionophore, monensin (1 M), for 24 hrs, to induce BDNF production. Intracellular BDNF precursor (pro-BDNF) and mature BDNF (mBDNF) were quantified with immunoblot analysis. Sodium concentration was measured with flame spectrophotometry. Apoptosis was confirmed with DNA fragmentation analysis. Results: Serum deprivation alone for 24 hrs did not induce apoptosis, but combination of serum deprivation and monensin was associated with apoptosis within 24 hrs. Apoptosis was associated with an elevation of intracellular sodium concentrations to 1.65 times above baseline. Monensin treatment with serum deprivation is associated with an increase of mBDNF compared to serum deprivation alone (2.110.05 vs 1.980.03,P<0.05), but proBDNF levels were not different. There were no differences in either pro-BDNF or mBDNF related to any of the 10 SNPs that have been reported to associate with a risk of bipolar illness. Conclusions: In current study, neither intracellular proBDNF, nor mBDNF protein levels varied with the 10 BDNF SNPs that have been reported to be associated with a risk of bipolar disorder. 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