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. 2017 Dec 12;4(6):ENEURO.0268-17.2017. doi: 10.1523/ENEURO.0268-17.2017

Figure 5.

Figure 5.

LTP-induced regulation of hnRNP K-associated mRNAs in vivo. Experiments were performed in live anesthetized rats. Electrodes were positioned for selective unilateral stimulation of the medial perforant path fibers in the angular bundle and recording of the evoked field potentials in the hilar region of the DG. A, Time course plots showing changes in the medial perforant path-evoked fEPSP slope expressed as percentage of baseline. Values are means ± SEM. Test pulses were applied at 0.033 Hz. The HFS paradigm (indicated by arrows) consisted of eight pulses of 400 Hz, repeated four times at 10-s intervals. Three sessions of HFS were given at intervals of five min. n = 6 for each time point. B, The variation of hnRNP K, GluN1, GluA1, and BDNF mRNA levels was assayed by qPCR using total RNA samples obtained from DG homogenates collected 30-min post-HFS and the nonstimulated contralateral control tissue. The results are presented as mean ± SEM normalized to the contralateral nonstimulated DG, and Hprt1 (hypoxanthine guanine phosphoribosyl transferase 1) was used as internal control gene. Results are the average ± SEM of six experiments (n = 6 DG) analyzed in three independent preparations; *p < 0.05 as determined by Student’s t test. C, The levels of hnRNP K, GluN1, GluA1, and BDNF transcripts coimmunoprecitated with hnRNP K were assayed by qPCR. hnRNP K protein was immunoprecipitated from equal amounts (500 μg) of total extracts from homogenized DG collected 30-min post-HFS and the contralateral tissue. Results are presented as mean ± SEM normalized to the contralateral DG and are the average ± SEM of six experiments (n = 6 DG) analyzed in three independent preparations; **p < 0.01; *p < 0.05 as determined by the Student’s t test. D, hnRNP K protein levels were measured by Western blotting using DG homogenate samples collected 30-min post-HFS and the contralateral nonstimulated tissue. The results are the average ± SEM of six experiments (n = 6 DG) analyzed in three independent preparations and are presented as the percentage change in hnRNP K protein levels in the treated DG relative to the nonstimulated contralateral tissue. GAPDH was used as loading control.