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Intermittent Fasting Dietary Restriction Regimen Negatively Influences Reproduction in Young Rats: A Study of Hypothalamo-Hypophysial-Gonadal Axis

Figure 7

Representative immunofluorescent staining, Western blot and RT-PCR data of GnRH and PSA-NCAM in male rats.

Confocal immunofluorescent images of 30 µ thick coronal sections from median eminence-arcuate (ME) region of ad libitum fed (AL) and Intermittent fasting-dietary restricted (IF-DR) male (n = 5 each) rat brain. The immunostaining in the ME region for GnRH and PSA-NCAM is shown for GnRH (A and B), PSA-NCAM (C and D) as well as dual immunofluorescence of GnRH and PSA-NCAM (E and F) for AL and IF-DR animals, respectively. Both GnRH (A and B) and PSA-NCAM (C and D) immunostaining was reduced in IF-DR animals as compared to their corresponding control rats. (K) Western blot hybridization for PSA-NCAM and α-tubulin from median eminence region of AL and IF-DR male rats. (M) RT-PCR results for PST and actin from median eminence region of AL and IF-DR male rats. (I) depicts relative intensity measurement ±SEM of GnRH immunofluorescence, performed by an observer blind to the experiments. (J) depict relative intensity measurement ±SEM of PSA-NCAM immunofluorescence from ME region. (L and N) depict relative optical density ±SEM of PSA-MCAM western blot analysis and PST mRNA expression from median eminence region. *pValue<0.05. Scale bar = 60 µ (A–F).

Figure 7

doi: https://doi.org/10.1371/journal.pone.0052416.g007