Long-term treatment with rapamycin inhibited the growth of body weight and immune organs in young rats.

<p>Rats of 2 weeks of age were administrated different doses of rapamycin for 4 weeks. A. Body weight monitoring demonstrated that treatment with rapamycin at 1.0 and 3.0 mg/kg decreased body weight significantly as compared to control rats 3 weeks after treatment. B. Rats were sacrificed after 4 weeks of treatment and the ratio of spleen to body weight was calculated. Rats treated with 1.0 and 3.0 mg/kg exhibited the ratio decrease. C. Rats were sacrificed after 4 weeks of treatment and the ratio of thymus to body weight was calculated. A decreased ratio was also found in rats treated with 1.0 and 3.0 mg/kg rapamycin. *p<0.05; **p<0.01 compared to control group. n = 15 rats/ group.</p

The key proteins of mTOR pathway are developmentally regulated.

<p>Representative blots of Western blot analysis of Akt, mTOR, S6 and their phosophorylation forms during brain development in hippocampus (A) and in neocortex (B). C. Quantitative summary of individual proteins in hippocampus demonstrated that p-Akt, p-mTOR and p-S6 were low at postnatal 1 d and transiently increased from 3 d to 2 w (p-Akt) or to 3 w (p-mTOR, p-S6). D. Quantitative summary of individual proteins in neocortex demonstrated that p-Akt, p-mTOR and p-S6 remained at a high level for the first 2 postnatal (p-S6) or 3 weeks (p-Akt and p-mTOR) and decreased thereafter. * p<0.05 compared to postnatal 1 d. n = 6 rats/ group.</p

Long-term treatment with rapamycin results in decrease in cellular immune factors.

<p>Rats of 2 weeks of age were administrated different doses of rapamycin for 4 weeks and sacrificed. Cortical brain tissue (A, C, E, G) and blood (B, D, F, H) were collected and used for ELISA examination of Il-1β, IL-2, IFN-γ and TNF-α. No obvious change was noted among different groups in Il-1β both in brain and blood (A,B). IL-2 was decreased with 1.0 and 3.0 mg/kg rapamycin treatment both in brain and blood (C, D). IFN-γ was only decreased with 3.0 mg/kg rapamycin treatment both in brain and blood (E, F). All rapamycin-treated groups exhibited a decrease in TNF-α in brain (G), while the decrease was only shown in 3.0 mg/kg of rapamycin treatment in blood (H). *p<0.05; **p<0.01 compared to control group. n = 6 rats/ group.</p

Long-term treatment with rapamycin affects spatial learning.

<p>Rats of 2 weeks of age were administrated different doses of rapamycin for 4 weeks and subjected to Morris water maze experiment. A. Rats injected with 1.0 and 3.0 mg/kg rapamycin had prolonged escape latency during training days 3 and 4. Rapamycin-treated rats at the dose of 1.0 and 3.0 mg/kg resulted in significant increase in escape latency (B) and swimming length (C) and decrease in number of crossing the target (D) on the 5th day. *p<0.05 compared to control group. n = 15 rats/ group.</p

Long-term treatment with rapamycin results in cognitive impairment.

<p>Rats of 2 weeks of age were administrated different doses of rapamycin for 4 weeks and subjected to Y maze experiment after Morris water maze. A. The percentage of correct times during the training. It was only decreased in rats treated with 3.0 mg/kg rapamycin on the 3rd and 4th day. B. The percentage of correct times on the 5th day. C. The average latency to the safe area was markedly increased in 1.0 and 3.0 mg/kg rapamycin-treated rats. D. The percentage of active escape before electric shock was significantly decreased in 1.0 and 3.0 mg/kg rapamycin-treated rats. *p<0.05 compared to control group. n = 15 rats/ group.</p

Long-term treatment with rapamycin results in anxiety-like behavior.

<p>Rats of 2 weeks of age were administrated different doses of rapamycin for 4 weeks and subjected to open field experiment after Y maze. Rats treated with 1.0 and 3.0 mg/kg rapamycin demonstrated decreased number of crossings (A), increased fecal pellets (B), retention time in the central area (C) and freezing time (D). *p<0.05 compared to control group. n = 15 rats/ group.</p

Long-term treatment with rapamycin results in decrease in progenitor cells.

<p>A-L. Representative images of immunohistochemical staining of DAPI, DCX and their merge. M. Quantitative calculation of DCX-positive cells per field in different groups. Rats treated with 1.0 and 3.0 mg/kg rapamycin demonstrated decrease in DCX-positive cells. *p<0.05; **p<0.01 compared to control group. Scale bar = 100 um. n = 6 rats/ group.</p

Rapamycin inhibits S6 phosphorylation dose-dependently in young rats.

<p>Rats of 2 weeks of age were administrated rapamycin for 3 d (A, C) or for 4 w (B, D). Representative blots in neocortex of short- (A) or long- treatment (B) with rapamycin at different doses. C. Quantitative summary demonstrated that 3 days treatment of 0.03 mg/kg rapamycin had no effect on p-S6, whereas 0.1, 0.3 and 3.0 inhibited p-s6 dose-dependently. D. Quantitative summary of 4 weeks treatment demonstrated that only 1.0 and 3.0 mg/kg rapamycin inhibited p-S6. *p<0.05, **p<0.01, ***p<0.001 compared to control group. n = 6 rats/ group.</p

Alignment of the amino acid sequence of TaRab7 and selected Rab7 proteins.

<p>Alignment of the amino acid sequence of <i>Triticum aestivum</i> Rab7 (<i>TaRab7</i>) with other Rab7 proteins from <i>Oryza sativa</i> (<i>OsRab7</i>), <i>Hordeum vulgare</i> (<i>HvRab7</i>), <i>Arabidopsis</i> (<i>RabG3b</i>), <i>Homo sapiens</i> (<i>HsRab7</i>) and <i>Saccharomyces cerevisiae</i> (<i>ScYpt7</i>). Sequences were aligned using DNAMAN. Identical residues in all organisms are shaded. Red underlines indicate sequence motifs involved in nucleotide binding and hydrolysis that are conserved in Rab GTPases.</p

Histological observations during the incompatible interaction between wheat and the stripe rust fungus when the transcription of <i>TaRab7</i> was repressed.^a

a<p>Abbreviations: hpi.: hours post inoculation.</p>b<p>Distance from the base of substomatal vesicles to hyphal tips. Values with * are significantly different at P = 0.05 according to the Tukey's test.</p>c<p>BSMV: γ and BSMV: TaRab7, leaves inoculated with BSMV: γ or BSMV: TaRab7 followed by infection with CYR23.</p