We investigated the role of TrkB and the localization of BDNF in the adrenal gland during immobilization stress for 60 min

We investigated the role of TrkB and the localization of BDNF in the adrenal gland during immobilization stress for 60 min. feedback loop in the adrenal medulla during immobilization stress. that chromaffin cells of the adrenal medulla showed significantly increased TrkB mRNA and protein levels after 60 min of stress (Kondo hybridization (ISH) and immunohistochemistry (IHC). In experiments, we compared between intact rats and sham-operated rats in each group. Blood sampling After deep anesthesia of all rats at the end of experiments, blood samples were collected between 07:00 hours and 11:00 hours by cardiac puncture into Venoject II? tubes containing EDTA (Terumo, Tokyo, Japan). Tubes were immediately placed on ice, and centrifuged at 760 for 15 min at 4C. Plasma was stored at ?20C prior to HPLC (Saruta transcription of linearized pGEM-T Easy Vectors (Promega Co., Madison, WI, USA) (Ottem 0.05; = 4 per group; error bars = SD). (b) Significant differences in dopamine levels were seen between the agonist group and all other groups. Data were analyzed by the MannCWhitney 0.05; = 4 per group; error bars Docetaxel (Taxotere) = SD). Adrenaline was not detected in any group. catecholamine levels after stimulation using TrkB agonist antibody and 60-min stress We examined the release of catecholamine in rats subjected to 60 min of TrkB stimulation via TrkB agonist antibody (anti-TrkB; clone 47/TrkB). Adrenaline levels in the various groups were as follows: control group, 431.5 247.6 pg/mL; stress group, 3260.5 462.9 pg/mL; stress+agonist group, 17689.5 8676.9 pg/mL; and stress (adrenalectomy)+agonist group, 2.500 1.300 pg/mL. There were significant differences between the Docetaxel (Taxotere) adrenaline levels in each Docetaxel (Taxotere) group (Fig. 2a). Noradrenaline levels in the various groups were as follows: control group, 1357.0 677.7 pg/mL; stress group, 6053.8 1585.8 pg/mL; stress+agonist group, 13737.8 3754.9 pg/mL; and stress (adrenalectomy)+agonist group, 3550.8 614.7 pg/mL. Similarly, there were Docetaxel (Taxotere) significant differences between the noradrenaline levels in each group (Fig. 2b). Dopamine levels in the various groups were as follows: control group, 61.5 5.2 pg/mL; stress group, 254.0 62.5 pg/mL; stress+agonist group, 405.5 111.0 pg/mL; and stress (adrenalectomy)+agonist group, 338.5 78.5 pg/mL. There were significant differences in dopamine levels between the control group and those in all other groups (Fig. 2c). Open in a separate windows Fig. 2 catecholamine levels Lox after stimulation with tyrosine receptor kinase B (TrkB) expressed in the adrenal medulla after 60-min stress using TrkB agonist antibody. Rats were stimulated with TrkB agonist antibody after 60-min immobilization stress. Levels of adrenaline (a), noradrenaline (b) and dopamine (c) in blood were assayed by HPLC. (a) Significant differences in adrenaline levels were seen between each group. Data were analyzed by the MannCWhitney 0.05, ** 0.01; = 4C6 per group; error bars = SD). (b) Significant differences in noradrenaline levels were seen between each group. Data were analyzed by the MannCWhitney 0.05, ** 0.01; = 4C6 per group; error bars = SD). (c) Significant differences in dopamine levels were seen between the control group and all other groups, but there were Docetaxel (Taxotere) no significant differences between the stress+agonist and stress groups, or between the stress+agonist and stress (adrenalectomy)+agonist groups. Data were analyzed by the MannCWhitney 0.05, ** 0.01; = 4C6 per group; error bars = SD). In the control, stress and stress+agonist groups, there were no significant differences between intact rats and sham-operated rats. Furthermore, in the stress (adrenalectomy)+agonist group, there were no significant differences between sham-operated rats and stress+agonist group (data not shown). Expression of TrkB mRNA and BDNF mRNA in adrenal gland during 60-min stress We further analyzed TrkB mRNA and BDNF mRNA expression in the adrenal glands during 60-min immobilization stress using RT-PCR. After 60 min of stress, in the stress and stress+agonist groups, BDNF mRNA was markedly up-regulated in the adrenal gland when compared with the control group. Similarly, TrkB mRNA was weakly up-regulated when compared with the control group (Fig. 3). There were no differences in expression between intact rats and sham-operated rats (data not shown). Open.