concanavalin-a and luzindole

A total of 360 post-hatching day 0 (P0) Arbor Acre male broilers, including intact, sham operation and pinealectomy groups, were exposed to white light (WL), red light (RL), green light (GL) and blue light (BL) from a light-emitting diode (LED) system until for P14. We studied the effects of melatonin and its receptors on monochromatic light-induced T-lymphocyte proliferation in the thymus of broilers. The density of proliferating cell nuclear antigen (PCNA) cells and the proliferation of T-lymphocytes in response to Concanavalin A (ConA) in GL significantly increased both in vivo and in vitro (from 9.57% to 32.03% and from 34.30% to 50.53%, respectively) compared with other lights (p<0.005) and was strongly correlated with melatonin levels in plasma (p<0.005). Pinealectomy reduced the levels of circulatory melatonin and the proliferation of T-lymphocytes and eliminated the differences between GL and other lights (p<0.005). However, exogenous melatonin (10(-9)M) significantly increased the proliferative activity of T-lymphocyte by 9.64% (p=0.002). In addition, GL significantly increased mRNA expression levels of Mel1a, Mel1b and Mel1c receptors from 21.09% to 32.57%, and protein expression levels from 24.43% to 42.92% compared with RL (p<0.05). However, these effects were blocked after pinealectomy. Furthermore, 4P-PDOT (a selective Mel1b antagonist) and prazosin (a selective Mel1c antagonist) attenuated GL-induced T-lymphocyte proliferation in response to ConA (p=0.000). Luzindole (a nonselective Mel1a/Mel1b antagonist), however, did not induce these effects (p=0.334). These results suggest that melatonin may mediate GL-induced T-lymphocyte proliferation via the Mel1b and Mel1c receptors but not via the Mel1a receptor.

Topics: Animals; Cell Proliferation; Cells, Cultured; Chickens; Concanavalin A; Gene Expression; Immunohistochemistry; Light; Male; Melatonin; Prazosin; Proliferating Cell Nuclear Antigen; Receptors, Melatonin; RNA, Messenger; T-Lymphocytes; Tetrahydronaphthalenes; Thymus Gland; Tryptamines

Despite the evidence for melatonin membrane receptors (MT1R and MT2R) on lymphoid tissues in a wide range of seasonal breeders, their specific potency has never been compared and correlated with cell-mediated immunity.. We used luzindole, a nonselective MT2R antagonist, and 4-phenyl-2-propionamidotetralin (4P-PDOT), a selective MT2R antagonist, to assess the potency of the melatonin receptors MT1 and MT2 in melatonin-induced immunity under both in vivo as well as in vitro conditions.. Physiological doses (25 μg/100 g body weight in vivo and 100 and 500 pg/ml in vitro) of melatonin upregulated both MT1R and MT2R expression as well as splenocyte proliferation, while higher doses (100 and 500 μg/100 g body weight in vivo and 1 ng/ml in vitro) downregulated splenocyte proliferation and the expression of both receptors. Luzindole antagonized the expression of both MT1R and MT2R in a dose-dependent manner under in vivo as well as in vitro conditions, while 4P-PDOT blocked the expression of MT2R only during both experimental conditions. Splenocyte proliferation and IL-2 secretion (in vitro) followed the MT1R expression pattern, while the MT2R expression pattern showed no definite relation with either splenocyte proliferation or IL-2 secretion under in vivo and in vitro conditions.. Immune function in tropical rodents is directly regulated by melatonin via its high-affinity membrane receptor MT1. MT1R plays a directive role in mediating splenocyte proliferation and IL-2 release, while the MT2R subtype appears not to be required for the immunoenhancing role of melatonin.

Topics: Analysis of Variance; Animals; Breeding; Cell Proliferation; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Gene Expression Regulation; Immunity; In Vitro Techniques; Interleukin-2; Lymphocytes; Male; Melatonin; Mitogens; Random Allocation; Receptor, Melatonin, MT1; Sciuridae; Seasons; Spleen; Tetrahydronaphthalenes; Tryptamines

In addition to marked seasonal changes in reproductive, metabolic, and other physiological functions, many vertebrate species undergo seasonal changes in immune function. Despite growing evidence that photoperiod mediates seasonal changes in immune function, little is known regarding the neuroendocrine mechanisms underlying these changes. Increased immunity in short days is hypothesized to be due to the increase in the duration of nightly melatonin secretion, and recent studies indicate that melatonin acts directly on immune cells to enhance immune parameters. The present study examined the contribution of melatonin receptors in mediating the enhancement of splenocyte proliferation in response to the T cell mitogen Concanavalin A in mice. The administration of luzindole, a high-affinity melatonin receptor antagonist, either in vitro or in vivo significantly attenuated the ability of in vitro melatonin to enhance splenic lymphocyte proliferation during the day or night. In the absence of melatonin or luzindole, splenocyte proliferation was intrinsically higher during the night than during the day. In the absence of melatonin administration, luzindole reduced the ability of spleen cells to proliferate during the night, when endogenous melatonin concentrations are naturally high. This effect was not observed during the day, when melatonin concentrations are low. Taken together, these results suggest that melatonin enhancement of splenocyte proliferation is mediated directly by melatonin receptors on splenocytes and that there is diurnal variation in splenocyte proliferation in mice that is also mediated by splenic melatonin receptors.

Topics: Animals; Cell Division; Cells, Cultured; Circadian Rhythm; Concanavalin A; Darkness; Light; Lymphocyte Activation; Male; Melatonin; Mice; Mice, Inbred C57BL; Photoperiod; Receptors, Cell Surface; Receptors, Cytoplasmic and Nuclear; Receptors, Melatonin; Spleen; T-Lymphocytes; Tryptamines