•Green light enhanced cClock, cBmal1 and cCry1 expression in chick hypothalamus.•Green light enhanced CLOCK and BMAL1 expression in chick hypothalamus.•CLOCK was located in the SCN, AM, PHN, PVN and ME in chick hypothalamus.To clarify the effect of monochromatic light on circadian clock gene expression in chick hypothalamus, a total 240 newly hatched chickens were reared under blue light (BL), green light (GL), red light (RL) and white light (WL), respectively. On the post-hatched day 14, 24-h profiles of seven core clock genes (cClock, cBmal1, cBmal2, cCry1, cCry2, cPer2 and cPer3) were measured at six time points (CT 0, CT 4, CT 8, CT 12, CT 16, CT 20, circadian time). We found all these clock genes expressed with a significant rhythmicity in different light wavelength groups. Meanwhile, cClock and cBmal1 showed a high level under GL, and followed a corresponding high expression of cCry1. However, RL decreased the expression levels of these genes. Be consistent with the mRNA level, CLOCK and BMAL1 proteins also showed a high level under GL. The CLOCK-like immunoreactive neurons were observed not only in the SCN, but also in the non-SCN brain region such as the nucleus anterior medialis hypothalami, the periventricularis nucleus, the paraventricular nucleus and the median eminence. All these results are consistent with the auto-regulatory circadian feedback loop, and indicate that GL may play an important role on the circadian time generation and development in the chick hypothalamus. Our results also suggest that the circadian clock in the chick hypothalamus such as non-SCN brain region were involved in the regulation of photo information.

•Mel1a were distributed in the POA, POP and SCN, Mel1b were distributed in the POA and SCN.•Green light was increased the expressions of Mel1a and Mel1b.•Melatonin played a critical role in green light-enhanced expressions of Mel1a and Mel1b.To study the mechanism of the effect of monochromatic light on physiological function in chicken, a total of 192 newly hatched chicks were randomly divided into intact, sham-operated and pinealectomy groups then exposed to white light (WL), red light (RL), green light (GL) and blue light (BL) using a light-emitting diode (LED) system for two weeks. At P14, the hypothalami were immediately collected for immunohistochemical staining of melatonin receptor subtypes (Mel1a and Mel1b) and detection of Mel1a and Mel1b expressions using RT-PCR and western blot. Immunohistochemical staining of the hypothalamus showed that the Mel1a-ir cells were distributed in the preoptic area (POA), nucleus preopticus periventricularis (POP) and suprachiasmatic nuclei (SCN), and the Mel1b-ir cells were presented in the POA and SCN. Analysis of RT-PCR and western blot showed that the mRNA and protein levels of Mel1a and Mel1b in the hypothalamus of chick exposed to GL were increased by 10.7–29.3%, 9.18–35.9% and 8.97–27.3% compared to those in the chicks exposed to WL (P = 0.029–0.002), RL (P = 0.027–0.001) and BL (P = 0.038–0.007) in the intact group, respectively. After pinealectomy, however, these parameters decreased and there were no significant differences among the WL, RL, GL and BL groups. These findings suggested that melatonin plays a critical role in GL illumination-enhanced Mel1a and Mel1b expressions in the hypothalamus of chicks.

Previous study has demonstrated that melatonin plays a critical role in monochromatic-light-induced lymphocyte proliferation in response to T cell mitogen concanavalin A (ConA). However, its intracellular mechanism is still unclear. In this study, we investigate the intracellular signal pathways of melatonin receptor-mediated T-lymphocyte proliferation in the spleens of chicks exposed to different light wavelengths. Results showed that green light enhanced T-lymphocyte proliferation by 2.46–6.83% and increased splenic mRNA and protein expressions of melatonin receptor subtypes (Mel1a, Mel1b and Mel1c) by 16.05–40.43% compared with the white, red and blue light groups. However, pinealectomy resulted in a decrease in T-lymphocyte proliferation and melatonin receptor expression with no statistically significant differences between the different light groups. In vitro experiments showed that the Mel1b selective antagonist 4P–PDOT, the Mel1c selective antagonist prazosin and the mitogen-activated protein kinase kinase-1 (MEK-1) inhibitor PD98059 suppressed both melatonin-induced lymphocyte proliferation in response to ConA and melatonin- and ConA-stimulated extracellular signal-regulated kinase 1/2 (ERK1/2) activity but that the Mel1a/Mel1b non-selective antagonist luzindole did not. In addition, pretreatment with forskolin (FSK, the adenylyl cyclase activator), H89 (the PKA inhibitor), U73122 (the PLC inhibitor) or Go6983 (the broad spectrum PKC inhibitor) markedly attenuated melatonin- and ConA-stimulated T-lymphocyte proliferation and ERK1/2 activity. These results demonstrate that melatonin mediates green-light-induced T-lymphocyte proliferation via the Mel1b and Mel1c receptors by triggering crosstalk between the cAMP/PKA and PLC/PKC signal pathways followed by ERK1/2 activation.

•Green light enhanced positive clock gene expression and decreased negative ones.•Red light increased negative clock genes expression and decreased positive ones.•Green light lifted melatonin level, but red light suppressed that in plasma.•Green and red light affected melatonin level by modulating clock gene expression.•Blue light advanced acrophases of negative clock genes and delayed positive ones.The avian pineal gland is a master clock that can receive external photic cues and translate them into output rhythms. To clarify whether a shift in light wavelength can influence the circadian expression in chick pineal gland, a total of 240 Arbor Acre male broilers were exposed to white light (WL), red light (RL), green light (GL) or blue light (BL). After 2 weeks light illumination, circadian expressions of seven core clock genes in pineal gland and the level of melatonin in plasma were examined. The results showed after illumination with monochromatic light, 24 h profiles of all clock gene mRNAs retained circadian oscillation, except that RL tended to disrupt the rhythm of cCry2. Compared to WL, BL advanced the acrophases of the negative elements (cCry1, cCry2, cPer2 and cPer3) by 0.1–1.5 h and delayed those of positive elements (cClock, cBmal1 and cBmal2) by 0.2–0.8 h. And, RL advanced all clock genes except cClock and cPer2 by 0.3–2.1 h, while GL delayed all clock genes by 0.5–1.5 h except cBmal2. Meanwhile, GL increased the amplitude and mesor of positive and reduced both parameters of negative clock genes, but RL showed the opposite pattern. Although the acrophase of plasma melatonin was advanced by both GL and RL, the melatonin level was significantly increased in GL and decreased in RL. This tendency was consistent with the variations in the positive clock gene mRNA levels under monochromatic light and contrasted with those of negative clock genes. Therefore, we speculate that GL may enhance positive clock genes expression, leading to melatonin synthesis, whereas RL may enhance negative genes expression, suppressing melatonin synthesis.

•Green light illumination better enhances T-lymphocyte proliferation in chick thymus.•Melatonin plays a critical role in green light-induced T-lymphocyte proliferation.•Mel1b and Mel1c receptors involve in green light-induced T-lymphocyte proliferation.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.

•The monochromatic green light enhanced B lymphocyte proliferation in young broilers.•Melatonin plays a critical role on monochromatic GL enhanced B cell proliferation.•The antioxidative pathway involved in MEL mediating GL induced B cell proliferation.To study the role of melatonin in the monochromatic light-induced B lymphocyte proliferation of bursa, a total of 360 newly hatched broilers, including intact, sham-operated, and pinealectomized groups, were exposed to blue light (BL), green light (GL), red light (RL) and white light (WL) from a light-emitting diode system for 14d. Both in vivo and in vitro studies showed that the percentage of proliferating cell nuclear antigen (PCNA)-immunoreactive lymphocytes and the lymphocyte proliferation in response to lipopolysaccharide in the bursa of broilers in the GL intact group was the highest values among the different intact groups with altered plasma melatonin levels. Additionally, the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and total antioxidant capability (T-AOC) were the highest, whereas malondialdehyde (MDA) content and inducible nitric oxide synthase (iNOS) expression were significantly decreased in the GL intact group. After pinealectomy, the levels of SOD, GSH-Px and T-AOC decreased remarkably in the various light-treatment groups, whereas the MDA content and iNOS expression significantly increased. The administration of exogenous melatonin (250 pg/mL) in vitro also significantly enhanced the bursal B lymphocyte proliferation. These findings suggest that GL illumination effectively elevates the antioxidative capacity to promote B lymphocyte proliferation of bursa in young broilers, which might depend on enhanced melatonin secretion.

•We examine changes in the antioxidative capacity and apoptosis-related proteins•Reduced antioxidative capacity involved in infertility of mice•Quinestrol induced testicular spermatogenic apoptosisThe effects of quinestrol on spermatogenesis were investigated in adult male mice by daily intragastric administration of quinestrol with various doses of 5, 10, 50 and 100 mg/kg body weight for 10 days. The sperm counts declined while the number of abnormal spermatozoa went up in mice treated with quinestrol. The testicular weight and seminiferous tubular area gradually declined with increasing dosages of quinestrol to 50 and 100 mg/kg. Rarefied germ cells showed irregular distributions in the seminiferous tubules of mice treated with 50 and 100 mg/kg quinestrol. Apoptosis was highly pronounced in spermatogonia, spermatocytes, spermatids and Leydig cells. Antioxidant enzyme activities – superoxide dismutase and glutathione peroxidase – as well as total antioxidant capacity significantly reduced, while malondialdehyde contents increased. The number of germ cells expressing caspase-3, p53, Bax and FasL significantly increased whereas cells expressing Bcl-2 significantly decreased in groups treated with 50 and 100 mg/kg quinestrol compared with the control. The concentration of nitrogen monoxidum also increased significantly under these dosages. The results suggest that quinestrol stimulates oxidative stress to induce apoptosis in spermatogenic cells through the mitochondrial and death receptor pathways in adult male mice.

This study determined the effects of melatonin (MEL) and its receptors on monochromatic light-induced bursal B-lymphocyte proliferation in broiler chickens. In vivo, green light (GL) enhanced the proliferation of B lymphocytes in bursas by 16.49% to 30.83% and the expression of MEL receptor subtypes 1a (Mel1a), Mel1b, and Mel1c receptors in bursas by 6.91% to 366.98% than other light colors. However, pinealectomy reduced these parameters and eliminated the differences between GL and other light groups. In vitro, the MEL-induced bursal B-lymphocyte proliferation was most suppressed by prazosin (P = 0.001, selective Mel1c antagonist), followed by luzindole (P = 0.022, nonselective Mel1a/Mel1b antagonist), but not by 4-phenyl-2-propionamideotetralin (P = 0.144, selective Mel1b antagonist). Similarly, dibutyryl-cyclic adenosine monophosphate (cAMP; analog of cAMP; P = 0.017) but not 8-(4-chloro-phenylthio)-2′-O-methyladenosine-3′,5′-cyclic monophosphate (P = 0.736; activator of exchange protein directly activated by cAMP) significantly inhibited bursal B-lymphocyte proliferation. These results suggest that MEL mediates GL-induced bursal B-lymphocyte proliferation through Mel1c and Mel1a receptors but not Mel1b receptors by activating the cAMP/protein kinase A pathway.