Photic Regulation of Circadian Rhythms and Voluntary Ethanol Intake: Role of Melanopsin-expressing Intrinsically Photosensitive Retinal Ganglion Cells

“Non-image-forming” (NIF) effects of light are mediated primarily by a subset of intrinsically photosensitive retinal ganglion cells (ipRGCs) expressing the photopigment, melanopsin (OPN4). These NIF functions include circadian entrainment, pupillary reflexes, and photic effects on sleep, mood, and cognition. We recently reported that mice of multiple genotypes exhibit reduced voluntary ethanol intake under both constant darkness (DD) and constant light (LL) relative to standard light-dark (LD) conditions. In the present study, we sought to determine whether these effects are mediated by melanopsin-expressing ipRGCs and their potential relationship to photic effects on the circadian system. To this end, we examined the effects of environmental lighting regimen on both ethanol intake and circadian activity rhythms in a genetically engineered mouse model ( Opn4aDTA/aDTA) in which melanopsin expression is completely blocked while ipRGCs are progressively ablated due to activation of attenuated diphtheria toxin A (aDTA) transgene under the control of the Opn4 promoter. As expected from previous studies, Opn4aDTA/aDTA mice displayed dramatic attenuation of circadian photosensitivity, but surprisingly, showed identical suppression of ethanol intake under both DD and LL as that seen in controls. These results demonstrate that the effects of lighting regimen on voluntary ethanol intake are independent of melanopsin-expressing ipRGCs and ipRGC-mediated photic effects on the circadian system. Rather, these effects are likely mediated by classical retinal photoreceptors and central pathways.

Reproductive management of the transitional mare

The mare is a seasonally polyoestrous long-day breeder with a physiological breeding season lasting from April–October in the Northern Hemisphere. The hypothalamic-pituitary-gonadal axis in the mare is subject to a circannual endogenous rhythm that is primarily regulated by day length. Increasing ambient photoperiod in the spring alters the pattern of melatonin secretion. The resulting stimulation of hypothalamic gonadotropin-releasing hormone secretion triggers pituitary follicular stimulating hormone release and follicular growth. Exposure of mares in deep anoestrus to a stimulatory photoperiod remains the most successful method of advancing the first ovulation of the season. The most commonly used lighting regimen is providing a fixed length of 15–16 hours of light exposure and 8–9 hours of dark, with a minimum light intensity in a stable of 100-lux (100–200 watt incandescent bulb). Other methods include using an additional 2.5 hours of light beginning at sunset and a pulse lighting system, providing 1 hour of light, 9.5–10.5 hours after the onset of darkness, during the photosensitive phase. Alternatively, the EquilumeTM light masks provide a unilateral LED light source emitting 50 lux of blue-light directly to the eye during the hours after dusk (until 11 pm). Mares that have not been maintained under lights, or that have been exposed to ineffective light therapy, may require therapeutic hormonal intervention to advance the onset of the first ovulation of the season. Many hormone protocols involving progestins, GnRH, dopamine agonists and recombinant luteinising hormone/follicle stimulating hormone have been studied with variable results. Therapy is typically more effective when started either in late transitional mares or following a period of stimulatory artificial photoperiod.

Reproductive management of the transitional mare

The mare is a seasonally polyoestrous long-day breeder with a physiological breeding season lasting from April–October in the Northern Hemisphere. The hypothalamic-pituitary-gonadal axis in the mare is subject to a circannual endogenous rhythm that is primarily regulated by day length. Increasing ambient photoperiod in the spring alters the pattern of melatonin secretion. The resulting stimulation of hypothalamic gonadotropin-releasing hormone secretion triggers pituitary follicular stimulating hormone release and follicular growth. Exposure of mares in deep anoestrus to a stimulatory photoperiod remains the most successful method of advancing the first ovulation of the season. The most commonly used lighting regimen is providing a fixed length of 15–16 hours of light exposure and 8–9 hours of dark, with a minimum light intensity in a stable of 100-lux (100–200 watt incandescent bulb). Other methods include using an additional 2.5 hours of light beginning at sunset and a pulse lighting system, providing 1 hour of light, 9.5–10.5 hours after the onset of darkness, during the photosensitive phase. Alternatively, the EquilumeTM light masks provide a unilateral LED light source emitting 50 lux of blue-light directly to the eye during the hours after dusk (until 11 pm). Mares that have not been maintained under lights, or that have been exposed to ineffective light therapy, may require therapeutic hormonal intervention to advance the onset of the first ovulation of the season. Many hormone protocols involving progestins, GnRH, dopamine agonists and recombinant luteinising hormone/follicle stimulating hormone have been studied with variable results. Therapy is typically more effective when started either in late transitional mares or following a period of stimulatory artificial photoperiod.

Temperature and Genotype Affect Anthocyanin Concentrations in Lettuce (Lactuca sativa)

In addition to their physiological and metabolic roles, anthocyanin (Antho) levels in lettuce contribute to visual and nutritional value-based assessments of crop quality. Although 7 genes are now thought to help regulate Antho synthesis, deposition and/or degradation in lettuce, the genetic and abiotic controls of Antho levels remain less well characterized in lettuce than other plants. Previous greenhouse studies demonstrated that Antho levels in diverse lettuce varieties are a function of temperature and lighting regimen. Here, three strongly related Lolla Rossa-type varieties (`Lotto', `Valeria', and `Impuls') varying in the number of genes controlling intensity of anthocyanins were subjected to differential temperature conditions in growth chambers to better discern the independent and interactive effects of temperature (T) and variety (V) on Antho levels. Fifteen day-old seedlings were placed into one of three chambers maintained at 20 °C day/night (D/N), 30 °C/20 °C D/N or 30 °C D/N. Antho levels were measured in leaf tissue collected 30 d after transplanting. The entire experiment was replicated twice. Although significant, the T x V interaction resulted from differences in the magnitude, not direction, of the change in Antho concentrations among varieties with changes in T. This suggests that T was a main driver of Antho levels in this study. Regardless of V, Antho concentrations were highest, moderate and lowest after growth at 20 °C D/N, 30 °C/20 °C D/N and 30 °C D/N, respectively. Likewise, regardless of T, Antho levels followed the pattern `Impuls' (three genes) > `Valeria' (two genes) > `Lotto' (one gene). Correlations among instrumented and human eye-based evaluations of color are also being tested in samples from both studies.

Production of inhibin A not B in rams: changes in plasma inhibin A during testis growth, and expression of inhibin/activin subunit mRNA and protein in adult testis

Previous studies have shown that changes in the plasma concentrations of immunoreactive inhibin measured by radioimmunoassay occur in parallel with growth and regression of the testes during a reproductive cycle in adult Soay rams induced by exposure to an artificial lighting regimen of alternating 16 week periods of long days and short days. With the development of new two-site ELISAs for sheep inhibin A and inhibin B, we have re-examined the relationship between FSH and dimeric, biologically active inhibin in the reproductive cycle in adult Soay rams. No signal was generated by sheep testicular extract, ram or ewe plasma, or sheep ovarian follicular fluid in the inhibin B ELISA. In contrast, ram plasma contained significant activity in the inhibin A ELISA, which diluted in parallel to the inhibin A standard, and was abolished by preincubation of ram plasma with monoclonal antibodies specific for the betaA, but not the betaB, subunit. These results indicate that the ram is the first adult male mammalian species identified to date in which the testes produce and secrete dimeric inhibin A and not inhibin B. Northern blot analysis and immunocytochemistry confirmed the presence of alpha, betaA and betaB inhibin/activin subunit mRNA and protein in the testes of adult rams. Changes in plasma inhibin A concentrations occurred in parallel with the growth and regression of the testes during the long day: short day: long day lighting regimen in adult Soay rams, confirming our previous observations with immunoreactive inhibin. During the growth phase of the testes in the first 8 weeks of exposure to short days there was a positive correlation between plasma FSH and inhibin A concentrations, indicating that during this phase the secretion of inhibin A is stimulated by FSH and that inhibin A did not act as a negative feedback hormone on FSH secretion. From week 8.5 to week 16.0 of exposure to short days, there was a negative correlation between FSH and testosterone concentrations, but not inhibin, indicating that when inhibin concentrations are high, testosterone acts as the negative regulator of FSH secretion. Thus, in intact adult rams, when the testes are fully active it appears that inhibin A may sensitize the pituitary to the negative feedback effects of testosterone, at which time they act synergistically to maintain plasma concentrations of FSH.

Flowering of Herbaceous Perennials under Various Night Interruption and Cyclic Lighting Treatments

Six long-day species of herbaceous perennials were grown under six night-interruption (NI) photoperiod treatments to determine their relative effectiveness at inducing flowering. Photoperiods were 9-hour natural days with NI provided by incandescent lamps during the middle of the dark period for the following durations: 0.5, 1, 2, or 4 hours; 6 minutes on, 54 minutes off for 4 hours (10% or 6/54 cyclic lighting); or 6 minutes on, 24 minutes off for 4 hours (20% or 6/24 cyclic lighting). For five species, the experiment was repeated with more mature plants; for the sixth, Rudbeckia fulgida Ait. `Goldsturm', following a cold treatment of 8 weeks at 5 °C. The species generally showed a quantitative flowering response to the NI duration until a saturation duration was reached; as the length of the uninterrupted night break increased, flowering percentage, uniformity, and number and plant height increased and time to flower decreased. Minimum saturation durations of NI were 1 hour for Coreopsis grandiflora Hogg ex Sweet `Early Sunrise' and Hibiscus moscheutos L. `Disco Belle Mixed', 2 hours for Campanula carpatica Jacq. `Blue Clips' and Coreopsis verticillata L. `Moonbeam', and 4 hours for unchilled R. fulgida `Goldsturm'. Echinacea purpurea Moench `Bravado' flowered similarly across all lighting treatments. The 6/24 cyclic lighting regimen induced flowering comparable to that under a continual 4-hour NI for four of the six species and the cold-treated R. fulgida `Goldsturm'. Flowering under the 6/54 regimen was generally incomplete, nonuniform, and delayed compared to that under saturation duration treatments. Three of five species flowered earlier when more mature plants were placed under the NI treatments. Cold-treated R. fulgida `Goldsturm' flowered more rapidly than unchilled plants and the saturation duration of NI decreased to 1 hour.