Physiology & Behavior 266 (2023) 114186

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Physiology & Behavior

journal homepage: www.elsevier.com/locate/physbeh

The role of daylight exposure on body mass in male mice

O.Hecmarie Meléndez-Fernández a, *, James C. Walton a, A.Courtney DeVries a, b, c,
Randy J. Nelson a
a
Department of Neuroscience, Rockefeller Neuroscience Institute, Morgantown, WV 26505 USA
b
Department of Medicine, Division of Oncology/Hematology, Morgantown, WV 26505 USA
c
West Virginia University Cancer Institute, West Virginia University, Morgantown, WV 26505, USA

A R T I C L E I N F O A B S T R A C T

Keywords: Physiology and behavior are synchronized to the external environment by endogenous circadian rhythms that
Circadian rhythms are set to precisely 24 h by exposure to bright light early in the day. Exposure to artificial light outside of the
Light typical solar day, such as during the night, may impair aspects of physiology and behavior in human and non-
Body mass
human animals. Both the intensity and the wavelength of light are important in mediating these effects. The
Metabolism
present report is the result of an unplanned change in our vivarium lighting conditions, which led to the
Artificial light at night
observation that dim light during the daytime affects body mass similarly to dim nighttime light exposure in male
Swiss Webster mice. Mice exposed to bright days (≥125 lux) with dark nights (0 lux) gained significantly less
weight than those exposed to bright days with dim light at night (5 lux) or dim days (≤60 lux) with either dark
nights or dim light at night. Notably, among the mice exposed to dim daytime light, no weight gain differences
were observed between dark nights and dim light at night exposure; however dim light at night exposure shifted
food intake to the inactive phase as previously reported. The mechanisms mediating these effects remain un­
specified, but it appears that dimly illuminated days may have similar adverse metabolic effects as exposure to
artificial light at night.

1. Introduction
Physiology and behavior have endogenous circadian rhythms with a
period of about 24 h and are synchronized to the external solar day via
light and other synchronizing cues. In mammals, information about the
daily light-dark cycles is conveyed to the suprachiasmatic nucleus (SCN)
of the hypothalamus by direct photic input from the retinal ganglion
cells. Circadian rhythms in the SCN and other tissues reflect a molecular
clock based on transcriptional translational feedback loops at the
cellular level comprising core clock genes, Clock, Bmal1, Per, and Cry
among other supporting clock genes [1,2]. The duration and intensity of
light as a synchronizer (zeitgeber) plays a key role in entraining the
molecular clock to the time of day. Prolonged or inappropriate exposure
to light into the biological night has deleterious effects on physiology
and behavior [3,4].
We and others have reported that disrupted circadian rhythms by
exposure to light at night in nocturnal rodents increases body mass
(reviewed in [5]). Exposure to dim light at night specifically attenuates
the rhythm in Per1 and Per2 gene and protein expression in the SCN
around the time of the light-dark transition [4,6]. Furthermore, rhyth­
mic expression of Bmal1, Per1, Per2, Cry1, Cry2, and Rev-Erb are all
suppressed in the liver by exposure to dim light at night [4]. Nocturnal
light exposure may affect the liver through both autonomic and hor­
monal pathways [7]. These observed changes in rhythmic clock gene
expression associated with exposure to dim light at night are not suffi­
cient to disrupt either the glucocorticoid rhythm or locomotor activity
rhythm [4,8]. Much of the change in metabolism leading to dim artificial
light at night (ALAN)-induced weight gain is attributed to adjustments in
the timing of food intake [8–10].
During a recent study to examine cardiometabolic effects of dis­
rupted circadian rhythms by light at night, we noted, in our second
cohort of mice, that those housed in ALAN did not gain mass relative to
their counterparts housed in dark nights, as those in the first study
cohort did, and what was predicted based on previous work [4,8].
Further investigation revealed that although the nighttime light expo­
sure was carefully controlled in our experiment, daytime lighting
throughout the vivarium had been altered between cohorts and that
these animals were exposed to relatively lower levels of light during the

* Corresponding author at: 108 Biomedical Road, BMRC Room 370, Morgantown, WV 26506, USA.
E-mail address: ohm0001@mix.wvu.edu (O.Hecmarie Meléndez-Fernández).

https://doi.org/10.1016/j.physbeh.2023.114186
Received 21 February 2023; Received in revised form 25 March 2023; Accepted 4 April 2023
Available online 5 April 2023
0031-9384/Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
O.Hecmarie Meléndez-Fernández et al. Physiology & Behavior 266 (2023) 114186

day compared to the first cohort of the study. We took advantage of this
serendipitous discovery to retrospectively test the hypothesis that day­
time light exposure affects metabolism in male mice.
2. Methods
Animals and Lighting Conditions. Sixty-four adult (7 weeks of age)
male Swiss Webster mice were obtained from Charles River Labs (Wil­
mington, MA, USA) for this study. Upon arrival all mice were main­
tained in a 14:10 light:dark cycle (LD), and allowed to acclimate to local
conditions for one week. Mice were group housed (4/cage) in clear
polysulfone plastic cages (Allentown LLC, NJ, USA), dimensions 19.37
cm x 18.06 cm x 39.83 cm, with corncob bedding and a paper nestlet
puck for enrichment. The ambient temperature was maintained at 22
±2 ◦ C, and mice were provided with Teklad 2018 chow (Envigo, Mad­
ison, WI) and filtered tap water ad libitum. After one week, mice were
randomly assigned to one of two conditions as follows: standard con­
ditions LD; 125 lux at cage level during the day, 0 lux at night; or dim
artificial light at night (ALAN; 125 lux during the day, 5 lux at night), for
eight weeks (n = 32/light condition, in 2 cohorts per condition; 16 per
cohort, per condition). Following assignment to experimental group,
mice were singly housed and cages were placed on static racks, equi­
distant from a set point, ~2 in from the rack’s edge (LD) or an LED light
strip (ALAN) and nighttime light levels were measured inside the cage
with a Mavolux 5032C illuminance meter (Nürnberg, Germany). To
measure illuminance at the cage level, the light meter sensor was placed
in the center of the cage with the sensors facing toward the ceiling. Low-
level ALAN was supplied by LUMA5 LED light strips (Hitlights Inc.; 1.5
W/ft, 5000 K “cool white”, 1200 lm) and output was set to 5 lux via
rheostat control of the light strip. Throughout the experiment, mice were
weighed each week between zeitgeber time, (ZT) 10–12. Four weeks into
the second cohort of the experiment, we noted that there was not a
differential in body mass between the two light treatments in the second
cohort. At this time, we noted that daytime lighting conditions were
~60 lux at cage level, dimmer than the standard housing conditions
(>125 lux), as measured by our luxmeter. Thus, we decided to collect
food consumption data, which was not part of the original protocol for
this study, and conclude the study prematurely. Food consumption was
measured over 24 h during the last experimental week, for mice in this
(second) cohort. Because we prematurely ended data collection for

Fig. 1. Experimental timeline. Planned experimental timeline for cardiometabolic study consisted of 8 weeks of experimental light exposure with weekly body
mass measurements (A). The study would be completed in two cohorts consisting of two experimental light conditions, standard conditions (LD; 125:0 lux), and
bright days with dim artificial light at night (ALAN; 125:5 lux) with a total of n = 32 / light condition. However, vivarium light conditions were unexpectedly altered
(B), producing dim daytime illumination at cage level (~60 lux). We took advantage of this serendipitous discovery to retrospectively test the hypothesis that
daytime light exposure affects metabolism in male mice. Thus, we decided to collect food consumption data, which was not part of the original protocol for this study,
and conclude the study prematurely.

2
O.Hecmarie Meléndez-Fernández et al.
cohort 2 due to the unplanned lighting alteration, the data presented
here for cohort 1 are truncated at week 4 for comparison purposes with
cohort 2 (see Fig. 1 for experimental timeline). Upon conclusion of the
study for both cohorts, mice were deeply anesthetized with a lethal
overdose of Euthasol (Patterson Companies, MN, USA), and decapitated.
All experimental procedures were approved by West Virginia University
Institutional Animal Care and Use Committee, and animals were main­
tained in accordance with the guidelines established by the National
Institutes of Health in the Guide for the Care and Use of Laboratory
Animals.
Statistical analyses. Outliers were determined using the Grubb’s test
and were removed a priori. Body mass data were analyzed using
repeated measures ANOVA. Average daily food intake data were
analyzed using a Student’s t-test. Phase-specific food intake data were
analyzed initially using a within lighting condition paired t-test, fol­
lowed by a one-way ANOVA. Significant ANOVAs were further analyzed
with a Fisher’s LSD post hoc comparison. In all cases, differences between
group means were considered statistically significant if p ≤ 0.05. All
statistical analyses were performed using SPSS v28 and figures were
generated using GraphPad Prism version 9.5.
3. Results
Body mass. As depicted in Fig. 2, mice in all lighting conditions
gained weight during the experiment (body mass, Fig. 2A; F(1.928, 111.4)
= 239.9, p < 0.001), and as an overall percent change from baseline
(Fig. 2B; F(4, 216) = 276.028, p < 0.001), however, the different lighting
conditions affected weight gain (F(3, 54) = 4.041, p = 0.012). Mice
exposed to the brightest daytime light combined with dark nights
(125:0 lux) had significantly lower weight gain (p < 0.05) than their
counterparts exposed to bright days with dim light at night (125:5 lux),
dim days and dark nights (60:0 lux), or mice in dim days and dim light at
night (60:5 lux), which did not differ from one another (p > 0.05).
Food consumption. To determine whether dim light at night exposure
altered food consumption or the timing of food intake in mice exposed to
dim daylight (60 lux; second cohort), we measured chow consumption
at phase transitions in the final week of the study. Under dim daylight
conditions, average daily food consumption did not differ between mice
exposed to dark nights and those exposed to dim light at night (t(17) =
0.5542, p = 0.5867, Fig. 3A). Regardless of nighttime lighting condition,
mice consumed more food during their active phase than their inactive
phase (p < 0.05, Fig. 3B). However, both phase and nighttime lighting
Fig. 2. Effects of daytime light intensity on body mass gain in male Swiss Webster mice exposed to dim light at night. Mice exposed to bright days (125 lux)
and dark nights (0 lux) gain significantly less body mass (absolute mass, A; percent change, B) than mice exposed to bright days with dim light at night (125:5 lux),
dim days and dark nights (60:0 lux), or mice in dim days and dim light at night (60:5 lux). All data shown as mean ± SEM; *p ≤ 0.05.
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Physiology & Behavior 266 (2023) 114186
condition affected food consumption (F(3, 39) = 50.538, p < 0.001)
where dim light at night exposure reduced food consumption in the
active phase, with a concurrent increase in consumption during the
inactive phase (p > 0.05, Fig. 3B).
4. Discussion
Due to a change in daytime vivarium lighting during the course of an
ongoing study, we made the observation that, irrespective of the
nighttime light exposure, dim light during the daytime may increase
body mass similar to what has been observed in mice exposed to dim
light at night. Exposure to dim days and nights (60:5 lux), elevated food
intake during the inactive phase relative to the dim day-dark night-
housed mice (Fig 2). However, because we were unable to determine
food consumption data for the bright day-dark night (125:0 lux) cohort,
the question remains whether exposure to dim days, in common with
exposure to dimly lit nights, significantly elevates inactive-phase food
intake relative to dark night conditions (i.e., bright days and dark nights;
≥125:0 lux), which is a limitation of our study. Further, we recognize
that a one-time snapshot of food consumption data (at week 4) may not
necessarily be reflective of eating patterns for the prior weeks. This
would require follow up experiments to test the hypothesis that dark
nights bright days provide a distinct day-night demarcation, which al­
lows mice to sustain distinct diel rhythms of activity and food con­
sumption across exposure to dim nights or dim days. Nonetheless, this
serendipitous finding may suggest that although disruption of circadian
rhythms by dim nocturnal light exposure is a factor driving changes in
metabolism leading to altered timing of food intake and elevated weight
gain in mice, dim illumination during the day can also increase weight
gain. The mechanisms mediating this alteration, remain to be eluci­
dated. As noted above, exposure to dim light at night disrupts molecular
circadian rhythms and leads to adverse outcomes [4,8]. Data from night
shift workers demonstrate that exposure to artificial nocturnal illumi­
nation elevates body mass, at least partly, by shifting the time during
which workers consume the majority of their daily calories [11], and by
altering metabolic processing through elevation in postprandial plasma
glucose [12]. These data are consistent with the rodent literature on the
effects of dim light at night [4,8]. However, it is also the case that
circadian rhythms and metabolic outcomes are strongly influenced by
the intensity of light during the day. For instance, in humans, early
morning bright light therapy (at the beginning of the active phase)
decreased body mass index and elevated total fat loss [13,14], likely
O.Hecmarie Meléndez-Fernández et al.
Fig. 3. Effects of day and night intensity of light exposure on daily caloric intake in male Swiss Webster mice. Mice housed in dim daylight (60 lux) consumed
the same amount of chow independent of light at night exposure (A). However, independent of changes in body mass (see Fig 1), exposure to dim ALAN (5 lux)
elevates caloric consumption during the inactive phase and decreases intake during the active phase (B). All data shown as mean ± SEM; *p ≤ 0.05.
through decreased appetite, (self-reported by participants), and conse­
quently, reduced food intake [13]. Additionally, exposure to bright light
in the morning increases energy expenditure through elevation of oxy­
gen consumption [15], and by stimulating sympathetic activation [16,
17], which could also mediate the effects on body mass. Thus, the lack of
bright light exposure during the day in the current study may have had
disruptive effects on the circadian clock similar to what is observed with
dim light exposure at night, leading to increased body mass
accumulation.
Recent work has begun to highlight the importance of strong daytime
light signals in maintaining circadian rhythmicity and health. An elegant
study recently demonstrated that duration, intensity, and wavelength of
light exposure during the day directly affects physiology and meta­
bolism in mice, with the most robust circadian rhythmicity and health
being observed in mice exposed to the strongest entraining signal during
the day (blue-enriched light) [18]. The positive effects of bright daytime
illumination compared to dim days have also been observed in diurnal
rodents. In diurnal four-striped mice, daytime light intensity is directly
and positively correlated with circadian rhythm amplitude of SCN ac­
tivity, body temperature, and activity [19]. In diurnal fat sand rats,
compared to dimmer daytime light (800 lux), daytime bright light
(3000 lux) exposure enhanced Per2 rhythmicity in the SCN and liver
concurrent with reduced body mass and lower fasting and post-glucose
tolerance test blood glucose concentrations [20]. A recent study aimed
at assessing the interaction of daytime light intensity and circadian
disruption under different diets found that higher intensities of daytime
light (169 lux compared to 78 lux) were necessary to observe the effects
of circadian disruption on metabolism and weight gain in mice fed a low
fat diet [21]. Notably, when fed a high fat diet the combined effects
changed, where daytime light intensity and circadian disruption inter­
acted to affect metabolism and weight gain, and in both diets the effects
of light intensity and circadian disruption appeared to be uncoupled
from timing of food intake [21].
In conclusion, our serendipitous findings add to the growing body of
literature supporting the critical importance of circadian rhythm hy­
giene in physiology, metabolism, and health. Taken together, work in
this field highlights the biological importance of not only limiting
exposure to light at night, but also having appropriate exposure to the
optimal intensity of light during the daytime to optimize circadian
rhythmicity. Future studies will be necessary to tease apart the impor­
tance of the interrelationships among the circadian amplitude of light:
dark signals, the molecular circadian clock, diet composition, and
feeding rhythms in the maintenance of health, with obvious implications
for incorporating chronotype in personalized medicine.
4
Physiology & Behavior 266 (2023) 114186
Funding
This work was supported by the National Institutes of Health grant #
5RO1NS092388 to RJN and ACD. OHMF was supported by the NIH
National Institutes of General Medical Sciences predoctoral fellowship
T32 GM132494. The content is solely the responsibility of the authors
and does not necessarily represent the official views of the US National
Institutes of Health.
Declaration of Competing Interest
The authors have no conflict of interest to disclose.
Data availability
Data will be made available on request.
Acknowledgements
We thank William H. Walker II for valuable feedback in the prepa­
ration of this manuscript, and Terri Poling and Amanda Rack for assis­
tance with animal care and husbandry.
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