Substrain and light regime effects on integrated anxiety-related behavioral z-scores in male C57BL/6 mice-Hypomagnesaemia has only a small effect on avoidance behavior
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Updated: Mar 24, 2026

Investigating Migraine-Like Behavior Using Light Aversion in Mice
Published on: August 11, 2021
M Labots1, X Zheng2, G Moattari2
1Division of Animal Welfare & Laboratory Animal Science, Department of Animals in Science and Society, Faculty of Veterinary Medicine, Utrecht University, P.O. Box 80166, 3508 TD Utrecht, The Netherlands; Brain Center Rudolf Magnus, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands.
Abstract:
Magnesium (Mg) has been described to possess an anxiolytic function, but a number of studies present inconsistent results on this matter. In this study the effect of Mg deficiency on anxiety-related behavior, brain and blood plasma Mg in young adult male C57BL/6JOlaHsd and C57BL/6NCrl mice was studied. The animals were put on a control or Mg deficient diet from day 0 and significant hypomagnesaemia was evident from day 12 onwards in the test animals. Housing and test conditions were under either conventional light regime (white light behavioral test conditions) or reverse light regime (red light behavioral test conditions). The animals were tested in three tests for unconditioned anxiety: the modified Hole Board (day 14), the light-dark test (day 21) and the elevated plus maze (day 28). Overall integrated behavioral z-scores were calculated over these three behavioral tests. Mg showed a structure dependent distribution at the level of the brain, that differed between C57BL/6 substrain and light regime (conventional versus reverse), respectively. Likewise, total brain Mg did differ between substrain and light regime, but was not affected by the diet. Animals on the Mg deficient diet housed under conventional light regime had a higher final (day 28) blood plasma corticosterone level as compared to controls. Animals housed under reverse light regime exhibited no diet effect of plasma corticosterone levels. The significant hypomagnesaemia at blood plasma level resulted in an effect of Mg deficiency on avoidance, but not overall anxiety-related behavior. Significant differences regarding avoidance behavior were found between the two substrains and light regimes, respectively.
Insights
Magnesium deficiency in mice affected avoidance behavior, not overall anxiety. Diet and light conditions influenced brain magnesium distribution and stress hormone levels.
Area of Science:
- Neuroscience
- Behavioral Science
- Nutritional Science
Background:
- Magnesium's anxiolytic effects are debated due to inconsistent study results.
- Understanding magnesium's role in anxiety requires investigating its impact on behavior and neurochemistry.
Purpose of the Study:
- To investigate the effects of magnesium deficiency on anxiety-related behaviors in mice.
- To examine brain and blood plasma magnesium levels under different housing and light conditions.
- To assess the influence of magnesium deficiency on stress responses.
Main Methods:
- Young adult male C57BL/6 mice were fed control or magnesium-deficient diets.
- Animals were housed under conventional or reverse light regimes.
- Anxiety-related behaviors were assessed using the Hole Board, light-dark test, and elevated plus maze.
- Brain and blood plasma magnesium levels, and corticosterone were measured.
Main Results:
- Magnesium deficiency induced hypomagnesemia, affecting avoidance behavior but not overall anxiety.
- Brain magnesium distribution varied by mouse substrain and light regime, independent of diet.
- Animals on a magnesium-deficient diet under conventional light showed elevated corticosterone.
- Reverse light housing negated the diet's effect on corticosterone.
Conclusions:
- Magnesium deficiency specifically impacts avoidance behavior, suggesting a nuanced role in anxiety.
- Environmental factors like light regime significantly modulate magnesium's effects on neurochemistry and stress.
- Further research is needed to clarify magnesium's complex interactions with behavior and physiology.

