Caffeine is a common medication benefitting apnea of prematurity (AOP). Despite this, controversy remains regarding its long-term effects. This study determines long-term neuropathological and neurobehavioral effects of clinically relevant caffeine doses in immature rat brains.
Methods:
Rat pups received subcutaneous injections of 20 mg/kg caffeine citrate on P3, and 15 mg/kg daily from P4 to P7. Control pups received identical volumes of normal saline. During P4–P21, rat pups underwent early neurobehavioral reflex testing. An object recognition test on P63 and the Morris water maze test on P160 assessed long-term neurobehavioral effects. Rats were euthanized on P4, P8, P21, and P160 and their brains were examined for indications of cell death. Volumetric measurements were taken of the hippocampus, regionally, and the cerebral cortex.
Results:
Fluoro-Jade B stain analysis on P4 showed significant, although transient, caffeine-induced neuronal degeneration in the nucleus accumbens, globus pallidus, substantia nigra, thalamus, hypothalamus, and the hippocampal CA1. Anti-NeuN immunohistochemistry on P21, but not on P160, revealed significant neuronal loss in the hippocampal CA1, and hypothalamus in the caffeine group. Axonal formation (Anti-Neurofilament-M immunohistochemistry), and volumetric and neurobehavioral measures revealed no difference between control and caffeine groups.
Conclusion:
Although neuropathological differences were transiently noted, caffeine use analogous to AOP treatment had no long-lasting neuropathological or neurobehavioral effects on the newborn rat pups. These findings indicate the relative safety of caffeine in normal newborn brains. Further studies need to determine subtle changes in brain connectivity and adenosine receptor development.
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