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neurotoxicity of these drugs on the developing nervous system, when administered alone or in combination. Methods: Newborn rats were given a subcutaneous single injection on day 3 of life (P3) of either: 1) saline, 2) caffeine (100mg/kg), 3) morphine (10mg/kg) or 4) both medications combined. Twenty-four hours post-injection rats were euthanized and the brains were quick frozen in iso-pentane. Sections were stained using Fluoro-jade B to identify and quantify neuronal death in specific brain areas. Positive staining cells in the cortex, caudate/putamen, nucleus accumbens, globus pallidus, thalamus, hypothalamus, superior colliculus and hippocampus were counted, and the extent of cell death determined per mm 3 . Results: Weight gain in those rat pups receiving both caffeine and morphine was significantly less over the 24 hours postinjection than was the control group, or in those rats receiving either of the drugs alone (pϽ0.05). The neocortical regions did not show any significant increase in cell death. Morphine alone, did not cause an increase in cell death in any of the regions examined. Caffeine alone resulted in a significant increase in cell death, compared to control, in all areas examined (pϽ0.001), with the exception of the thalamus and caudate/ putamen. In combination, a significant increase in cell death was seen over control in the hypothalamus, the thalamus and the hippocampus (pϽ0.01). Conclusions: Medications commonly utilized in the treatment of premature infants may not be benign. Caffeine in particular causes an increase in cell death. The ramifications of these drugs long term on brain development and behavior need further investigation.
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