Immunohistochemical localisation of the 90, 70 and 25 kDa heat shock proteins in control and caffeine treated rat embryos

Human and animal experimental data demonstrate that in utero exposure to caffeine results in intrauterine growth retardation and long-term behavioural and reproductive effects. We have suggested that the disruption of normal transcription and translation associated with the initiation of the heat shock response may be a possible mechanism of action of caffeine. This hypothesis was investigated using immunohistochemistry to determine whether an acute (3 h) dose of 30 mg/kg caffeine alters the distribution of hsp 90, 70 and 25 in 10.5–12.5 g.d. rat embryos. In the control embryos hsps 90 and 70 were distributed throughout the embryo with no areas of specific accumulation. Hsp 25 was localised to the developing myocardium of 10.5, 11.5 and 12.5 g.d. embryos and the myotome of 11.5 and 12.5 g.d. embryos. The appearance of hsp 25 was correlated with the onset of muscle fibre differentiation and it is suggested that hsp 25 is associated with cytoskeletal proteins. Following dosing with caffeine no change in the distribution of staining for hsp 90, 70 and 25 was found. These results strongly suggest that caffeine's mechanism of action does not involve initiation of the heat shock response.