Formalin-induced nociceptive behavior and c-Fos expression in middle-aged female rats

The impact of the estrous cycle on the nociceptive response in middle-aged female rats was assessed using the formalin test and c-Fos immunoreactivity as a marker of neural activation. Young (2-month-old) and middle-aged (11-month-old) rats were examined, dividing the middle-aged rats into two groups based on their estrous cycle: regular 4-day estrous cycle and irregular estrous cycle. The right hind paw was subcutaneously injected with 50microl of 2% formalin or saline as a control. Behavioral changes were observed for 1h. Cycling rats were used during proestrus. Middle-aged female rats had a significantly higher score for nociceptive behavior compared to young rats, irrespective of estrous cyclicity, which suggests that aging, not the ability to maintain estrous cyclicity, causes hypersensitivity to the formalin injection. Immunohistochemical analysis found that the brain response to formalin injection was also more sensitive in middle-aged rats than young rats; a significant increase in the number of c-Fos immunoreactive cells was found in the ventral portion of the lateral septum of middle-aged rats injected with formalin compared to young and middle-aged rats injected with saline, irrespective of estrous cyclicity. Based on these results, we conclude that the sensitivity to painful stimuli in middle-aged female rats, which are in a neuroendocrine state similar to pre- and peri-menopausal women, is associated with age and not affected by reproductive ability.