The effects of age and estradiol on hypothalamic content and distribution of tubulin and calmodulin were examined by radioimmunoassay in ovariectomized C57BL/6J female mice. A small (14%) increase in particulate tubulin, but not soluble tubulin, was found in the hypothalamus of reproductively senescent mice (20 months) compared to young (8 months) controls. This alteration was limited to tubulin; calmodulin content was unaffected by age. Post-castration serum LH levels were lower in old, ovariectomized controls relative to young controls, but physiologic levels of estradiol, achieved by subcutaneous implants, suppressed LH levels in both age groups. In contrast to LH and uterine weight, hypothalamic tubulin and calmodulin were unaffected by estradiol treatment. These results suggest that the negative feedback effect of estradiol on LH secretion is exerted by a mechanism other than redistribution of hypothalamic tubulin or calmodulin, or that changes are restricted to a discrete sub-population of neurons.