Differential expression of estrogen receptor and neuropeptide Y by brainstem A1 and A2 noradrenaline neurons

The release of noradrenaline and neuropeptide Y appears to be regulated by estrogen in a co-ordinated fashion within specific brain regions. The present study has used double and triple-labelling immunocytochemical procedures to determine the patterns of nuclear estrogen receptor and neuropeptide Y expression by brainstem A1 and A2 noradrenergic neurons in the female rat. Estrogen receptor-immunoreactive cells were detected within the ventrolateral medulla, nucleus tractus solitarius, area postrema and, in the very caudal medulla, the reticular nuclei and spinal nucleus of the trigeminal nerve. Cells double labelled for the estrogen receptor and dopamine-beta-hydroxylase were identified in largest numbers (up to seven double-labelled cells per 30-microm-thick coronal section) in the caudal-most medulla, where approximately 30% of A1 and 60% of A2 neurons were immunoreactive for the estrogen receptor. These percentages reduced in a linear fashion in more rostral sections and at the level of the area postrema, no co-expression was evident in the ventrolateral medulla and only 10% of A2 neurons displayed estrogen receptor immunoreactivity. Fluorescence double-labelling studies undertaken in colchicine-treated rats revealed that 50% and 90-100% of tyrosine hydroxylase-immunoreactive cells were positive for neuropeptide Y in the rostral ventrolateral medulla and nucleus tractus solitarius (up to 15 double-labelled cells per section), respectively. This pattern of co-expression also showed a rostrocaudal bias, but in the opposite direction, such that none of the caudal-most A1 and only 10% of caudal A2 neurons were immunoreactive for neuropeptide Y. Triple-labelling experiments revealed the presence of a total of only three triple-labelled cells in the ventrolateral medulla and none in the nucleus tractus solitarius of four rats. Double-labelling studies examining estrogen receptor and neuropeptide Y co-expression similarly found only three double-labelled cells in the ventrolateral medulla. These findings provide immunocytochemical evidence for a clear rostrocaudal topography in nuclear estrogen receptor synthesis by A1 and A2 neurons and show a reverse rostrocaudal bias in neuropeptide Y expression by these cells. The absence of any substantial neuropeptide Y and estrogen receptor co-expression in A1 and A2 neurons indicates that these two proteins are very likely to be differentially expressed by brainstem noradrenergic neurons. Such observations provide further evidence for the biosynthetic and functional heterogeneity of brainstem noradrenergic cells and suggest that A1 and A2 neurons transmitting information on estrogen status within the brain are unlikely to utilize neuropeptide Y as a co-transmitter.