Supplemental Data

Files in this Data Supplement:

• supplemental material - Supplementary Figure 1. Microscopic visualization of nNOS catalytic activity using immunofluorescence to localize L-citrulline, a by-product of NO synthesis. Representative illustration of nNOS-expressing neurons (red) and L-citrulline detection (green) by fluorescent immunocytochemistry in coronal brain sections from diestrous (A) and proestrous (B) rats. V3, third ventricle; oc, optic chiasm. Right panels: high magnifications of the areas delineated by rectangles. Arrowheads indicate double-labeled neurons expressing nNOS and L-citrulline. Arrows indicate nNOS single-labeled neurons. Low magnification scale bar: 210 �m; high magnification scale bar: 60 �m.
• supplemental material - Supplementary Figure 2. Effect of pharmacological blockade of NO synthesis targeted to the preoptic region of the hypothalamus. L-NAME (5 mM), a NOS inhibitor, or vehicle (NaCl 0.9%) was delivered via a stereotaxically implanted stainless steel cannula connected to a subcutaneously placed osmotic pump delivering its content at a rate of 0.5 �l/hr for 7 days. A. Disruption of estrous cyclicity in young adult rats by the infusion of L-NAME into the preoptic region. Infusion starts at P0 (arrow pointing up) and ends 7d later (arrow pointing down), when the pump content is exhausted. The estrous cycle profile of representative animals are depicted. Di, Diestrus; Pro, Proestrus; Es, estrus. B. Analysis of the alterations of estrous cyclicity caused by L-NAME in the preoptic region of the hypothalamus. Animals infused with L-NAME in the preoptic region stayed a significantly greater percentage of days in diestrus when compared to NaCl-infused animals (p = 0.001, t-test). Error bars indicate SEM. Numbers at the bottom of each bar graph are numbers of independent observations per group.