A strontium and neodymium isotopic investigation of plagioclase-rich cumulate rocks in the 1.43 Ga Laramie anorthosite complex (LAC) provides new insights into the evolution of magma chambers and underlying magma plumbing systems during the crystallization of Proterozoic anorthosites. The 725 km2 LAC intruded the boundary between Archean rocks of the Wyoming Province in the north and Proterozoic island arc terranes in the south, the Cheyenne belt, at pressures of 3–4 kilobars. Initial strontium and εND isotopic ratios from the three large composite anorthositic intrusions (Poe Mountain, Chugwater, and Snow Creek) and late troctolitic intrusions range from 0.7034–0.7055 and +0.9 to −4.9, respectively. The isotopic ratios in the cumulates vary as a result of (1) the isotopic composition of the parental mantle-derived high-Al gabbroic magmas, (2) the amount of contamination by Archean rocks during ascent through the crust, and (3) mixing of replenishing magmas and resident magmas with different isotopic compositions in the magma chamber at the final level of emplacement.Strontium isotopic and normative An variations across the 5–7 km thick stratigraphic section of layered plagioclase-rich cumulates in the 200 km2 Poe Mountain anorthosite indicate that multiple inputs of magma are needed to construct even relatively small intrusions in Proterozoic anorthosites. The earliest and stratigraphically lowest intrusion was emplaced as a plagioclase-rich magma that crystallized large volumes of compositionally homogeneous anorthosite. At least three subsequent chamber-wide episodes of magma replenishment, followed by mixing, are indicated by abrupt shifts in ISr and/or normative An in the stratigraphically higher levels of the Poe Mountain anorthosite. Strontium and neodymium isotopic disequilibrium between a high-Al clinopyroxene megacryst and the layered cumulates is consistent with a high-pressure origin for Al-rich pyroxene megacrysts in Proterozoic anorthosites. The megacryst crystallized in a magma chamber at pressures of 10–12 kilobars from relatively uncontaminated basaltic parent magma (high-Al gabbro) and was transported through the crust in progressively contaminated anorthositic magma.The range of observed isotopic compositions in the anorthositic rocks is nearly identical to that of high-Al gabbroic dikes in the LAC, supporting the proposition that high-Al gabbros are parental to anorthosite (Mitchell et al., 1995). The isotopic data require that the anorthositic parental magmas were contaminated during ascent through the crust by Archean orthogneisses and/or metapelitic rocks. Decreasing ISr and increasing εNd with a relative decrease in age of the intrusions (Snow Creek → Poe Mountain/Chugwater → troctolites) indicates that the magma conduit became increasingly insulated from crustal contamination over time.This study indicates that the strontium and neodymium isotopic systems can be used to distinguish between processes that occurred at lower crustal/upper mantle pressures, during the ascent of magma diapirs, and within individual magma chambers in Proterozoic anorthosites, and underscores the need for stratigraphic control when addressing the origin of plagioclase-rich cumulates.