Mammary localization and abundance of laminin, fibronectin, and collagen IV proteins in prepubertal heifers

The objective was to determine localization and abundance of extracellular matrix proteins fibronectin, laminin, and collagen in mammary tissues from ovariectomized or intact prepubertal heifers. Mammary parenchyma and fat pad tissues were collected from 14 6-mo-old heifers: eight were ovariectomized between 1 to 3 mo of age, and six were used as intact controls. Distribution of total collagen was assessed by Sirius Red staining of tissue sections. Fibronectin, laminin, and type IV collagen were assessed by immunohistochemistry. Abundance of fibronectin and laminin was also analyzed by western blotting. Total mammary mass was much less in ovariectomized animals (130 +/- 21 vs. 304 +/- 25 g). Histological structure differed as parenchyma from intact animals contained abundant, complex branching epithelial terminal ductular units, whereas terminal ductular units from ovariectomized animals were mostly major ductal structures with little or no branching. Collagen fibers were abundant and densely packed throughout interlobular stroma and were less abundant and more diffuse within intralobular stroma. Type IV collagen was primarily in basal lamina of mature ducts, whereas fibronectin and laminin staining were present throughout parenchymal stroma, in both intact and ovariectomized animals. Using western blotting, fibronectin was more abundant within parenchyma than in the fat pad and significantly higher in parenchyma from ovariectomized heifers. Laminin was more abundant in parenchyma from intact than ovariectomized animals (30 vs. 17 densitometric units/mg of tissue), but laminin was similar between parenchyma and fat pad. These results provide initial evidence that fibronectin, laminin, and collagen participate in regulation of heifer prepubertal mammary development.