Mouse strain-specific patterns of mammary epithelial ductal side branching are elicited by stromal factors

Variations in mammary ductal side branching patterns are known to occur between different strains of mice and this is related to the rate of spontaneous mammary cancers, which are increased in those strains which show highly side-branched mammary architecture. The cause of the variation in ductal side branching between mouse strains is unknown, but epithelial, stromal, and endocrine factors have been implicated. To define the mammary elements responsible for controlling strain-specific ductal side branching patterns, we formed recombined mammary glands from epithelial and stroma elements taken from highly side-branched 129 and poorly side-branched C57BL/6J mammary glands and transplanted them to Rag1(-/-) hosts on the inbred C57BL/6J background. When 129 epithelium was recombined with C57BL/6J stroma the poorly side-branched C57BL/6J pattern was observed. C57BL/6J epithelium recombined with 129 stroma resulted in development of the highly side- branched pattern, as did 129 epithelium recombined with 129 stroma. All transplants used the same C57BL/6J endocrine background, demonstrating that strain differences in the mammary stroma are responsible for the strain-specific ductal side branching patterns and that strain differences in epithelium or endocrine background play no part. Genes currently known to influence side branching by means of the stroma include activin/inhibin, epidermal growth factor receptor (EGFR), Wnt-2, Wnt-5a, and Wnt-6. Of these, Wnt-5a mRNA expression was decreased in 129 mammary glands compared with C57BL/6J mammary glands, but in F2 129:C57BL/6J animals Wnt-5a mRNA expression level did not correlate with the highly variable side branching patterns observed. These experiments exclude variation in the expression level of known candidate genes as the mechanism responsible. Regardless of underlying mechanism, transplantation without regard to the genetic background of the stromal donor, whether inbred or mixed, will compromise experiments with side branching and associated gene expression endpoints.