title: Hormone-inducible casein messenger RNA in a serum-free organ culture of whole mammary gland.
abstract: The whole second thoracic mammary gland of estradiol-17beta + progesterone primed 3- to 4-week-old BALB/c female mice was induced to pregnancy-like lobulo-alveolar morphogenesis after 6-day cultivation in a serum-free culture medium containing a "growth promoting" hormone mixture, insulin + prolactin + growth hormone (somatotropin) + estradiol + progesterone. No radioimmunologically detectable casein was present in these glands. Subsequent cultivation for another 6 days in a "lactogenic" medium with the hormones insulin + prolactin + cortisol produced abundant milk-like secretory material in the alveolar lumen. RNA of the mammary gland after estradiol + progesterone priming or cultivation in the "growth-promoting" medium failed to show a measurable amount of casein mRNA activity when assayed in a cell-free protein synthesis system derived from Ehrlich ascites ribosomes, rabbit reticulocyte factors, and tRNA. However, the glands sequentially cultivated in the "growth-promoting" and the "lactogenic" media showed a high level of casein mRNA activity in the heterologous cell-free protein synthesis system. Sodium dodecyl sulfate/polyacrylamide gel electrophoretic characteristics of the immunoprecipitable (by antibody to mouse milk casein) polypeptides directed by the mammary RNA induced in organ culture medium containing the lactogenic hormones were similar to the characteristics of the polypeptides directed by mammary polysomes of lactating mice. These results demonstrate hormonal induction of a specific mRNA in a sequential two-step culture of an entire organ in a serum-free chemically defined medium.
mesh_terms: D000818:Animals; D002364:Caseins / Q000096:biosynthesis*; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D013006:Growth Hormone / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008321:Mammary Glands, Animal / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D009924:Organ Culture Techniques; D011374:Progesterone / Q000494:pharmacology*; D011388:Prolactin / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis*; D011863:Radioimmunoassay
chemical_list: D002364:Caseins; D007328:Insulin; D012333:RNA, Messenger; D011374:Progesterone; D004958:Estradiol; D011388:Prolactin; D013006:Growth Hormone
references.citation: Biochemistry. 1974 Aug 13;13(17):3606-15
references.pmid: 4858491
references.citation: Int Rev Cytol. 1976;47:1-97
references.pmid: 186428
references.citation: Annu Rev Biochem. 1976;45:721-46
references.pmid: 183602
references.citation: Nature. 1973 Nov 16;246(5429):159-60
references.pmid: 4586108
references.citation: Cell. 1976 Aug;8(4):557-72
references.pmid: 182385
references.citation: J Natl Cancer Inst. 1974 Nov;53(5):1387-93
references.pmid: 4473562
references.citation: Recent Prog Horm Res. 1970;26:287-308
references.pmid: 4919093
references.citation: Recent Prog Horm Res. 1975;31:175-211
references.pmid: 1105717
references.citation: Endocrinology. 1976 Aug;99(2):445-51
references.pmid: 954644
references.citation: J Immunol Methods. 1975 Dec;9(2):123-34
references.pmid: 1206224
references.citation: Biochemistry. 1975 Jul;14(13):2895-903
references.pmid: 1148182
references.citation: J Endocrinol. 1975 Apr;65(1):1-6
references.pmid: 1141803
references.citation: Cell Differ. 1975 May;4(2):113-22
references.pmid: 1169123
references.citation: Biochemistry. 1975 Jan 14;14(1):69-78
references.pmid: 1109593
references.citation: J Biol Chem. 1969 Aug 25;244(16):4406-12
references.pmid: 5806584
references.citation: J Endocrinol. 1966 Aug;35(4):331-40
references.pmid: 5923706
references.citation: Endocrinology. 1961 Aug;69:340-53
references.pmid: 13741937
references.citation: J Natl Cancer Inst. 1959 May;22(5):1003-17
references.pmid: 13655076
references.citation: J Biol Chem. 1951 Nov;193(1):265-75
references.pmid: 14907713
references.citation: Recent Prog Horm Res. 1973;29:417-55
references.pmid: 4356276