1989 Fiscal Year Final Research Report Summary
Molecular mechanism of growth and differentiation of mammary glands.
| Project/Area Number |
63304024
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| Research Category |
Grant-in-Aid for Co-operative Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
畜産学(含草地学)
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| Research Institution | University of Tokyo |
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Principal Investigator |
KOHMOTO K. Univ. Tokyo, Fac.Agric., Professor, 農学部, 教授 (30011894)
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| Co-Investigator(Kenkyū-buntansha) |
ENAMI J. Dokkyo Med. College, Lecturer, 講師 (30112634)
SAKAI S. Univ. Tokyo, Fac. Agric. Assoc. Prof., 農学部, 助教授 (80114487)
NAGASAWA H. Meiji Univ., Fac. Agric. Professor, 農学部, 教授 (10076987)
YOKOYAMA A. Nagoya Univ., Fac. Agric. Professor, 農学部, 教授 (50023388)
KOMINE S. Kyorin Univ., Sch. Med., Professor, 医学部, 教授 (40086484)
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| Project Period (FY) |
1988 – 1989
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| Keywords | cell proliferation / cell culture / growth factor / pregnancy-dependent mammary tumor / mouse / Suncus murinus / prolactin receptor / inositol-1,4,5-triphosphate |
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| Research Abstract |
1. Establishment of mammary cell culture systems and effects of growth factors: I)To investigate the interaction between hormones and growth factors on the mammary epitherium, we established a serum-free cell culture system of mammary cells which includes the culture of mammary cells embedded in cool agen gels and culture with physiological concentrations of insulin, IGF-I, EGF, prolactin, progesterone and vitamin D_3. Insulin, IGF-I and EGF independently stimulated growth of cells whereas other hormones stimulated growth only with insulin or the growth factor. These results suggest that these hormones can stimulate growth only by modulating the action of growth factors. (2) The incorporation of deoxybromuridine into mammary cells of Suncus murinus revealed that oestradiol stimulates the growth of ducts and that progesterone forms lateral buds. (3)A conditioned medium factor derived from fibroblast cultures was purified and its antibody was raised.
2. Pregnancy-dependent mouse mammary t
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umor and normal growth of mammary glands (1)Pregnancy-dependent mammary tumor is a hormone-dependent mammary tumor and gives a good model for the investigation of hormone action in normal mammary growth. Mice with this tumor-show poor lactational performance. Insight into the details reveals that the normal mammary cells of the mice functioned normally but that the mammary growth was depressed. Hormone administrations did not improve the performance, but tumorectomy did very much. This tumor may produce a factor suppressing growth of normal mammary growth. (2)Ten cell lines derived from pregnancy-dependent mammary tumor were established. There was no EGF autocrine cell line among these.
3. Mode of action of polypeptide hormones : (1)The endoplasmic reticulum and golgi apparatus were separated by ultracentrifugation from lactating mouse mammary glands. The release of Ca^<2+> from these fractions was confirmed by addition of inositol-1,4,5-triphosphate. The increase in phosphatidyl inositol-specific phospholipase C activities was also observed in organ-cultured mammary glands with insulin. This suggests the involvement of the inositol-1,4,5-triphosphate - Ca^<2+> release system in the signal transduction of hormone in mammary glands. (2) Prolactin receptor was purified from rabbit mammary, glands with ion exchange, con-A column and affinity chromatography. The thermodynamic kinetics of prolactin binding was also analyzed. Less
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