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1.
Growth of mouse vaginal epithelial cells in vitro 总被引:1,自引:0,他引:1
B. A. Flaxman D. P. Chopra D. Newman 《In vitro cellular & developmental biology. Plant》1973,9(3):194-201
Summary Pieces of adult mouse vagina (comprising epithelium and connective tissue), when explanted onto glass coverslips, gave rise
to outgrowing sheets of pure epithelium whose cells had ultrastructural features in common with the cells of origin in vivo.
Epithelial outgrowths from vaginas of estradiol-primed and nonprimed ovariectomized mice were studied. After the first 5 days
in vitro, in the absence of estradiol, the labeling index and length of the cell cycle were similar in both types of cultures.
The values were similar to those reported by others in vivo in response to estrogen. Thus, proliferative activity of cells
from nonprimed mice was stimulated merely by in vitro conditions, while proliferation of cells from primed mice continued
at the high level existing prior to explantation. The high rate of proliferation wasnot associated with keratinization of any cells. In the continued absence of estrogen, cells from both kinds of cultures showed
a progressive decrease in proliferative activity between 5 and 14 days, also associated with inability of cells to keratinize.
Addition of estradiol didnot reverse the mitotic drop or promote keratinization. Supplementation with hydrocortisone and insulin had no effect. The results
suggest that (a) vaginal epithelial cells in vitro require factors in addition to estradiol in order to maintain a high level of proliferative
activity or to differentiate fully by keratinization and (b) keratinization is not dependent on rate of cell proliferation.
Supported by grants from the National Cancer Institute (1 PO 1 CA 11536) and the National Institute of Arthritis and Metabolic
Diseases (1 P0 1 AM 15515). 相似文献
2.
Jill M. Siegfried Karen G. Nelson Jane L. Martin David G. Kaufman 《In vitro cellular & developmental biology. Plant》1984,20(1):25-32
Summary Histochemical techniques have been applied to the identification of cell types cultured from human endometrium. Previous work
from this laboratory characterized two principtal cell types found in cultures of endometrium: a mature epithelial cell and
another cell which was classified as the endometrial stromal cell based on light and electron microscopy. In this report we
compare the histochemical staining of endometrial tissue in frozen sections to that of cultured cells. These results confirm
the epithelial and stromal nature of the respective cell types. Several markers were found that could distinguish between
cells of epithelial and stromal origin. The enzymes alkaline phosphatase, γ-glutamyltranspeptidase, peroxidase, and β-glucuronidase
were localized in glandular and surface epithelia in frozen sections and in colonies of epithelial cells in culture. Stroma
in frozen sections and cultured stromal cells contained leucine aminopeptidase and fibronectin. Epithelial sections and in
culture could also be distinguished from cells of stromal origin by preferential binding of lotus and peanut lectin. Several
other markers were found in both endometrial epithelium and stroma.
J. M. S. was recipient of National Research Service Award CA09156 (National Cancer Institute); K. G. N. was recipient of National
Research Service Award ES07017 (National Institute of Environmental Health Sciences); and D. G. K. was recipient of Research
Career Development Award CA00431 from the National Cancer Institute, Bethesda, MD. Supported by Grant CA 31733 from the National
Cancer Institute, Bethesda, MD. 相似文献
3.
Paul S. Cooke Dennis K. Fujii Gerald R. Cunha 《In vitro cellular & developmental biology. Plant》1987,23(3):159-166
Summary Vaginal and uterine stromal (VS and UtS) cells have been cultured in a collagen gel matrix, and the ability of the cells to
retain their identity and interact normally with epithelia after culture was examined. Stromal explant from 2-d-old mice were
plated onto an extracellular matrix covered with collagen, and maintained in Ham’s F12∶DMEM (1∶1) containing 15% fetal bovine
serum. The fibroblastic stromal cells invaded and eventually filled the overlying collagen during the 4-wk growth period,
and the total DNA of the UtS and VS cultures increased 3.5- and 4-fold, respectively. To assess the ability of the cultured
stroma to perform its normal functions after the in vitro period, recombinations of cultured stroma and fresh epithelia were
preparaed and transplanted under the renal capsule of female hosts and grown for 4 wk. The epithelium in recombinants of cultured
VS + vaginal epithelium (VE) and cultured UtS + uterine epithelium (UtE) was histologically normal and proliferated in response
to estrogen. Cultured stroma also instructively induced heterologous epithelium; VS induced UtE to undergo vaginal differentiation,
and UtS induced VE to undergo uterine differentiation. These results indicate that UtS and VS retain their identity and do
not irreversibly dedifferentiate in culture. Stromal cells grown in a colagen gel matrix form a functional stroma; they interact
normally with epithelium after culture and express normal permissive and instructive inductive functions when reassociated
with epithelium and grown in vivo.
This work was supported by grants HD 17491, AM/CA 16570, CA 05388, and 5 F32 HD06580 from the National Institute of Health,
Bethesda, MD, and a grant from the UCSF Academic Senate. 相似文献
4.
Summary The effect of variations in culture conditions on survival of fragments of mouse and rat descending colon in organ culture
was studied by morphological and functional criteria. A combination of conditions demonstrated to be beneficial permitted
maintenance for at least 35 days. These included: a gaseous environment of 95% O2:5% CO2, an attachment matrix consisting of a Metricel GA-4 membrane (pore size, 0.8 μ), intermittent exposure to the gas and fluid
phases by rocking in 5 ml medium and supplementation of the medium with 1.0 μM dexamethasone and 10% FBS. During this time,
the crypt structure of the mucosal epithelium was well preserved, and DNA synthesis in the crypts and mucin production in
the crypts and superficial epithelium continued. In addition, the synthetic hormone, pentagastrin, stimulated DNA synthesis
in the mucosal epithelium of mouse colon fragment in short-term organ culture.
This work was supported by Environmental Protection Agency Grant R803998-01-1 and National Cancer Institute Contract N01-CP-75952. 相似文献
5.
Joseph L. Melnick Janet S. Butel Satvir S. Tevethia Nilambar Biswal Matilda Benyesh-Melnick 《In vitro cellular & developmental biology. Plant》1971,6(5):349-354
Summary This paper describes some current work pertaining to transformation of cells by oncogenic viruses.
Part I includes: (1) the effect of a deoxyribonucleic acid (DNA) tumor virus (SV40) on the antigenic characteristics of transformed
cells; (2) in vitro and in vivo methods of detecting virus-specific surface antigens; (3) the role that the host cell may
play in the expression of virus-coded antigens; and (4) the presence of virus-induced antigens as a possible mechanism of
the apparent nononcogenicity of certain virus variants.
Part II discusses (1) the physicochemical properties of the nucleic acid of a ribonucleic acid (RNA) tumor virus-the Moloney
sarcoma-leukemia virus (MSV-MLV) complex —(2) a preliminary analysis of viral RNA replication in cells transformed by MSV-MLV,
and (3) application to human tumors.
Supported in part by Research Grant CA 04600 and by Research Contract PH 43-68-678 within the Special Virus-Cancer Program,
National Cancer Institute, National Institutes of Health.
Recipient of Research Career Development Award 5-K3-CA 38,614 from the National Cancer Institute, National Institutes of Health 相似文献
6.
B. Allen Flaxman 《In vitro cellular & developmental biology. Plant》1972,8(3):237-250
Summary The normal steady state in human epidermis reflects a balance between the rates of cell proliferation and of cell differentiation
(keratinization). In certain diseases, such as psoriasis and basal cell cancer, increased proliferative activity is associated
with abnormalities in keratinization. These events have been thought to be directly related. However, studies in which the
epithelial cells from normal epidermis, psoriasis, and basal cell cancer were grown in vitro suggest that abnormalities of
the keratinization process do not directluy result from hyperproliferation. Alterations in keratinization probably result
from abnormal dermal-epidermal interactions, independent of any dermal effects on mitosis.
This work was supported by grants from the National Cancer Institute (1 PO 1 CA 11536) and the National Institute of Arthritis
and Metabolic Disease (1 PO 1 AM 1551-01). 相似文献
7.
Richard I. Schwarz Deborah A. Farson Mina J. Bissell 《In vitro cellular & developmental biology. Plant》1979,15(12):941-948
Summary Primary avian tendon cells (PAT) maintain their embryonic state when cultured in medium F-12 with very low serum (0.2%) and
ascorbate (50 μg per ml); that is, they retain the potential for devoting 20–30% of their total protein synthesis to collagen.
However, if the cells are left at a confluent cell density or are derived from confluent cultures, this potential is irreversibly
decreased. This effect, along with poor medium formulations, probably accounts for the “dedifferentiation” process that occurs
when fibroblasts are cultured. In contrast, PAT cells kept at subconfluent cell densities retain the ability to synthesize
high levels of collagen. The one limitation in obtaining long-term cultures of high collagen-producing tendon cells in the
inability of serum at low concentrations to remain a potent mitogen after a few subcultures.
The quantitative loss of function has long been considered to be a cell culture artifact; however, we propose that this drop
in collagen synthesis is a reflection of the developmental programing of these cells. In separate series of experiments using
organ cultures, we show that tendon tissue from the embryo makes over 30% collagen, whereas, “young” tendons make 18% and
“older” tendons from the adult make less than 1%. Therefore, a quantitative drop in collagen synthesis would be expected if
normal development were to occur in culture. Our data are consistent with the idea that cultures of embryonic tendon cells
are triggered to mature by a mechanism that correlates with high cell density.
This investigation was supported in part by National Science Foundation Grant PCM 77-14982; in part by the Division of Biomedical
and Environmental Research of the Department of Energy under contract W-7405-ENG-48; and by a National Institutes of Health
Fellowship IF32 CA 05807-01, from the National Cancer Institute to R. I. S. 相似文献
8.
Cell identification in primary cell cultures from skin 总被引:2,自引:0,他引:2
B. Allen Flaxman 《In vitro cellular & developmental biology. Plant》1974,10(1-2):112-118
Summary Primary cell cultures can readily be obtained from human skin using the explant method. With special care it is possible to
obtain primary cultures consisting of epidermal keratinocytes without fibroblast contamination. By means of differences in
their growth patterns and retention of specific differentiative functions in vitro, keratinocytes and fibroblasts can easily
be distinguished. The high degree of confidence in establishing cell identity permits meaningful experimental use of this
system. The method of enzymatic separation of epidermis from dermis, followed by culture of cells from the dissociated epidermal
tissue, provides both epithelial and dendritic cells. The former are probably keratinocytes, whereas the latter are definitely
melanocytes. The possibility of eventual fibroblast overgrowth, using this latter method, has not yet been ruled out with
certainty.
Presented at the Workshop on Primary Cell Culture, November 1–3, 1973, Convenor Dr. Warren I. Schaeffer, University of Vermont,
at the W. Alton Jones Cell Science Center, Lake Placid, N. Y. The editorial assistance of Dr. and Mrs. Joseph Leighton in
preparing for press the five papers from that workshop is gratefully acknowledged.
This work was supported by grants from the National Cancer Institute, 1 PO 1 CA 11536, and the National Institute of Arthritis
and Metabolic Diseases, 1 PO 1 AM 15515. 相似文献
9.
Michael Solursh Patricia Buckley Ahrens Rebecca S. Reiter 《In vitro cellular & developmental biology. Plant》1978,14(1):51-61
Summary Tissue-culture methods can be used to test the developmental capacity of embryonic cells. In micro-mass cultures, derived
from wing cells of stages 21 through 24 chick embryos, aggregates of cells form and then differentiate into cartilage nodules,
as judged by the presence of an Alcian blue staining extracellular matrix. Wing cells derived from embryos as young as stage
17 can form aggregates. However, unless they are treated with db cyclic AMP and theophylline, it is not until stage 20 that
these aggregates can produce cartilage in culture. In clonal cell culture, cartilage colonies are not produced by primary
cell suspensions of limb cells until stage 25 when overt cartilage differentiation is occurring in vivo. It is possible to
obtain clonable cartilage cells from limb cells from embryos between stages 20 and 24 if the cells are either treated with
db cyclic AMP and theophylline or maintained in suspension culture for 12 to 48 hr. On the basis of these in vitro results
a multiple step model for the conversion of limb mesenchyme into cartilage cells is proposed. The model involves the appearance
of cells with a predisposition to form aggregates, development of the capacity to form cartilage in response to elevated levels
of cyclic AMP, the appearance of receptors that translate changes in either cell shape or cell cycle parameters into elevated
levels of cyclic AMP, aggregation, elevated levels of cyclic AMP, cartilage cell determination, and differentiation. This
model can serve as the basis for further tests.
Presented in the Opening Symposium on Nutritional Factors and Differentiation at the 28th Annual Meeting of the Tissue Culture
Association, New Orleans, Louisiana, June 6–9, 1977.
This work was supported by USPHS Training Grant HD00152 from the National Institute of Child Health and Human Development,
while P.B.A. was a postdoctoral trainee, and by NIH Grant HD05505 to M.S. 相似文献
10.
A method for the continuous cultivation of mammary epithelium 总被引:1,自引:0,他引:1
Etienne Y. Lasfargues Dan H. Moore 《In vitro cellular & developmental biology. Plant》1971,7(1):21-25
Summary Established cell lines of mammary epithelium have been obtained from mice, rats, and hamsters. Maintenance and replication
of the epithelium in serial subcultures were dependent on their periodic treatment with collagenase. Because collagenase is
not cytotoxic and has maximum efficiency at neutral pH in isotonic saline solutions containing calcium and magnesium, this
enzyme can be introduced directly into the culture medium; cells have been maintained for 3 days in such a medium with serous
enrichment at no detriment to them. Up to 10% concentrations of serum have not interfered with enzymatic activity.
Supported by United States Public Health Service Grant CA-08515 and CA-08740 from the National Cancer Institute. General Research
Support Grant FR-5582 from the National Institutes of Health, and Grant-in-Aid Contract M-43 from the State of New Jersey. 相似文献
11.
Boyd R. Switzer George K. Summer 《In vitro cellular & developmental biology. Plant》1973,9(3):160-166
Summary In human diploid skin fibroblasts in culture we have shown that nonhydroxylated collagen precursors remain in the cell when
proline hydroxylation is inhibited by α, α′-dipyridyl, a chelator of ferrous ions. The inhibition of proline hydroxylation
is reversed by addition of fresh medium containing 50 μg per ml of sodium ascorbate, whereupon nonhydroxylated collagen precursors
are hydroxylated within the cell and extruded into the medium. Extrusion of collagen already formed within the cell is not
appreciably affected by α, α′-dipyridyl inhibition. Under normal conditions collagen is released from the monolayer into the
medium within 3 hr of a pulse ofL[14C]proline. In the presence of α, α′-dipyridyl, about 35% of theL[14C]proline incorporated into protein is released into the medium within 8 hr as a proline-rich, hydroxyproline-deficient protein;
at the same time, approximately 15% of the protein-boundl-[14C]proline remains in the cell for as long as 12 hr. When proline hydroxylation is restored after 2 and 12 hr of α, α′-dipyridyl
inhibition, approximately the same amount of hydroxyproline is formed after each time interval in the monolayer. Therefore,
nonhydroxylated collagen precursors retained in the cell are not appreciably degraded during at least 12 hr of inhibition
by α, α′-dipyridyl and are extruded into the medium only upon restoration of hydroxylation.
This work was supported in part by a grant from the Easter Seal Research Foundation, and by Project 236, Health Services and
Mental Health Administration, Department of Health, Education and Welfare, Grant HD-03110 from the National Institute of Child
Health and Human Development, an American Cancer Society Institutional Grant (1N 15-J), a General Research Support Award (5-S01-FR-05406)
from the National Institutes of Health, a University Research Council Grant, a National Science Foundation Equipment Grant
(GB-4577), and a Research Career Development Award (5-K3-AM-5058) from the National Institute of Arthritis and Metabolic Disease
(G.K.S.). 相似文献
12.
L Sankaran P Qasba Y J Topper 《Biochemical and biophysical research communications》1984,125(2):682-689
Mammary tissue from rats that had been ovariectomized and adrenalectomized 4 weeks previously was compared to that from intact rats in terms of epithelial content and hormone-responsiveness in vitro. The endocrinectomy resulted in about a 30% enlargement of the gland, but led to a loss of only about 12% of the epithelium. This estrogen-depleted epithelium was able to acquire full responsiveness in vitro to insulin in terms of the accumulation of alpha-aminoisobutyric acid, and induction of glucose-6-phosphate and gluconate-6-phosphate dehydrogenases. It was also fully responsive to cortisol in relation to the induction of NADH-cytochrome C reductase, and to prolactin in terms of total RNA synthesis. However, estrogen-depletion led to an 82% loss in the ability of a unit amount of the epithelium to synthesize casein in response to these 3 hormones, and to a similar loss in relation to the accumulation of 25K casein mRNA. Estrogen administration in vivo could prevent and reverse the casein lesion. The disparity between constitutive and casein hormone-responsiveness in the absence of estrogen is discussed in relation to cell commitment. 相似文献
13.
Jose Russo Megan J. Milis Michelline J. Moussalli Irma H. Russo 《In vitro cellular & developmental biology. Plant》1989,25(7):643-649
Summary In experimental animal models the susceptibility of the mammary gland to neoplastic transformation is related to its degree
of development and proliferative activity; this observation led us to determine whether the human breast epithelium also exhibits
development-related differences, and whether these differences could be detected in an in vitro system. Normal breast tissue
obtained from reduction mammoplasties of 9 patients ranging in age from 18 to 56 years were characterized in both whole mount
preparations and organoids obtained after collagenase-hyaluronidase digestion by their degree of development based upon the
types of lobules present. Lobules were classified into type 1 (Lob 1), composed of approximately 11 alveolar buds, the less
developed; lobules type 2 (Lob 2), of moderate development, composed of approximately 47 ductules each, and lobules type 3
(Lob 3), composed of 80 ductules each, represented the highest level of development. Epithelial organoids obtained after digestion
were plated in DMEM:F12 medium supplemented with hydrocortisone, cholera toxin, insulin and 5% horse serum with a calcium
concentration of 1.05 mM Ca++. Following attachment, the medium was replaced by medium containing 0.040 mM Ca++. The percentage of attachment of organoids to the flask was greater in cells from Lob 1 (89–99%) and Lob 1+2 (79–100%) than
in cells from Lob 3, which had a 53–67% attachment. The total yield of cells after 7 weeks in culture was also greater in
cells derived from Lob 1 and Lob 1+2 than in cells from Lob 3. The total yield of cells obtained from primary cultures was
not related to the number of organoids plated, but to the degree of development of the gland. The DNA-labeling index (DNA-LI)
in intact breast tissue correlated with that in primary cultures; it was greater in Lob 1 and Lob 1+2 than in Lob 3. By flow
cytometry, the highest percentage of cells in S-phase was seen in cells with the highest DNA-LI. We concluded that the growth
characteristics of mammary epithelial cells in vitro in a low Ca++ medium is modulated by the degree of development and differentiation of the gland.
Supported by PHS Grant CA-38921 awarded by the National Cancer Institute, DHHS, and an Institutional Grant from the United
Foundation of Greater Detroit. 相似文献
14.
Fatemeh Fazely Dorothy C. Moses Nada Ledinko 《In vitro cellular & developmental biology. Plant》1985,21(7):409-414
Summary Invasion of chick chorioallantoic membrane (CAM) organ cultures by rat 3Y1 cells transformed by the highly oncogenic human
adenovirus type 12 (3Y1/12-10 cells) was inhibited by several retinoids tested. The anti-invasive activity of the retinoids
was dependent on retinoid concentration and continuous (4d) exposure of the CAM. The 50% retinoid dose (dose effective in
achieving a response in half of the organ cultures) that inhibited invasion was 0.85 μg/ml of retinol palmitate, 0.39 μg/ml
of retinoic acid, or 0.16 μg/ml of retinol acetate. This dose was of the same order of magnitude as that which induced CAM
differentiation, and was three-to fourfold less than the dose that caused cytotoxic damage of CAM. In addition, the retinoids
inhibited 3Y1/12-10 cell growth by approximately 40% at levels over 10-fold higher than those needed for anti-invasion activity.
The findings suggest that the anti-invasive activity of retinoids was at least partly due to direct induction of cell differentiation
of the CAM host tissue.
This work was supported by National Cancer Institute Grant CA 13231 and by University of Akron Grant RG 832. 相似文献
15.
Summary Using the Hayashi method, the distribution of glucosaminidase was studied throughout sequential molar development. Conspicuous activity was observed in certain cellular components of the stellate reticulum and dental papilla. The outer enamel epithelium, stratum intermedium and ameloblasts displayed less intense reactions.Supported by PHS Grant No. 2800-02 —Tooth Germ Development, National Institute of Dental Research, National Institutes of Health.Lieutenant Commander, Dental Corps, United States Navy. 相似文献
16.
Linda E. Malick Anna Tompa Charles Kuszynski Parviz Pour Robert Langenbach 《In vitro cellular & developmental biology. Plant》1981,17(11):947-955
Summary The maintenance of primary cultures of adult hamster pancreatic cells on layers of irradiated C3H/10T1/2 cells was studied.
Various types of pancreatic cells, acinar, islet and ductular cells could be identified in the cultures by light and electron
microscopy. Morphologically the various pancreatic cells retained many differentiated characteristics of their respective
in vivo cell types. Insulin production was maintained at near Day 1 levels for the 16 d in culture for which it was measured.
Colonies of epithelial cells continued to grow during a 20 d culture period. It is believed that this procedure for maintaining
functional and growing pancreas cells in culture may be a useful in vitro model for studying the initiation of pancreatic
carcinogenesis.
Supported by Grant R01 CA 20022 and Contract N01 CP33278 from the National Cancer Institute, National Institutes of Health,
Bethesda, Maryland. 相似文献
17.
Robert A. Harper B. Allen Flaxman 《In vitro cellular & developmental biology. Plant》1981,17(5):393-396
Summary Primary cell cultures of normal rabbit epidermal cells (keratinocytes) were established without the use of enzymatic techniques.
Six experiments were carried out on cells from six different rabbits. When these cells were exposed to methotrexate (MTX)
for 24 h at 1 μg/ml, proliferation, as measured by cells entering mitosis, was significantly inhibited (P<0.05) in only one experiment. When the dose of MTX was elevated to 100 μg/ml, only two experiments showed significant inhibition
of mitosis. This minimal inhibition of mitosis by MTX was contrasted by the dramatic inhibitory effect of this antimetabolite
on DNA synthesis. At 1 μg/ml MTX for 24 h, DNA synthesis, as measured by [3H]deoxyuridine uptake, was inhibited >95%. We can conclude that under certain conditions, the rabbit keratinocyte may represent
a normal cell type that is inherently resistant to the antiproliferative effects of methotrexate.
The research was supported by National Cancer Institute Grant CA 11536. 相似文献
18.
Summary Cells of embryos carrying a lethal nucleolar mutation have been maintained in vitro for extended periods of time. Normally
these mutants live only 9 to 12 days after fertilization but their cells in culture will survive for more than 3 months. The
extent of ribosomal RNA (rRNA) synthesis was determined in primary cultures prepared from normal embryos and nucleolar mutants
having different numbers of ribosomal RNA genes. We found that the accumulation of radioactivity into rRNA for normal and
mutant embryos was similar in vivo and in vitro. In primary cultures of normal embryos which have two nucleoli per cell and
mutant embryos which have only one nucleolus per cell, the incorporation of radio-activity into rRNA was similar even though
the normal cells have twice as many rRNA genes. Thus the mechanism which regulates dosage compensation of the rRNA genes operates
both in vivo and in vitro.
This work was supported by Grant GB38651 from the National Science Foundation. 相似文献
19.
Neil I. Goldstein Paul B. Fisher 《In vitro cellular & developmental biology. Plant》1978,14(2):200-206
Summary In the present study we tested the feasibility of utilizing a structurally modified polyene antibiotic, amphotericin B methyl
ester (AME), as a half-selection agent for isolating somatic cell hybrids. By using HAT medium supplemented with AME we have
isolated interspecific mouse-hamster hybrids from mixed cultures of mouse (TK−C1ID or HPRT− A9) and hamster (BHK/C 13) cells fused with Sendai virus, lysolecithin or polyethylene glycol. Hybrid cells proliferated
and clones were isolated after 2 to 3 weeks growth in three changes of HAT-AME medium and subsequent growth in HAT medium
alone. In contrast, genetically deficient parental C1 1D or A9 cells and AME-sensitive BHK/C 13 cells were killed using a
similar growth protocol. The described technique is simple, efficient and permits one to use a cell line without a genetic
defect in combination with a genetically deficient cell type in hybrid formation.
This investigation was supported in part by Contract NIH 69-2161, NIH Grant No. AI-2095 and NIH Training Grant No. GM 507
from the National Institute of General Medical Sciences. 相似文献
20.
The effects of sequential administration of 17 beta-estradiol on the synthesis and secretion of specific proteins in the immature rat uterus 总被引:1,自引:0,他引:1
We report here results of a study of the effect of sequential administration of 1 microgram 17 beta-estradiol in vivo on the incorporation of L-[35S]methionine into specific proteins in vitro in the immature rat uterus. One-dimensional SDS-PAGE analysis of labeled secreted uterine proteins and cellular proteins extracted from the luminal epithelial and from the stroma plus myometrial uterine fractions revealed that estradiol preferentially stimulated the synthesis of 110 K, 74 K and 66 K secreted proteins, 180 K and 110 K epithelial proteins and a 175 K stroma-myometrial protein among others, while it decreased the relative rate of synthesis of a 32.5 K secreted protein and a 70 K stroma-myometrial protein. The 110 K protein, a secreted luminal epithelial protein whose labeling in vitro dramatically increased greater than 60-fold per mg endometrial DNA after in vivo estrogen stimulation, may be a useful marker for studying estrogen action in the luminal epithelium of the immature rat uterus. Comparison of the secreted proteins labeled at 28 h (4 h after a second injection) and at 54 h (6 h after a third injection) revealed that estradiol effected a sequential change in the pattern of synthesis of secreted uterine proteins in vitro. Comparison of the number and magnitude of changes in the synthesis of specific proteins in the luminal epithelium and the stroma plus myometrium revealed that protein synthesis in the luminal epithelium is clearly more responsive to estradiol and readily distinguishable from the responsiveness of the stroma plus myometrium. 相似文献