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1.
Development and characterization of SV40 immortalized rat submandibular acinar cell lines 总被引:3,自引:0,他引:3
D. O. Quissell K. A. Barzen D. C. Gruenert R. S. Redman J. M. Camden J. T. Turner 《In vitro cellular & developmental biology. Animal》1997,33(3):164-173
Summary Rat submandibular salivary gland acinar cells were transfected by CaPO4 precipitation using a plasmid containing a replication-defective simian virus (SV40) genome. Out of 27 clonal cell lines,
two were shown to have moderate to high levels of cytodifferentiation and salivary gland acinar cell function. Functional
studies with the two cell lines indicated that the β-adrenergic agonist, isoproterenol, vasoactive intestinal peptide, and
prostaglandin E1 were effective activators of intracellular cyclic AMP production. Epinephrine, norepinephrine, phenylephrine, acetylcholine,
and P2U-purinoceptor agonists were effective in increasing inositol phosphate production and intracellular free calcium levels, whereas
substance P was without effect. Utilizing indirect immunofluorescence analysis, both cell lines were shown to express glutamine/glutamic
acid-rich proteins, a submandibular acinar cell specific secretory protein family. Electron microscopic evaluation documented
the maintenance of tripartite junctional complexes, cellular polarization, and the presence of moderate amounts of secretory
granules and rough endoplasmic reticulum. The two cell lines had doubling times of 25 h. 相似文献
2.
Effects of dexamethasone,epidermal growth factor,and retinoic acid on rat submandibular acinar-intercalated duct complexes in primary culture 总被引:6,自引:0,他引:6
Robert S. Redman David O. Quissell Katherine A. Barzen 《In vitro cellular & developmental biology. Plant》1988,24(8):734-742
Summary Mature acini with attached segments of intercalated ducts were dissociated from the submandibular glands of rats and grown
in primary culture on gels of reconstituted rat tail collagen. Screening evaluations indicated that the following new conditions
promoted a substantial improvement in the survival of the cells as compared with our previously reported results: a) adding
dexamethasone, epidermal growth factor, and retinoic acid to the medium, b) decreasing the fetal bovine serum in the medium
to 1%; and c) adjusting the oxygen in the gas phase to 50%. A detailed evaluation, including light and electron microscopy
and biochemical analysis, then provided the following observations. The acinar-ductal complexes enlarged throughout the 22-d
culture period, and after 4d sheets comprised of a one- to two-cell thick layer of acinar cells spread among the complexes.
Synthesis of mucin, and its secretion in response to norepinephrine or cAMP, dropped precipitously to very low levels after
2 d. However, synthesis of DNA, general proteins, and glycoproteins dropped only transiently after 2 d, rising to levels approaching
those of freshly dissociated complexes by 22 d. These data indicate that a shift occurred from the synthesis of large quantities
of secretory proteins and glycoproteins, especially mucins, during the first 2d in culture, to other materials thereafter.
Overall, the new culture conditions resulted in substantial growth and survival of acinar cells through 22 d in primary culture,
but the important acinar characteristic of the synthesis and secretion of mucins was essentially lost after 4 d.
This investigation was supported by grant DK 33835 from the National Institutes of Health, Bethesda, MD, and the Medical Research
Service of the Veterans Administration, Washington, DC. 相似文献
3.
Short-term primary culture of acinar-intercalated duct complexes from rat submandibular glands 总被引:7,自引:0,他引:7
David O. Quissell Robert S. Redman Marvin R. Mark 《In vitro cellular & developmental biology. Plant》1986,22(8):469-480
Summary Acinar-intercalated duct complexes dissociated from rat submandibular glands have been shown to be an excellent model for
studying secretory responses of salivary gland components. However, they are functionally normal for only a few hours. We
undertook a systematic manipulation of primary culture conditions in an attempt to extend the useful life of the complexes.
The major modifications that were tested were increased oxygenation in increments to 95%; substitution of norepinephrine or
carbamylcholine or both for isoproterenol in the medium; different sources of collagen for and addition of laminin, fibronectin
and/or type IV collagen to the matrix gel; and varying the thickness of the collagen gel, richness of the cell suspension
inoculate, and sources and concentrations of sera in the medium. Progress was monitored by light microscopic evaluation of
routine sections of specimens until improved maintenance of acinar and other cells warranted carrying parallel cultures for
biochemical, histochemical, and ultrastructural analyses. Best results were obtained with 90% O2, laminin in rat tail collagen gel, 10% fetal bovine serum, and 3 μM isoproterenol. Morphologically, there was good survival of acini and intercalated ducts after 1 d, with decreased acinar
size being correlated with secretory response to the isoproterenol. Reorganization and considerable mitotic activity were
seen at 2, 3, and 4 d, with most clusters of cells becoming much larger than the original complexes. During this period acinar
cells steadily became less differentiated and their numbers decreased in proportion to intercalated duct or undifferentiated
cells. However, there was good overall survival through 7 d. Biochemical analysis indicated that the cells were able to maintain
significant biosynthetic activity for 4 d, with DNA, RNA, protein, and glycoprotein synthetic rates increasing over the culture
period, but the secretory capacity of the cells diminished during the primary culture period, with mucin biosynthesis and
secretion decreasing significantly after 1 d in culture.
This work was supported by a grant from the Thrasher Research Fund, by Grant AM 33835 from the National Institutes of Health,
Bethesda MD, and by the Medical Research Service of the Veterans Administration, Washington, D.C. 相似文献
4.
Limesand KH Barzen KA Sanders LA Sclafani RA Raynolds MV Reyland ME Anderson SM Quissell DO 《In vitro cellular & developmental biology. Animal》2003,39(3-4):170-177
Summary Apoptosis is a highly organized cellular process that is critical for maintaining glandular homeostasis. We have used primary
rat salivary acinar cells from the parotid and submandibular glands to investigate the critical regulatory events involved
in apoptosis. Caspase-3 activity, cleavage of caspase substrates, and deoxyribonucleic acid (DNA) fragmentation were assayed
in cells treated with etoposide, a DNA-damaging agent, or brefeldin A (BFA), a Golgi toxin. Dose-response studies showed that
the sensitivity of both cell types to etoposide and BFA was similar, with 150 μM etoposide or 1.5 μM BFA inducing maximal caspace activation. However, BFA induced a more robust activation of caspase and DNA fragmentation in
both cell types. Similar results were observed when the caspase cleavage of poly(adenosine 5′-diphosphate ribose) polymerase
and protein kinase C delta were analyzed by Western blot. Analysis of the kinetics of apoptosis showed that caspace-3 activation
was maximal at 8 h of etoposide or BFA treatment in the parotid cells and at 8–18 h in the submandibular cells. A similar
time course was observed when DNA fragmentation was assayed, although maximal DNA fragmentation in BFA-treated cells was two-to
threefold higher than that observed in etoposide-treated cells. Despite slight kinetic differences, it would appear that the
apoptotic cascade is very similar in both primary parotid and submandibular acinar cells. Although limited in their long-term
stability in culture, the use of primary, nonimmortalized salivary acinar cultures will also permit the use of specific transgenic
animals to further characterize the molecular events involved in the regulation of salivary gland acinar cell apoptosis. 相似文献
5.
Growth in primary culture of mouse submandibular epithelial cells embedded in collagen gels 总被引:8,自引:0,他引:8
Jason Yang Darragh Flynn Lisa Larson Susan Hamamoto 《In vitro cellular & developmental biology. Plant》1982,18(5):435-442
Summary Mouse submandibular glands were dissociated and the epithelial cells embedded in a collagen gel matrix. A characteristic and
reproducible pattern of growth was seen resulting in three-dimensional outgrowths with ductlike structures projecting into
the matrix. A sustained cell growth leading to a 5 to 10-fold increase in cell number was observed in less than 2 wk. The
extent of this growth was found to be dependent on serum concentration. Of the three sera tested, swine serum was found to
promote greater growth compared to fetal bovine serum or horse serum. Swine serum dose response studies have shown that a
concentration of 2 to 5% in the medium elicited only a modest increase, if any, in cell number compared to the initial value
within a period of 2 wk. Various hormones and growth factors were then added to this “maintenance” medium. Insulin was found
to stimulate growth consistently and reproducibly in a dose-dependent manner. Ultrastructurally, the resulting outgrowths
were comprised of polarized cells joined by apical tight junctions and desmosomes. These outgrowths produced epidermal growth
factor in response to dihydrotestosterone, triiodothyronine, and cortisol. The present system provides a method for sustaining
growth and functional differentiation in primary culture of mouse submandibular gland epithelial cells.
This investigation was supported by PHS Grants CA05388 and CA09041, awarded by the National Cancer Institute, Department of
Health and Human Services. 相似文献
6.
Miho Furue Tetsuji Okamoto Hiroyuki Hayashi J. Denry Sato Makoto Asashima Shigeru Saito 《In vitro cellular & developmental biology. Animal》1999,35(3):131-135
Summary To study the mechanisms of morphogenesis in salivary gland regeneration, we have established the RSMG-1 cell line derived
from submandibular gland (SMG) of 10-wk-old Wistar female rats in serum-free culture. Our finding that RSMG-1 cells originated
from duct cells was based on morphology and immunohistochemical results. In three-dimensional serum-free collagen gel culture,
HGF induced branching morphogenesis of RSMG-1 cells. Histological examination revealed that HGF-induced branching structure
exhibited well-formed lumina. This morphology closely resembles that found in vivo. The cells also expressed activin A. Exogenously
added activin A at a high concentration reduced HGF-induced branching morphogenesis. These findings suggest that the morphogenesis
of the salivary gland is modulated by HGF and activin A. Our results show that the RSMG-1 cell line may be useful in studies
of salivary gland regeneration. 相似文献
7.
Lawrence E. Shapiro Neil Wagner 《In vitro cellular & developmental biology. Plant》1988,24(4):299-303
Summary Serum-free tissue culture medium consisting of a 1∶1 mixture of Dulbecco's modified Eagle's medium (DMEM) and Ham's F12 medium
is herein shown to support growth of Reuber H-35 cells over several days in culture. Cells were initially plated in serum
containing DMEM medium for 3 h. After cell attachment, serum is removed and replaced with a serum-free 1∶1 mixture of these
two commercially available tissue culture media. The doubling time of cell growth in this unsupplemented serum-free medium
was 46 h in lightly plated cultures over the first 5 d. The presence of transferrin (5 μg/ml) and insulin (3.3 nM) results in a cell doubling time of 17 h, which equaled the growth rate in medium containing 10% fetal bovine serum. In the
absence of transferrin, growth rates in serum-free medium were correlated with the cell density of cultures. Conditioned medium
from dense, serum-free cultures has growth-stimulating activity in recipient lightly plated cultures. This simple, serum-free
culture medium will facilitate studies on the growth regulation of H-35 rat hepatoma cells.
This work was funded by a feasibility grant from the American Diabetes Association, as well as by the National Institutes
of Health grants CA 24604-09 and CA 16463-14. 相似文献
8.
M. P. Lopez M. J. Gomez-Lechon J. V. Castell 《In vitro cellular & developmental biology. Plant》1984,20(12):923-931
Summary Liver parenchymal cells cultured in serum-free medium may retain their ability to synthesize glycogen in response to insulin.
Specific hormone requirements are needed by hepatocytesto retain the biochemical pattern of mature cells. Insulin supplementation
of culture medium seems to be essential to maintain the glycogen synthesis rate of cultured hepatocytes. The continuous presence
of dexamethasone amplified the insulin-induced glucogen synthesis. Cytophotometric analysis showed differences in the way
that individual cells accumulate glycogen in response to insulin stimulus, which indicates that liver parenchymal cells in
culture are functionally heterogeneous.
The financial support for this work was from the Fondo de Investigationes Sanitarias de la Seguridad Social, grants 41/82
and 48/82. 相似文献
9.
Growth of mouse vaginal epithelial cells in culture: Effect of sera and supplemented serum-free media 总被引:3,自引:0,他引:3
Taisen Iguchi Francis-Dean A. Uchima Howard A. Bern 《In vitro cellular & developmental biology. Plant》1987,23(8):535-540
Summary Normal mouse vaginal epithelial cells isolated from ovariectomized ca. 35-d-old BALB/cCrgl mice were grown in primary culture
using collagen gel metrix and a serum-free medium composed of a 1∶1 mixture of Dulbecco’s modified Eagle’s medium and Ham’s
F12 (D:H) medium supplemented with insulin (IN), epidermal growth factor (EGF), cholera toxin, transferrin, and bovine serum
albumin V (BSA). Three-dimensional cellular outgrowths occurred inside the collagen gel matrix. The contribution of each factor
to cell growth was examined by individual addition to the basic D:H medium and by individual deletion from the complete serum-free
medium. When added individually, only IN promoted growth. Deletion of IN from the complete serum-free medium markedly, diminished
growth; deletion of EGF or BSA slightly diminished growth. When horse, fetal bovine, or chicken serum was added to the basal
D:H medium, only with increasing doses of horse serum was there enhanced cell growth. The effect of 17?-estradiol and diethylstilbestrol
on the growth of cells was also tested, using a suboptimal medium of D:H supplemented with BSA and IN, or a minimal medium
supplemented with IN alone. During the 8-d time period, addition of estrogen did not enhance cell growth in either medium.
To date, we have been unable to demonstrate a mitogenic effect of estrogen; rather a dose-dependent inhibition of proliferation
is seen.
This investigation was supported by grants CA-05388 and CA-09041, awarded by the National Institutes of Health, DHHS, Bethesda,
MD. 相似文献
10.
Insulin is often included in serum-free media for animal cell cultivation. However, the necessity of insulin for a specific cell line is rather uncertain. In this article we report the effects of insulin on the cultivation of a hybridoma cell line in a serum-free medium. It was found that insulin affected neither the cell growth nor the antibody production. The specific growth rate and specific antibody production rate were very similar in the cultures with or without insulin. However, the presence of insulin affected the nutrient consumption rate and cell metabolism. Including insulin in the medium resulted in a higher specific glucose consumption rate, a shorter exponential growth stage, and a lower final antibody concentration. The elimination of insulin from the medium allowed antibody to accumulate to a concentration substantially higher than that in the insulin-containing cultuvre. (c) 1995 John Wiley & Sons, Inc. 相似文献
11.
A cloned rat thymic epithelial cell line established from serum-free selective culture 总被引:6,自引:0,他引:6
Arthur Piltch Paul Naylor Jun Hayashi 《In vitro cellular & developmental biology. Plant》1988,24(4):289-293
Summary A serum-free system has been developed for selective growth and long-term culture of rat thymic epithelial cells. The growth
media is a modification of McKeehan's WAJC 404, plus insulin, cholera toxin, dexamethasone, and epidermal growth factor. Cultures
have been continuously passaged and maintained for over 6 mo., and a cloned cell line, TEA3A1, has been established. These
cells are epithelial, judging by morphology and ultrastructure, and are positive for A2B5 and thymosin α markers for thymic
endocrine cells.
This work was partly supported by grant PCM-834 0582 from the National Science Foundation, Washington, DC, and grant P01 CA
37589-2 from the National Cancer Institute, Bethesda, MD. 相似文献
12.
David G. Thomassen 《In vitro cellular & developmental biology. Plant》1989,25(11):1046-1050
Summary The colony-forming efficiency of rat tracheal epithelial (RTE) cells was determined in serum-free media containing different
types of commercially available bovine serum albumin (BSA): crude fraction V, essentially globulin-free, essentially fatty-acid-free,
and essentially globulin- and fatty-acid-free BSA. RTE cells exhibited a concentration-dependent increase in colony-forming
efficiency in response to crude fraction V BSA. Similar results were obtained using essentially globulin-free BSA. However,
deletion of cholera toxin from the medium resulted in a decrease in the colony-forming efficiency for cells plated in high
concentrations (>2 mg/ml) of globulin-free, but not one type of fraction V, BSA. Essentially fatty-acid-free or essentially
fatty-acid- and globulin-free BSA stimulated RTE cell colony formation at low concentrations (less than 2.5 to 5 mg BSA/ml)
but resulted in concentration-dependent decreases in colony-forming efficiency at higher concentrations. The response of cells
to these BSAs was not dependent on cholera toxin. Finally, commerically available fraction V BSA prepared by heat shock, dialysis,
charcoal treatment, and deionization was stimulatory at low concentrations but inhibitory at high concentrations. These data
suggest that impure preparations of BSA can, under different conditions, stimulate or inhibit cell proliferation and that
the expression, of these activities is affected by the method of BSA preparation, the concentration of BSA used, and, in some
cases, by the presence or absence of cholera toxin.
Research conducted with support from the Office of Health and Environmental Research, U.S. Department of Energy, Washington,
DC, under contract no. DE-AC04-76EV01013 in facilities fully acredited by the American Association for Laboratory Animal Care. 相似文献
13.
Label-retaining cells in the rat submandibular gland. 总被引:1,自引:0,他引:1
Masaya Kimoto Yoshiaki Yura Mitsunobu Kishino Satoru Toyosawa Yuzo Ogawa 《The journal of histochemistry and cytochemistry》2008,56(1):15-24
To identify stem cells in salivary glands, label-retaining cells (LRCs) were established in rat submandibular glands. Developing and regenerating glands were labeled with bromodeoxyuridine (BrdU). To cause gland regeneration, the glands were injured by duct obstruction. BrdU LRCs were observed in all the parenchymal structures except for the acinus of the glands labeled during regeneration. Among these LRCs, a few, but not many, expressed neither keratin18 (K18; an acinar/duct cell marker) nor alpha-smooth muscle actin (alphaSMA; a myoepithelial cell marker), and thus were putative stem cells. These (K18 and alphaSMA)(neg) LRCs were invariably observed in the intercalated duct and the excretory duct. In the intercalated duct, they were at the proximal end bordering the acinus (the neck of the intercalated duct). Next, to test the above identification, gland extirpation experiments were performed. LRCs were established by labeling developing glands with iododeoxyuridine (IdU) in place of BrdU. Removal of one submandibular gland forced the IdU-LRCs in the remaining gland to divide. They were labeled with chlorodeoxyuridine (CldU). The (K18 and alphaSMA)(neg) LRCs in the neck of the intercalated duct and in the excretory duct did not change in number or in IdU label. The CldU label appeared in these cells and then disappeared. These results indicate that the (K18 and alphaSMA)(neg) LRCs have divided asymmetrically and are thus considered salivary gland stem cells. 相似文献
14.
LONGTMR3IGF-I, an analogue of insulin-like growth factor (IGF)-I, was specifically engineered for use in biopharmaceutical protein
production in mammalian cells. LONGTMR3IGF-I is capable of supporting the growth and survival of Chinese hamster ovary cells in serum-free media at concentrations
at least 200-fold lower than required for insulin. LONGTMR3IGF-I also acts as a more potent growth and survival factor than either insulin or native IGF-I in SF culture of human embryonic
kidney (HEK293) cells. To investigate the basis of the enhanced potency of LONGTMR3IGF-I we have examined the mechanism of action of these mitogens in HEK293 cells. All mitogens tested were found to activate
the TypeI IGF receptor (IGF-IR) and insulin receptor (IR) in a dose-responsive manner. However, the level of activation of
both receptors after stimulation with LONGTMR3IGF-I, at lower concentrations, was greater than with either insulin or IGF-I. The greater potency of LONGTMR3IGF-I in activating the IR, despite having a low affinity for IRs, suggests the presence of heterotetrameric IGF-IR/IR dimers.
Interestingly, the decrease in IGF-IR activation at higher concentrations of LONGTMR3IGF-I suggests that the dose-response curve may be bell-shaped. 相似文献
15.
Di Lorenzo Teresa P. De Maro Joseph A. Pumo Dorothy E. 《In vitro cellular & developmental biology. Plant》1989,25(10):909-913
Summary A serum-free culture system was used to compare the nutritional requirements of mouse mammary cells transformed by bovine
papillomavirus type 1 (ID13 cells) and the uninfected parent line (C127 cells). The serum-free, chemically defined medium
used for this study was an MCDB 151-based medium (MCDB 151+S+I), supplemented with epidermal growth factor, transferrin, hydrocortisone,
ethanolamine, phosphoethanolamine, retinoic acid, trace metals, and insulin. Proliferation of either cell type in serum-free
culture required the addition of 250 μg/ml of insulin. ID13 cells have a doubling time of greater than 96 h in MCDB 151+S+I,
whereas C127 cells have a doubling time of 60 h. This is in sharp contrast to the growth characteristics of the two cell types
in 10% fetal bovine serum, where doubling times for the ID13 and C127 cells are 24 and 30 h, respectively. Culture of the
cells in a serum-free medium has therefore revealed that the papillomavirus-transformed cells have more stringent growth requirements
than the uninfected parent line.
This work was supported in part by grant #1-P01 NS19214 from the National Institutes of Health, Bethesda, MD, NSF grant #R11-8217798
from the National Science Foundation, Washington, DC, and by a grant from the Otolaryngology Foundation. 相似文献
16.
DAS M Bhargava N Gregory C Riedel L Molnar P Hickman JJ 《In vitro cellular & developmental biology. Animal》2005,41(10):343-348
Summary In this study, we have documented by morphological analysis, immunocytochemistry, and electrophysiology, the development of
a culture system that promotes the growth and long-term survival of dissociated adult rat spinal cord neurons. This system
comprises a patternable, nonbiological, cell growth-promoting organosilane substrate coated on a glass surface and an empirically
derived novel serum-free medium, supplemented with specific growth factors (acidic fibroblast growth factor, heparin sulfate,
neurotrophin-3, brain-derived neurotrophic factor, glial-derived neurotrophic factor, cardiotrophin-1, and vitronectin). Neurons
were characterized by immunoreactivity for neurofilament 150, neuron-specific enolase, Islet-1 antibodies, electrophysiology,
and the cultures were maintained for 4–6 wk. This culture system could be a useful tool for the study of adult mammalian spinal
neurons in a functional in vitro system. 相似文献
17.
Willie D. Morgan Jeannette E. Williams Cecil W. Lee Clyde J. Dawe 《In vitro cellular & developmental biology. Plant》1979,15(12):1013-1022
Summary Time-lapse phase-contrast cinematography revealed contractile activity within mouse submandibular salivary gland rudiments
in organotypic culture. Three types of contraction were distinguishable. In type I (voiding contractions), all portions of
the gland contracted synchronously, and the active state ranged from 30 min to 2 hr. In type II (priming contractions), all
portions of the gland contracted synchronously, but the active state was shorter, ranging from 4 to 10 min. In type III (churning
contractions), isolated foci in lobules or secretory units throughout the gland contracted asynchronously and had very short
active states of about 1 min. By electron microscopy, myoepithelial cells could first be demonstrated in submandibular glands
developing either in vitro or in vivo, at 21 days postconception. Contractions in the cultured rudiments began as early as
18 days postconception. Since neither smooth nor striated muscle could be identified in these glands by electron microscopy,
the contractions are believed to result from myoepithelial activity that apparently may begin before ultrastructural evidence
of myoepithelial differentiation is contractile function and indirect evidence has lent ample support to this presumption,
the present study represents the first direct cinematographic demonstration and characterization of myoepithelial contractions,
under conditions in vitro. 相似文献
18.
Development of embryonic chick insulin cells in culture: Beneficial effects of serum-free medium, raised nutrients, and biomatrix 总被引:1,自引:0,他引:1
Summary A previous finding that insulin cells do not survive or differentiate in explants of embryonic avian pancreas cultured in
collagen gel with a serum-containing medium has provided a model system for identification of conditions favorable for development
of these cells. To this end, we here modify the substrate and the medium. The epithelial component of dorsal pancreatic buds
of 5-d chick embryos was cultured for 7 d on Matrigel in serum-containing and in serum-free medium, the latter incorporating
insulin, transferrin, and selenium, Endocrine cell types were distinguished by immunocytochemistry; insulin cell counts were
expressed as a proportion of insulin plus glucagon cells. With serum-containing medium, Matrigel stimulated a significant
increase in this proportion as compared with collagen gel—3.1% as against 0.2%; the serum-free medium further increased this
proportion to 17.3%. Raising the level of essential amino acids approximately fivefold increased the latter figure somewhat
(to 18.9%), but it was more than doubled (to 37.4%) by raising the glucose concentration from 10 mM to 20 mM. Raising the levels of amino acids and glucose simultaneously yielded a lesser increase (to 31.8%). Some cultures grown in
collagen gel and serum-containing medium for 7 d were transferred to Matrigel and serum-free medium for a further 7 d. Insulin
cell development recovered, indicating that progenitor cells had survived and were stimulated to develop by the improved conditions.
This study indicates that components of the biomatrix and the medium (in particular, a raised glucose concentration) are important
for the survival and differentiation of embryonic insulin cells. 相似文献
19.
Ostuni MA Tumilasci OR Péranzi G Cardoso EM Contreras LN Arregger AL Papadopoulos V Lacapere JJ 《Biology of the cell / under the auspices of the European Cell Biology Organization》2008,100(7):427-439
Background information. TSPO (translocator protein), previously known as PBR (peripheral‐type benzodiazepine receptor), is a ubiquitous 18 kDa transmembrane protein that participates in diverse cell functions. High‐affinity TSPO ligands are best known for their ability to stimulate cholesterol transport in organs synthesizing steroids and bile salts, although they modulate other physiological functions, including cell proliferation, apoptosis and calcium‐dependent transepithelial ion secretion. In present study, we investigated the localization and function of TSPO in salivary glands. Results. Immunohistochemical analysis of TSPO in rat salivary glands revealed that TSPO and its endogenous ligand, DBI (diazepam‐binding inhibitor), were present in duct and mucous acinar cells. TSPO was localized to the mitochondria of these cells, whereas DBI was cytosolic. As expected, mitochondrial membrane preparations, which were enriched in TSPO, exhibited a high affinity for the TSPO drug ligand, 3H‐labelled PK 11195, as shown by Bmax and Kd values of 10.0±0.5 pmol/mg and 4.0±1.0 nM respectively. Intravenous perfusion of PK 11195 increased the salivary flow rate that was induced by muscarinic and α‐adrenergic agonists, whereas it had no effect when administered alone. Addition of PK 11195 also increased the K+, Na+, Cl− and protein content of saliva, indicating that this ligand modulated secretion by acini and duct cells. Conclusions. High‐affinity ligand binding to mitochondrial TSPO modulates neurotransmitter‐induced salivary secretion by duct and mucous acinar cells of rat submandibular glands. 相似文献
20.
Masatoshi Togami Daphne Blazka Jun Hayashi 《In vitro cellular & developmental biology. Plant》1988,24(7):699-704
Summary A serum-free, hormone and attachment factor supplemented culture for rat H4 hepatoma cells was established. In the defined
medium (Dulbecco's Modified Eagle's +Ham's F12+insulin, transferrin, fibronectin liver cell growth factor, and sodium selenite),
H4 cells grew equally well as in 10% fetal bovine serum supplemented medium. H4 cells in either defined or serum-containing
culture conditions produce transferrin but not albumin or alpha-fetoprotein. In this paper we have studied the effect of various
hormones and pressor peptides on the production of angiotensinogen by H4 cells cultured in defined conditions. Only glucocorticoid
hormone had a significant effect on the production of angiotensinogen, whereas other hormones previously reported to exert
their effect on angiotensinogen production had little or no effect.
This work was supported by grant P01 CA37589 from the National Institutes of Health, Bethesda, MD. 相似文献