help button home button Endocrine Society Endocrinology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Crandall, D. L.
Right arrow Articles by Kral, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Crandall, D. L.
Right arrow Articles by Kral, J. G.
Right arrowPubmed/NCBI databases
*Substance via MeSH
Endocrinology Vol. 140, No. 1 154-158
Copyright © 1999 by The Endocrine Society

ARTICLES

Angiotensin II Receptors in Human Preadipocytes: Role in Cell Cycle Regulation

David L. Crandall, Douglas C. Armellino, Dennis E. Busler, Barbara McHendry-Rinde and John G. Kral

Wyeth-Ayerst Research, Princeton, New Jersey 08543; and the Department of Surgery, State University of New York Health Science Center (J.G.K.), Brooklyn, New York 11203

Address all correspondence and requests for reprints to: Dr. David L. Crandall, Wyeth-Ayerst Research, CN 8000, Princeton, New Jersey 08543. E-mail: crandad{at}war.wyeth.com


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
The role of angiotensin II (AII) in human preadipocyte physiology has been investigated in primary cultures from human adipose tissue. Receptor binding studies indicated that human preadipocytes express a high affinity AII binding site of the AT1 subtype, as binding of 125I-labeled [Sar1,Ile8]AII was rapid, saturable, and specific. As AII has previously been demonstrated to affect the cell cycle in adrenal and cardiac cells, the effect of AII on regulation of cycle progression was examined in human preadipocytes. Stimulation of preadipocytes with AII resulted in G1 phase progression of the cell cycle, as determined by flow cytometric analysis. AII treatment was associated with induction of expression of the messenger RNA for the cell cycle regulatory protein cyclin D1 in a dose-dependent manner. Pretreatment of cells with subtype-selective AT receptor ligands before AII stimulation indicated that the cyclin response was mediated via the AT1 receptor. The identity of the cells as preadipocyte was verified by culture in a defined differentiation medium, observing both leptin message expression and triglyceride accumulation by flow cytometry. These findings indicate that AII has early, receptor-mediated effects on cell cycle progression in human preadipocytes that may contribute to differentiation to the adipocyte phenotype.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
ADIPOSE tissue is developmentally derived from a stem cell population that, in the preadipocyte stage, exhibits a fibroblast-like phenotype. During development, expression of key enzymes involved in lipogenesis transform the preadipocyte to the adipocyte phenotype, characterized by the storage of cytoplasmic lipid (1). The regulatory processes involved in the development of the adipocyte are important to a better understanding of the growth of adipose tissue in obesity and have most recently been examined by investigating the molecular trans-activators that become expressed during the stimulation of animal cells with hormones that classically have facilitated differentiation (1, 2). In murine preadipocyte cell lines, adipocyte differentiation also involves reentry into the cell cycle and a mitotic clonal expansion that is critical to the completion of the process of mature adipocyte development (1). This essential aspect of adipocyte differentiation has not been investigated in human preadipocytes, however.

Recent studies have suggested a role for angiotensin II (AII) in growth and development. Significant changes in AII receptor density with the stage of development of rat fetuses have been observed (3) as well as the expression of messenger RNA (mRNA) for AII receptors coinciding with cardiac development (4). Adipose tissue possesses the molecular machinery required for AII-stimulated signal transduction, including angiotensinogen (5, 6), high affinity membrane-bound AII receptors (7, 8), and angiotensin-converting enzyme (9). The function of the angiotensin system in adipose tissue remains under investigation, however, and has been primarily examined with respect to effects on mature adipocytes (10). A recent study of cell cycle progression in a human adrenal cell line (11), suggesting the involvement of AII in the expression of regulatory molecules of mitogenesis implies a novel role for AII that has not been examined in human adipose tissue. Using cells cultured from human adipose tissue, the present investigation has characterized the AII receptor in preadipocytes and then investigated a function for this receptor in cell cycle progression.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Culture of human adipocytes
Subcutaneous upper abdominal adipose tissue samples were obtained from men and women during elective operations. Written informed consent was obtained from all patients, and the protocol was approved by the institutional review board of the State University of New York Health Science Center at Brooklyn. A total of eight individual knife biopsies of adipose tissue were immediately placed in saline and transported to the laboratory. Using sterile technique, 10- to 15-g samples were cut into small pieces of approximately 100 mg each and placed in a sterile plastic Erlenmeyer flask containing 25 ml Krebs-Ringer bicarbonate buffer (pH 7.4), 6 mM glucose, and 2 mg/ml collagenase. The adipose tissue was shaken for 15 min at 100 strokes/min in a Dubnoff incubator maintained at 37 C. To facilitate the removal of stromal-vascular elements after the initial tissue digestion (8), the contents of the flask were passed through a sterile, 230-µm stainless steel tissue sieve (Cellector, Bellco Glass, Inc., Vineland, NJ) into a 50-ml sterile plastic test tube. The adipocytes were allowed to float to the top of the tube, the infranatant was collected and passed into another sterile tube, and the collagenase was neutralized with an equal volume of a solution containing medium 199, 10% heat-inactivated FCS, 100 U/ml penicillin, 100 µg/ml streptomycin, 250 ng/ml fungizone, and 2 mM L-glutamine (Life Technologies, Grand Island, NY), then spun immediately at 300 x g for 10 min. The supernatant from the centrifugation was discarded, and the pellet resuspended in 5 ml of the same medium containing serum, which was subsequently passed through a 25-µm steel sieve. The medium obtained from this final filtration was transferred to a sterile tissue culture flask (75 cm2) and maintained in an incubator at 37 C in 5% CO2. Cell attachment was allowed for 16–20 h, after which floating cells were removed by aspiration, followed by addition of fresh medium.

Receptor binding studies
Between passages 2–5, preadipocytes were trypsinized and plated in 24-well plates at an initial density of 25,000 cells/well. Cells were maintained in medium plus serum for 5–7 days after plating to allow attachment and proliferation. Proliferating preadipocytes were maintained in medium 199 supplemented with 10% heat-inactivated FCS and 1% antibiotic-antimycotic (Life Technologies). On the morning of the experiment, the medium was aspirated, and the cells were washed once with 1 ml PBS. The binding assay was begun by adding 200 µl binding buffer, 25 µl 1 µM unlabeled AII (nonspecific binding) or 25 µl buffer (total binding), and varying concentrations of 125I-labeled [Sar1,Ile8]AII (1100 Ci/mmol; 0.2–2.5 nM final concentration; Amersham, Arlington Heights, IL). The culture plates were shaken gently at room temperature for 90 min, and binding was terminated by aspiration of the medium. Each well was rinsed twice with 1 ml PBS, and preadipocytes were solubilized with 0.5 ml NaOH and counted for {gamma} radioactivity (Packard Cobra 5010 {gamma}-counter, 80% efficiency). For receptor subtype analysis, displacement of radioligand was determined in the presence of 1 x 10-5 to 1 x 10-10 M of the AT1 subtype-specific antagonist losartan or the AT2 subtype-specific ligand PD 123,319 [gifts from Dr. Ron Smith, DuPont-Merck Pharmaceutical Co. (Wilmington, DE) and Dr. Harvey Kaplan, Warner-Lambert (Ann Arbor, MI), respectively]. Cells were visualized microscopically throughout the experiment to qualitatively assess morphology, and representative wells were counted before and after the experiment using Coulter electronics (Hialeah, FL) to accurately determine cell number. Analysis of receptor number and affinity, and IC50 values for each subtype-specific ligand was performed using Lundon software (Lundon, Inc., Chagrin Falls, OH).

Flow cytometric analysis
Preadipocytes were plated at passage 1 or 2 in six-well plates and grown in medium 199, 10% FCS, and 1% antibiotic-antimycotic. At preconfluence, medium was removed and replaced with medium 199 containing 0.5% serum, and 24 h later fresh medium 199 with 0.5% serum with or without 1 µM AII was added in triplicate cultures. Human AII was obtained from Sigma Chemical Co. (St. Louis, MO) and solubilized in PBS on the morning of the experiment. After a 24-h period, untreated control and AII-stimulated preadipocytes were harvested and subjected to flow cytometric analysis. The relative percentage of cells in each stage of the cell cycle was analyzed using the Cycle TEST Plus DNA Reagent Kit (Becton Dickinson Co., San Jose, CA) according to the manufacturer’s specifications. The principle of this procedure is the labeling of nuclei with propidium iodide, followed by flow cytometric analysis (FACS Calibur, Becton Dickinson Co.).

Cyclin message determination
Preadipocytes used for determination of cyclin D1 expression were initially made quiescent by maintenance in medium containing 0.5% FCS for 24 h. After the initial 24-h period, AII was added, and total RNA (n = 5) was harvested from either control cells (0.5% FCS) or stimulated cells (0.5% FCS plus AII) following various protocols. RNA was extracted from control and AII-treated preadipocytes using the Qiagen RNeasy Mini Kit (Santa Clarita, CA) and was subjected to deoxyribonuclease digestion to eliminate genomic DNA. Total RNA was reverse transcribed using the SuperScript Preamplification System for First Strand cDNA Synthesis (Life Technologies) following the manufacturer’s instructions. Briefly, 1 µg total RNA was mixed with random hexamer primer and a 1-fold concentration of RT buffer containing 2.5 mM MgCl2. The reaction was incubated at 42 C for 50 min with 200 U SuperScript II reverse transcriptase (RT). The RT reaction was then stopped by incubation at 70 C, and residual RNA was digested in the presence of 2 U Escherichia coli ribonuclease H at 37 C for 20 min.

The complementary DNA (cDNA) was amplified by PCR by adding 2 µl of the RT reaction to 48 µl of a 1 x PCR Mastermix containing 12 pmol of each respective primer, 5 µl 10 x PCR buffer [200 mM Tris-HCl (pH 8.4), and 500 mM KCl], 4 µl 50 mM MgCl2, 1 µl deoxy (d)-NTP mixture (10 mM each of dATP, dCTP, and dTTP), and 2.5 U Taq polymerase. The cyclin D1 primers were designed to anneal to sequences from exons 2 and 5, spanning three introns, to give a 528-bp RT-PCR product (11). The sense primer sequence is 5'-GTC TGC GAG GAA GAG AAG-3', and the antisense sequence is 5'-GCA GGC CCG GAG GCA GTC-3'. The amplification protocol was for 30 cycles each at 94 C for 1 min, 56 C for 30 min, and 72 C for 1 min followed by an extension at 72 C for 5 min. A control reaction was concurrently performed by omitting RT from the reaction to confirm the absence of genomic contamination in cDNA samples. The human ribosomal 18S RNA was used as an internal standard and was amplified from the same cDNA using primers from the RT-PCR Quantum RNA system (Ambion, Inc., Austin, TX), yielding a 488-bp product. PCR products were separated on a 1.5% agarose gel, stained with syber green according to the manufacturer’s instructions (Molecular Probes, Inc., Eugene, OR), and visualized under short wave UV light. Imaging and quantitation were performed on a Storm 860 PhosphorImager (Molecular Dynamics, Inc., Sunnyvale, CA). No amplified products were detected in PCR reactions that lacked RT.

Preadipocyte differentiation
To verify that the population of cells being examined were, in fact, preadipocytes, each primary culture was stimulated to differentiate into the adipocyte phenotype by supplementing the standard growth medium with a medium containing 0.5 mM isobutylmethylxanthine, 0.1 µM hydrocortisone, and 10 pM insulin (12). Cultures were visualized daily by light microscopy during the differentiation period. After 1 week of differentiation, cytosolic triglyceride content was assessed by determining Nile Red uptake. Preadipocytes and adipocytes were incubated for 10 min at room temperature in a 10 µg/ml solution of Nile Red (Molecular Probes, Inc.), then subjected to flow cytometric analysis, with 10,000 events collected/sample. The 488 nm excitation light was used to generate forward scatter and right angle scatter signals. Nile Red fluorescence was detected at 530 nm, and data were expressed as a contour plot. In addition, RNA from control and differentiated cells was assessed for expression of leptin (13), with glyceraldehyde 3-phosphate dehydrogenase mRNA (14) expression determined from the same sample.

Statistical analysis was performed using Statistica/Mac (StatSoft, Tulsa, OK). The group mean ± SEM were considered significantly different at P < 0.05.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Preadipocytes were seeded in 24-well tissue culture plates, and AII receptor binding studies were performed on each of 8 separate cultures. Binding to cultured cells was rapid, saturable, and specific, and Scatchard analysis indicated a low capacity, high affinity receptor for AII (Fig. 1Go). The binding capacity (n = 8) was 15.5 ± 1.45 fmol/mg protein for pre-adipocytes, with a Kd of 1.5 nM ± 0.44 nM (±SEM). To identify the subtype of AII receptor, the IC50 for the AT1-specific inhibitor losartan and the AT2-specific ligand PD123,319 was determined in a range of concentrations from 1 x 10-5 to 1 x 10-10 M. Losartan inhibited binding in a dose-dependent manner with an IC50 of 2.12 x 10-8 M (n = 6; SEM = 1.07 x 10-8), whereas PD was ineffective in ligand displacement at the highest concentration tested (1 x 10-5 M), indicating human preadipocyte receptors are of the AT1 subtype.



View larger version (18K):
[in this window]
[in a new window]
  		Figure 1. Identification of AII receptors in human preadipocytes. Typical saturation binding isotherm of 125I-labeled [Sar1,Ile8]AII to cultured human sc preadipocytes. Each point represents the mean of specific binding in cells assayed in triplicate. Units of expression for bound ligand are femtomoles per mg protein x 101, whereas the free concentration is nanomolar. Scatchard transformation is shown in the inset.

 
The effect of AII on cell cycle progression as determined by flow cytometric analysis is shown in Table 1Go. The results of three typical, separate experiments from three individual cultures indicate that a 24-h treatment with 1 µM AII reduced the relative proportion of preadipocytes in G1 and increased the number of cells in the S and G2-M phases. Normalization of data in control cells between individuals allowed for comparison of trend analysis between treatments and indicated a significant reduction of cells in G1 phase and a significant increase in S and G2-M phases after AII treatment (P < 0.05).


View this table:
[in this window]
[in a new window]
  		Table 1. The percentage of cells within each phase of the cell cycle is shown for untreated, control preadipocytes and those treated for 24 h with 10-6 M AII

 
To determine whether AII had specific effects on expression of regulatory proteins involved in cell cycle progression, preadipocyte cyclin D1 mRNA was assessed by RT-PCR. As shown in Fig. 2Go, a 6-h treatment of preadipocytes with AII was associated with a significant increase in the expression of cyclin D1 mRNA in a dose-dependent manner. The effect of pretreatment of cells with selective AII receptor ligands followed by stimulation with AII and determination of cyclin D1 mRNA expression is shown in Fig. 3Go. The selective AT1 receptor antagonist losartan significantly attenuated the stimulatory response to AII, whereas the selective AT2 ligand PD 123,319 was ineffective. Statistical analysis indicated that AII significantly increased (P < 0.05) cyclin D1 mRNA expression. PD 123,319 values were similar to AII-stimulated values. Conversely, losartan-treated preadipocyte expression of cyclin D1 mRNA was similar to control values and significantly (P < 0.05) less than AII-stimulated values, indicating that the AII response was selectively inhibited by losartan. The AT1-mediated cyclin D1 response to AII therefore agrees with the ligand binding studies identifying the subtype expression of AII receptors in human preadipocytes as AT1.



View larger version (14K):
[in this window]
[in a new window]
  		Figure 2. Cyclin D1 mRNA expression in human preadipocytes after stimulation with AII. Quiescent preadipocyte cultures (n = 3) were stimulated with varying concentrations of AII, and RNA was harvested 6 h later and assessed for cyclin D1 expression by RT-PCR. Fold induction is shown with comparison to 18S mRNA in the same sample. *, Significantly different from the control value at P < 0.05.

 


View larger version (21K):
[in this window]
[in a new window]
  		Figure 3. Effect of angiotensin receptor subtype on AII-mediated cyclin D1 expression in human preadipocytes. a, Quiescent preadipocytes were treated for 1 h with 1 x 10-5 M of either the AT1-selective antagonist losartan or the AT2-selective ligand PD 123,319, followed by stimulation with 1 x 10-6 M AII. RNA was harvested from pre-adipocytes 6 h later, and the fold induction of cyclin D1 expression was determined by RT-PCR. Each treatment represents three to six separate experiments. *, Significantly different from the control value at P < 0.05). The value for losartan-treated cells is significantly different from AII-treated cells (P < 0.05). b, Representative gel from the RT-PCR experiments: lane 1, control; lane 2, AII-stimulated; lane 3, losartan-treated; lane 4, PD 123,319-treated.

 
Finally, to verify the primary cultures as being preadipocyte, each culture was maintained in a defined differentiation medium for 1 week, followed by flow cytometric analysis. During the differentiation period, preadipocytes maintained a fibroblast-like appearance, whereas the morphology of the differentiated cells changed to a rounded cell body with multiple lipid droplet inclusions. Figure 4Go shows the comparative flow cytometric analysis of the two culture treatments. The data are contour plotted according to both relative fluorescence, which is indicative of Nile Red uptake into cytosolic triglyceride, and side-scatter fluorescence, which is an indication of cellular morphology. Stimulation of the human preadipocytes to differentiate resulted in the appearance of an adipocyte population of cells that was not apparent in the preadipocyte cultures, characterized by an increase in Nile Red uptake in 100% of the cells analyzed and an increase in side-scatter fluorescence. In addition to triglyceride accumulation, differentiation was associated with expression of leptin mRNA, further verifying the cell population as human preadipocyte (Fig. 4Go).



View larger version (25K):
[in this window]
[in a new window]
  		Figure 4. Differentiation of human preadipocytes. a, Contour plot of side scatter (abscissa) vs. Nile Red uptake (ordinate) after 1 week of culture of human preadipocytes in a defined differentiation medium. The data are generated from flow cytometric analysis of cells incubated briefly with the fluorescent dye Nile Red. b, Leptin mRNA expression is preadipocytes and adipocytes subjected to identical culture conditions as those used for flow cytometric analysis. Glyceraldehyde 3-phosphate dehydrogenase mRNA is shown from the same sample.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Components of the renin-angiotensin system were first identified in peripheral tissues over a decade ago, with adipose tissue representing the potentially largest source of the precursor molecule, angiotensinogen (5, 6). As obesity and hypertension often coexist (15), synthesis of the pressor peptide hormone angiotensinogen by adipose tissue has been proposed as etiological (6, 16). In heart, brain, and kidney (4, 17, 18) angiotensin II has a role in morphological development. For this reason it is intriguing to consider a similar function for AII in the development of adipose tissue, one of the largest tissues in the body. Furthermore, data obtained from cultured murine adipocytes support a causal role for AII in development, as ligand binding to the AT2 receptor significantly increases the activities of two important lipogenic enzymes, fatty acid synthase and glycerol-3-phosphate dehydrogenase, and elevates cytosolic triglyceride content (10). Mechanistically, AII induces differentiation of murine preadipocytes through an AT2-dependent generation of prostacyclin (19), and microdialysis experiments in rat adipose tissue have substantiated the AII-stimulated release of prostacyclin in vivo (20).

Earlier studies from our laboratory investigated the role of the AII receptor in preadipocyte proliferation, determining that although human cells up-regulate AT1 receptor expression after differentiation, long term culture with exogenous AII has no effect on preadipocyte proliferation (21). The absence of a proliferative response prompted examination of other roles for this receptor in adipose tissue physiology. The current study has identified a high affinity, subtype-selective AII receptor in human preadipocytes and has examined the role of this receptor in cell cycle progression. The results indicate that the human preadipocyte receptor is AT1 subtype specific, in contrast to published studies identifying the AT2 subtype in human brain and heart (22, 23) as well as in murine adipocytes (10). In a logical progression of the novel discovery of this receptor in the human preadipocyte, experiments were needed to provide insight on receptor function. Our hypothesis that AII may regulate the preadipocyte cell cycle was based upon separate reports of the effects of AII on cell cycle progression in adrenal cortical cells (11) and the fact that 3T3-L1 cells must undergo a mitotic clonal expansion that is essential to terminal differentiation to the adipocyte phenotype (24). Regulatory molecules involved in this critical stage of adipocyte differentiation are just beginning to be investigated, among which are included the retinoblastoma and cyclin proteins (24). Cyclin D1 is a regulatory subunit of the G1 phase cyclin kinases, it is required for G1 phase progression, and its expression is necessary for proliferation (25). Our observations of expression of cyclin D1 mRNA as early as 6 h after AII treatment together with the progression of preadipocytes from the G1 phase suggest a novel role of AII in adipose tissue development. As clonal expansion of 3T3-L1 preadipocytes precedes permanent withdrawal from the cell cycle, leading eventually to terminal differentiation (26), these data suggest that AII may influence the earlier stages of development of preadipocytes as well as promoting lipogenic enzyme expression in mature adipocytes (10).

These studies identifying a new function of the adipocyte AII receptor in the regulation of cell cycle progression made extensive use of flow cytometric analysis of human primary cultures of preadipocytes. Those few investigations that have examined cell cycle progression and differentiation of pre-adipocytes have done so exclusively in murine cells, where cell cycle analysis and differentiation were assessed by bromodeoxyuridine labeling and staining with a lipophilic dye, respectively (24, 26). The techniques of flow cytometry we employed significantly expand the analysis of both cell cycle progression and adipocyte differentiation. With respect to cell cycle, we were able to accurately determine the relative proportion of cells in each stage of cycle progression, whereas bromodeoxyuridine staining simply provides a measure of DNA synthesis. The use of a fluorescent dye in the analysis of differentiation followed by flow cytometric analysis provided information on individual cellular morphology and relative amount of triglyceride in each cell. From our data it is obvious that not all human cells respond identically when stimulated to differentiate. Future experiments comparing the molecular components of a differentiated human adipocyte to those of one that is resistant to this phenotypic challenge may provide strategies for modifying cell recruitment and growth with the goal of preventing obesity. Although further investigations are required to better understand the physiological role of the AII receptor, the adipocyte is a cell that is readily sampled from normal weight and obese subjects and may be central to the etiology of cardiovascular disease and diabetes, the leading causes of death and disability in industrialized nations.

Received April 2, 1998.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. MacDonald OA, Lane MD 1995 Transcriptional regulation of gene expression during adipocyte differentiation. Annu Rev Biochem 64:345–373[CrossRef][Medline]
  2. Amri EZ, Ailhaud G, Grimaldi P 1991 Regulation of adipose tissue cell differentiation. II. Kinetics of induction of the aP2 gene by fatty acids and modulation by dexamethasone. J Lipid Res 32:1457–1463[Abstract]
  3. Millan MA, Carvallo P, Izumi SI, Zemel S, Catt K, Aguilera G 1989 Novel sites of expression of functional angiotensin II receptors in the late gestation fetus. Science 244:1340–1342[Abstract/Free Full Text]
  4. Everett AD, Fisher A, Tufro-McReddie A, Harris M 1997 Developmental regulation of angiotensin type 1 and 2 receptor gene expression and heart growth. J Mol Cell Cardiol 29:141–148[CrossRef][Medline]
  5. Harp JB, DiGirolamo M 1995 Components of the renin-angiotensin system in adipose tissue: changes with maturation and adipose mass enlargement. J Gerontol 50A:B270–B276
  6. Frederich RC, Kahn BB, Peach MJ, Flier JS 1992 Tissue-specific nutritional regulation of angiotensinogen in adipose tissue. Hypertension 19:339–344[Abstract/Free Full Text]
  7. Crandall DL, Herzlinger HE, Saunders BD, Zolotor RC, Feliciano L, Cervoni P 1993 Identification and characterization of angiotensin II receptors in rat epididymal adipocyte membranes. Metabolism 42:511–515[CrossRef][Medline]
  8. Crandall DL, Herzlinger HE, Saunders BD, Armellino DC, Kral JG 1994 Distribution of angiotensin II receptors in rat and human adipocytes. J Lipid Res 35:1378–1385[Abstract]
  9. Crandall DL, Herzlinger HE, Cervoni P, Scalea TM, Kral JG 1991 Vasoactive substances in human adipose tissue biopsies. In: Ailhaud G, Guy-Grand B, LaFontan M, Riquier D (eds) Obesity in Europe. Libbey, London, pp 405–409
  10. Jones BH, Standridge MK, Moustaid N 1997 Angiotensin II increases lipogenesis in 3T3–L1 and human adipose cells. Endocrinology 138:1512–1519[Abstract/Free Full Text]
  11. Watanabe G, Lee RJ, Albanese C, Rainey WE, Batlle D, Pestell RG 1996 Angiotensin II activation of cyclin D1-dependent kinase activity. J Biol Chem 271:22570–22577[Abstract/Free Full Text]
  12. Hauner H, Entenmann G, Wabitsch M, Gaillard D, Ailhaud G, Negrel R, Pfeiffer EF 1989 Promoting effect of glucocorticoids on the differentiation of human adipocyte precursor cells cultured in a chemically defined medium. J Clin Invest 84:1663–1670
  13. Vidal H, Auboeuf D, de Vos P, Staels B, Riou JP, Auwerx J, Laville M 1996 The expression of ob gene is not acutely regulated by insulin and fasting in human abdominal subcutaneous adipose tissue. J Clin Invest 98:251–255[Medline]
  14. Allen R, Trach K, Hoch J 1987 Identification of the 37-kDa protein displaying a variable interaction with the erythroid cell membrane as glyceraldehyde 3-phosphate dehydrogenase. J Biol Chem 262:649–653[Abstract/Free Full Text]
  15. Larsson B, Svardsudd K, Welin L, Wilhelmsen L, Bjorntorp P, Tibblin G 1984 Abdominal adipose tissue distribution, obesity, and risk of cardiovascular disease and death: 13 year follow-up of participants in the study of men born in 1913. Br Med J 288:1401–1404
  16. Cooper R, McFarlane-Anderson N, Bennett F, Wilks R, Puras A, Tewksbury D, Ward R, Forrester T 1997 ACE, angiotensinogen and obesity: a potential pathway leading to hypertension. J Hum Hypertension 11:107–111[CrossRef][Medline]
  17. Tsutsumi K, Saavedra JM 1991 Characterization and development of angiotensin II receptor subtypes (AT1 and AT2) in rat brain. Am J Physiol 261:R209–R216
  18. Tufro-McReddie A, Johns DW, Geary KM, Dagli H, Everett AD, Chevalier RL, Carey RM, Gomez RA 1994 Angiotensin II type 1 receptor: role in renal growth and gene expression during normal development. Am J Physiol 266:F911–F918
  19. Safonova I, Aubert J, Negrel R, Ailhaud G 1997 Regulation by fatty acids of angiotensinogen gene expression in preadipose cells. Biochem J 322:235–239
  20. Darimont C, Vassaux G, Gaillard D, Ailhaud G, Negrel R 1994 In situ microdialysis of prostaglandins in adipose tissue: stimulation of prostacyclin release by angiotensin II. Int J Obesity 18:783–788
  21. Crandall DL, Armellino DA, Kral JG 1997 Human adipocyte AII receptors in primary culture: role in proliferation and differentiation. Obesity Res [Suppl] 5:50S
  22. Ge J, Barnes NM 1996 Alterations in angiotensin AT1 and AT2 receptor subtype levels in brain regions from patients with neurodegenerative disorders. Eur J Pharmacol 297:299–306[CrossRef][Medline]
  23. Brink M, Erne P, de Gasparo M, Rogg H, Schmid A, Stula P, Bullock G 1996 Localization of the angiotensin II receptor subtypes in the human atrium. J Mol Cell Cardiol 28:1789–1799[CrossRef][Medline]
  24. Richon VM, Lyle RE, McGehee RE 1997 Regulation and expression of retinoblastoma proteins p107 and p130 during 3T3–L1 adipocyte differentiation. J Biol Chem 272:10117–10124[Abstract/Free Full Text]
  25. Hunter T, Pines J 1994 Cyclins and cancer: cyclin D and CDK inhibitors come of age. Cell 79:573–582[CrossRef][Medline]
  26. Shao D, Lazar MA 1997 Peroxisome proliferator activated receptor {gamma}, CCAAT/enhancer-binding protein {alpha}, and cell cyle status regulate the commitment to adipocyte differentiation. J Biol Chem 272:21473–21478[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 EndocrinologyHome page
L. Yvan-Charvet, F. Massiera, N. Lamande, G. Ailhaud, M. Teboul, N. Moustaid-Moussa, J.-M. Gasc, and A. Quignard-Boulange
Deficiency of Angiotensin Type 2 Receptor Rescues Obesity But Not Hypertension Induced by Overexpression of Angiotensinogen in Adipose Tissue
Endocrinology, March 1, 2009; 150(3): 1421 - 1428.
[Abstract] [Full Text] [PDF]

Home page
 J EndocrinolHome page
B Galvez-Prieto, J Bolbrinker, P Stucchi, A I de las Heras, B Merino, S Arribas, M Ruiz-Gayo, M Huber, M Wehland, R Kreutz, et al.
Comparative expression analysis of the renin-angiotensin system components between white and brown perivascular adipose tissue
J. Endocrinol., April 1, 2008; 197(1): 55 - 64.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Clin. Nutr.Home page
J. Koska, N. Stefan, P. A Permana, C. Weyer, M. Sonoda, C. Bogardus, S. R Smith, D. R Joanisse, T. Funahashi, J. Krakoff, et al.
Increased fat accumulation in liver may link insulin resistance with subcutaneous abdominal adipocyte enlargement, visceral adiposity, and hypoadiponectinemia in obese individuals
Am. J. Clinical Nutrition, February 1, 2008; 87(2): 295 - 302.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
V. Achard, S. Boullu-Ciocca, R. Desbriere, G. Nguyen, and M. Grino
Renin receptor expression in human adipose tissue
Am J Physiol Regulatory Integrative Comp Physiol, January 1, 2007; 292(1): R274 - R282.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. Rev.Home page
M. Paul, A. Poyan Mehr, and R. Kreutz
Physiology of local Renin-Angiotensin systems.
Physiol Rev, July 1, 2006; 86(3): 747 - 803.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. GenomicsHome page
A. S. Heimann, M. H. Favarato, F. C. Gozzo, V. Rioli, F. R. Carreno, M. N. Eberlin, E. S. Ferro, J. H. Krege, and J. E. Krieger
ACE gene titration in mice uncovers a new mechanism for ACE on the control of body weight
Physiol Genomics, January 20, 2005; 20(2): 173 - 182.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Endocrinol. Metab.Home page
P. A. Permana, S. Nair, Y.-H. Lee, G. Luczy-Bachman, B. Vozarova de Courten, and P. A. Tataranni
Subcutaneous abdominal preadipocyte differentiation in vitro inversely correlates with central obesity
Am J Physiol Endocrinol Metab, June 1, 2004; 286(6): E958 - E962.
[Abstract] [Full Text] [PDF]

Home page
 DiabetesHome page
J. Janke, S. Engeli, K. Gorzelniak, F. C. Luft, and A. M. Sharma
Mature Adipocytes Inhibit In Vitro Differentiation of Human Preadipocytes via Angiotensin Type 1 Receptors
Diabetes, June 1, 2002; 51(6): 1699 - 1707.
[Abstract] [Full Text] [PDF]

Home page
 J. Nutr.Home page
S. Kim, J. Whelan, K. Claycombe, D. B. Reath, and N. Moustaid-Moussa
Angiotensin II Increases Leptin Secretion by 3T3-L1 and Human Adipocytes via a Prostaglandin-Independent Mechanism
J. Nutr., June 1, 2002; 132(6): 1135 - 1140.
[Abstract] [Full Text] [PDF]

Home page
 DiabetesHome page
E. Maquoi, C. Munaut, A. Colige, D. Collen, and H. R. Lijnen
Modulation of Adipose Tissue Expression of Murine Matrix Metalloproteinases and Their Tissue Inhibitors With Obesity
Diabetes, April 1, 2002; 51(4): 1093 - 1101.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
T. J. Smith, L. Koumas, A. Gagnon, A. Bell, G. D. Sempowski, R. P. Phipps, and A. Sorisky
Orbital Fibroblast Heterogeneity May Determine the Clinical Presentation of Thyroid-Associated Ophthalmopathy
J. Clin. Endocrinol. Metab., January 1, 2002; 87(1): 385 - 392.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
F. Massiera, J. Seydoux, A. Geloen, A. Quignard-Boulange, S. Turban, P. Saint-Marc, A. Fukamizu, R. Negrel, G. Ailhaud, and M. Teboul
Angiotensinogen-Deficient Mice Exhibit Impairment of Diet-Induced Weight Gain with Alteration in Adipose Tissue Development and Increased Locomotor Activity
Endocrinology, December 1, 2001; 142(12): 5220 - 5225.
[Abstract] [Full Text] [PDF]

Home page
 HypertensionHome page
T. Skurk, Y.-M. Lee, and H. Hauner
Angiotensin II and Its Metabolites Stimulate PAI-1 Protein Release From Human Adipocytes in Primary Culture
Hypertension, May 1, 2001; 37(5): 1336 - 1340.
[Abstract] [Full Text] [PDF]

Home page
 Journal of Renin-Angiotensin-Aldosterone SystemHome page
A. M Sharma and S. Engeli
The renin-angiotensin system in obesity hypertension
Journal of Renin-Angiotensin-Aldosterone System, March 1, 2001; 2(1_suppl): S114 - S119.
[PDF]

Home page
 EndocrinologyHome page
C. Ling, G. Hellgren, M. Gebre-Medhin, K. Dillner, H. Wennbo, B. Carlsson, and H. Billig
Prolactin (PRL) Receptor Gene Expression in Mouse Adipose Tissue: Increases during Lactation and in PRL-Transgenic Mice
Endocrinology, October 1, 2000; 141(10): 3564 - 3572.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
D. L. Crandall, D. E. Busler, B. McHendry-Rinde, T. M. Groeling, and J. G. Kral
Autocrine Regulation of Human Preadipocyte Migration by Plasminogen Activator Inhibitor-1
J. Clin. Endocrinol. Metab., July 1, 2000; 85(7): 2609 - 2614.
[Abstract] [Full Text]

Home page
 HypertensionHome page
S. Engeli, R. Negrel, and A. M. Sharma
Physiology and Pathophysiology of the Adipose Tissue Renin-Angiotensin System
Hypertension, June 1, 2000; 35(6): 1270 - 1277.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Crandall, D. L.
Right arrow Articles by Kral, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Crandall, D. L.
Right arrow Articles by Kral, J. G.
Right arrowPubmed/NCBI databases
*Substance via MeSH

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Endocrinology Endocrine Reviews J. Clin. End. & Metab.
Molecular Endocrinology Recent Prog. Horm. Res. All Endocrine Journals