- Volume 16 Issue 12
DOI QR Code
Molecular Cloning of Adipose Tissue-specific Genes by cDNA Microarray
- Kim, Kee-Hong (Department of Nutritional Sciences and Toxicology, University of California) ;
- Moon, Yang Soo (Department of Nutritional Sciences and Toxicology, University of California)
- Received : 2003.05.21
- Accepted : 2003.08.27
- Published : 2003.12.01
In an attempt to isolate novel molecules that may play a regulatory role in adipocyte differentiation, we devised an experimental strategy to identify adipose tissue-specific genes by modifying cDNA microarray technique. We used genefilter membranes containing approximately 15,000 rat non-redundant EST clones of which 4,000 EST were representative clones of known genes and 11,000 ESTs were uncharacterized clones. A series of hybridization of genefilter membranes with cDNA probes prepared from various rat tissues and nucleic acids sequence analysis allowed us to identify two adipose-tissue specific genes, adipocyte-specific secretory factor (ADSF) and H-rev107. Verification of tissue-specific expression patterns of these two genes by Northern blot analysis showed that ADSF mRNA is exclusive expressed in adipose tissue and the H-rev107 mRNA is predominantly expressed in adipose tissue. Further analysis of gene expression of ADSF and H-rev107 during 3T3-L1 adipocyte differentiation revealed that the ADSF and H-rev107 gene expression patterns are closely associated with the adipocyte differentiation program, indicating their possible role in the regulation of adipose tissue development. Overall, we demonstrated an application of modified cDNA microarray technique in molecular cloning, resulting in identification of two novel adipose tissue-specific genes. This technique will also be used as a useful tool in identifying novel genes expressed in a tissue-specific manner.
Supported by : Agricultural R&D Program Center of Ministry of Agriculture and Forestry
- Akiyama, H., Y. Hiraki, M. Noda, C. Shigeno, H. Ito and T. Nakamura. 1999. Molecular cloning and biological activity of a novel ha-ras suppressor gene predominantly expressed in skeletal muscle, heart, brain, and bone marrow by differential display using clonal mouse ec cells, ATDC5. J. Biol. Chem. 274: 32192-32197.
- Gerhold, D., T. Rushmore and C. T. Caskey. 1999. DNA chips: Promising toys have become powerful tools. Trends Biochem. Sci. 24:168-173.
- Hajnal, A., R. Klemenz and R. Schafer. 1994. Subtraction cloning of h-rev 107, a gene specifically expressed in h-ras resistant fibroblasts. Oncogene 9:479-490.
- Halaas, J. L., K. S. Gajiwala, M. Maffei, S. L. Cohen, B. T. Chait, D. Rabinowitz, R. L. Lallone, S. K. Burley and J. M. Friedman. 1995. Weight-reducing effects of the plasma protein encoded by the obese gene. Science 269:543-546.
- Kim, H. H., M. B. Seol, D. H. Jeon, S. S. Sun, K. H. Kim, Y. J. Choi and M. G. Baik. 2001b. Cloning and expression of lactates dehydrogenase H chain gene in adipose tissues of Korean Cattle. Asian-Aust. J. Anim. Sci. 14:1670-1674.
- Kim, K. H., K. Lee, Y. S. Moon and H. S. Sul. 2001. A cysteinerich adipose tissue-specific secretory factor inhibits adipocyte differentiation. J. Biol. Chem. 276:11252-11256.
- Schafer, R., J. Iyer, E. Iten, A. C. Nirkko, J. Balmer and R. Klemenz. 1991. Molecular and functional analysis of tumorsuppressor genes by transfection. Environ. Health Perspect 93:79-82.
- Smas, C. M., L. Chen and H. S. Sul. 1997. Cleavage of membrane-associated pref-1 generates a soluble inhibitor of adipocyte differentiation. Mol. Cell Biol. 17:977-988.
- Smas, C. M., D. Kachinskas, C. M. Liu, X. Xie, L. K. Dircks and H. S. Sul. 1998. Transcriptional control of the pref-1 gene in 3T3-L1 adipocyte differentiation. Sequence requirement for differentiation-dependent suppression. J. Biol. Chem. 273: 31751-31758.
- Schena, M., D. Shalon, R. W. Davis and P. O. Brown. 1995. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 270:467-470.
- Steppan, C. M., S. T. Bailey, S. Bhat, E. J. Brown, R. R. Banerjee, C. M. Wright, H. R. Patel, R. S. Ahima and M. A. Lazar. 2001. The hormone resistin links obesity to diabetes. Nature 409:307-312.
- Kim, J. H., D. H. Jeon, Y. J. Choi and M. G. Baik. 2001a. Cloning and expression of bovine polyadenylate binding protein 1 cDNA in mammary tissues. Asian-Aust. J. Anim. Sci. 14:771-776.
- Maffei, M., H. Fei, G. H. Lee, C. Dani, P. Leroy, Y. Zhang, R. Proenca, R. Negrel, G. Ailhaud and J. M. Friedman. 1995. Increased expression in adipocytes of ob rna in mice with lesions of the hypothalamus and with mutations at the db locus. Proc. Natl. Acad. Sci. USA 92:6957-6960.
- Hotamisligil, G. S., P. Peraldi, A. Budavari, R. Ellis, M. F. White and B. M. Spiegelman. 1996. Irs-1-mediated inhibition of insulin receptor tyrosine kinase activity in tnf-alpha- and obesity-induced insulin resistance. Science 271:665-668.
- Scherer, P. E., S. Williams, M. Fogliano, G. Baldini and H. F. Lodish. 1995. A novel serum protein similar to c1q, produced exclusively in adipocytes. J. Biol. Chem. 270:26746-26749.
- Zhang, Y., R. Proenca, M. Maffei, M. Barone, L. Leopold and J. M. Friedman. 1994. Positional cloning of the mouse obese gene and its human homologue. Nature 372:425-432.
- Sers, C., U. Emmenegger, K. Husmann, K. Bucher, A. C. Andres and R. Schafer. 1997. Growth-inhibitory activity and downregulation of the class ii tumor-suppressor gene h-rev 107 in tumor cell lines and experimental tumors. J. Cell Biol. 136:935-944.
- DiSepio, D., C. Ghosn, R. L. Eckert, A. Deucher, N. Robinson, M. Duvic, R. A. Chandraratna and S. Nagpal. 1998. Identification and characterization of a retinoid-induced class ii tumor suppressor/growth regulatory gene. Proc. Natl. Acad. Sci. USA 95:14811-14815.
- Holcomb, I. N., R. C. Kabakoff, B. Chan, T. W. Baker, A. Gurney, W. Henzel, C. Nelson, H. B. Lowman, B. D. Wright, N. J. Skelton, G. D. Frantz, D. B. Tumas, F. V. Peale, Jr., D. L. Shelton and C. C. Hebert. 2000. Fizz1, a novel cysteine-rich secreted protein associated with pulmonary inflammation, defines a new gene family. Embo J. 19:4046-4055.
- Hu, E., P. Liang and B. M. Spiegelman. 1996. Adipoq is a novel adipose-specific gene dysregulated in obesity. J Biol Chem 271: 10697-10703.
- Husmann, K., C. Sers, E. Fietze, A. Mincheva, P. Lichter and R. Schafer. 1998. Transcriptional and translational downregulation of h-rev 107, a class ii tumour suppressor gene located on human chromosome 11q11-12. Oncogene, 17:305-1312.
- Lennon, G. G. 2000. High-throughput gene expression analysis for drug discovery. Drug Discov. Today 5:59-66. https://doi.org/10.1016/S1359-6446(99)01448-8
- Shimomura, I., T. Funahashi, M. Takahashi, K. Maeda, K. Kotani, T. Nakamura, S. Yamashita, M. Miura, Y. Fukuda, K. Takemura, K. Tokunaga and Y. Matsuzawa. 1996. Enhanced expression of pai-1 in visceral fat: Possible contributor to vascular disease in obesity. Nat. Med. 2:800-803.
- Huang, S. L., R. Y. Shyu, M. Y. Yeh and S. Y. Jiang. 2000. Cloning and characterization of a novel retinoid-inducible gene 1 (rig1) deriving from human gastric cancer cells. Mol Cell Endocrinol 159:15-24.
- Smas, C. M. and H. S. Sul. 1993. Pref-1, a protein containing egflike repeats, inhibits adipocyte differentiation. Cell 73:725-734.
- Ha, S. H., M. I. Chung, M. G. Baik and Y. J. Choi. 2001. Cloning and expression of b 1-adrenergic receptor genes in adipose tissues from Korean Native Cattle (Hanwoo). Asian-Aust. J. Anim. Sci. 14:13-17.