Molecular Cloning, Characterization and Expression Analysis of Macrohage-Colony Stimulating Factor 2 Gene from Grass Carp (Ctenopharyngodon idellus)

Linyong Du, Lei Qin, Shangnian Wang, Lu Yang, Kun Yang, Xinyan Wang, Hong Zhou

School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, China.

DOI: 10.4236/eng.2013.510B046                

Abstract

Macrophage colony-stimulating factor (CSF-1/M-CSF) is a key factor for the differentiation, growth and survival of monocytes/macrophages and osteoclasts. The functions of M-CSF have been well characterized in mammals. In this study, we have cloned and sequenced the cDNA of M-CSF2 in grass carp. The grass carp M-CSF2 cDNA was 1487 bp in length, containing an open reading frame of 855 bp that encodes 284 amino acids. The deduced protein of grass carp M-CSF2 possessed same domains similar to its mammalian counterparts. Multiple alignments and phylogenetic tree indicate that the grass carp M-CSF2 exhibits close evolutionary relationship with its counterparts in other teleosts. Lastly, the tissue distribution results also showed that the grass carp M-CSF2 transcript was dominantly expressed in head kidney, kidney and spleen in vivo.

Keywords

Grass Carp; M-CSF2; Macrophages; Cloning; Sequence Analysis; Tissue Distribution

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	P. C. Hanington, et al., “Growth Factors of Lower Vertebrates: Characterization of Goldfish (Carassius auratus L.) Macrophage Colony-Stimulating Factor-1,” The Journal of Biological Chemistry, Vol. 282, 2007, pp. 31865-31872. http://dx.doi.org/10.1074/jbc.M706278200
[2]	T. Wang, et al., “Two Macrophage Colony-Stimulating Factor Genes Exist in Fish That Differ in Gene Organization and Are Differentially Expressed,” The Journal of Immunology, Vol. 181, 2008, pp. 3310-3322.
[3]	C. A. Janeway Jr. and R. Medzhitov, “Innate Immune Recognition,” Annual Review of Immunology, Vol. 20, 2002, pp. 197-216. http://dx.doi.org/10.1146/annurev.immunol.20.083001.084359
[4]	F. O. Martinez, et al., “Alternative Activation of Macrophages: An Immunologic Functional Perspective,” Annual Review of Immunology, Vol. 27, 2009, pp. 451-483. http://dx.doi.org/10.1146/annurev.immunol.021908.132532
[5]	V. Chitu and E. R. Stanley, “Colony-Stimulating Factor-1 in Immunity and Inflammation,” Current Opinion in Immunology, Vol. 18, 2006, pp. 39-48. http://dx.doi.org/10.1016/j.coi.2005.11.006
[6]	P. E. Cohen, et al., “Absence of Colony-Stimulating Factor-1 in Osteopetrotic (Csfmop/Csfmop) Mice Results in Male Fertility Defects,” Biology of Reproduction, Vol. 55, 1996, pp. 310-317. http://dx.doi.org/10.1095/biolreprod55.2.310
[7]	P. E. Cohen, et al., “Absence of Colony Stimulating Factor-1 in Osteopetrotic (Csfmop/Csfmop) Mice Disrupts Estrous Cycles and Ovulation,” Biology of Reproduction, Vol. 56, 1997, pp. 110-118. http://dx.doi.org/10.1095/biolreprod56.1.110
[8]	L. R. Rohrschneider, et al., “Growth and Differentiation Signals Regulated by the M-CSF Receptor,” Molecular Reproduction and Development, Vol. 46, 1997, pp. 96- 103. http://dx.doi.org/10.1002/(SICI)1098-2795(199701)46:1<96::AID-MRD15>3.0.CO;2-1
[9]	X. M. Dai, et al., “Targeted Disruption of the Mouse Colony-Stimulating Factor 1 Receptor Gene Results in Osteopetrosis, Mononuclear Phagocyte Deficiency, Increased Primitive Progenitor Cell Frequencies, and Reproductive Defects,” Blood, Vol. 99, 2002, pp. 111-120. http://dx.doi.org/10.1182/blood.V99.1.111
[10]	P. Fixe and V. Praloran, “M-CSF: Haematopoietic Growth Factor or Inflammatory Cytokine?” Cytokine, Vol. 10, 1998, pp. 32-37. http://dx.doi.org/10.1006/cyto.1997.0249
[11]	M. J. Sweet and D. A. Hume, “CSF-1 as a Regulator of Macrophage Activation and Immune Responses,” Archivum Immunologiae et Therapia Experimentalis (Warsz), Vol. 51, 2003, pp. 169-177.
[12]	L. Grayfer, et al., “Macrophage Colony-Stimulating Factor (CSF-1) Induces Pro-Inflammatory Gene Expression and Enhances Antimicrobial Responses of Goldfish (Carassius auratus L.) Macrophages,” Fish and Shellfish Immunology, Vol. 26, 2009, pp. 406-413. http://dx.doi.org/10.1016/j.fsi.2008.12.001
[13]	I. T. Rombel, et al., “ORF-FINDER: A Vector for High-Throughput Gene Identification,” Gene, Vol. 282, 2002, pp. 33-41. http://dx.doi.org/10.1016/S0378-1119(01)00819-8
[14]	J. D. Bendtsen, et al., “Improved Prediction of Signal Peptides: SignalP 3.0,” Journal of Molecular Biology, Vol. 340, 2004, pp. 783-795. http://dx.doi.org/10.1016/S0378-1119(01)00819-8
[15]	M. Ikeda, et al., “Transmembrane Topology Prediction Methods: A Re-Assessment and Improvement by a Consensus Method Using a Dataset of Experimentally-Characterized Transmembrane Topologies,” In Silico Biology, Vol. 2, 2002, pp. 19-33.
[16]	J. Schultz, et al., “SMART, a Simple Modular Architecture Research Tool: Identification of Signaling Domains,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, 1997, pp. 5857-5864. http://dx.doi.org/10.1073/pnas.95.11.5857
[17]	K. Tamura, et al., “MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0,” Molecular Biology and Evolution, Vol. 24, 2007, pp. 1596- 1599. http://dx.doi.org/10.1093/molbev/msm092
[18]	D. A. Hume, et al., “The Mononuclear Phagocyte System Revisited,” Journal of Leukocyte Biology, Vol. 72, pp. 621-627.
[19]	A. Bonnieu, et al., “AUUUA Motifs Are Dispensable for Rapid Degradation of the Mouse c-myc RNA,” Oncogene, Vol. 5, 1990, pp. 1585-1588.
[20]	S. Savant-Bhonsale and D. W. Cleveland, “Evidence for Instability of mRNAs Containing AUUUA Motifs Mediated through Translation-Dependent Assembly of a > 20S Degradation Complex,” Genes & Development, Vol. 6, 1992, pp. 1927-1939. http://dx.doi.org/10.1101/gad.6.10.1927