First Characterization of Sphingomyeline Phosphodiesterase Expression in the Bumblebee, Bombus lantschouensis

lei Han, Shaoyu He, Jie Dong, Ye Wang, Jiaxing Huang, Jie Wu


  The bumblebee (Bombus lantschouensis Vogt) is an important pollinator of wild plants. Sphingomyelin phosphodiesterase (SMPD) is a hydrolase that plays a major role in sphingolipid metabolism reactions. We report the preparation and characterization of a polyclonal antibody for bumblebee SMPD. We then use the polyclonal antiserum to detect the SMPD protein at different development stages and in different tissues. Our results showed that a 1228bp fragment homologous with the B. terrestris SMPD gene was successfully amplified. The molecular weight of the fusion protein was about 70 kDa by SDS-PAGE. An effective polyclonal antibody against SMPD was also obtained from mice and found to have a higher specificity for bumblebee SMPD. Western blotting detection showed that SMPD was expressed at a high level in queen ovaries, although expression was lower in the midgut and venom gland. SMPD expression decreased from the egg stage until the pdd stage. We interpret our results as showing that the development of an effective polyclonal antiserum for the SMPD protein of a bumblebee, which provides a tool for exploring the function of the SMPD gene. In addition, the work has confirmed that SMPD should be considered as an important enzyme during bumblebee egg and larval stages.


Bombus lantschouensis, SMPD gene, gene clone, prokaryotic expression,

Full Text:



Ago, H., Oda, M., Takahashi, M., Tsuge, H., Ochi, S., Katunuma, N., Miyano, M. & Sakurai, J. (2006). Structural basis of the sphingomyelin phosphodiesterase activity in neutral sphingomyelinase from Bacillus cereus. Journal of Biological Chemistry, 281:16157-16167. doi:10.1074/jbc.M601089200

An, J.D., Huang, J.X., Williams, P.H., Wu, J. & Zhou, B.F. (2010). Species diversity and colony characteristics of bumblebees in the Hebei region of North China. Chinese Journal of Applied Ecology, 21: 1542-1550.

Bartelsen, O., Lansmann, S., Nettersheim, M., Lemm, T., Ferlinz, K. & Sandhoff, K. (1998). Expression of recombinant human acid sphingomyelinase in insect Sf21 cells: purification, processing and enzymatic characterization. Journal of Biotechnology, 63: 29-40. doi:10.1016/S0168-1656(98)00070-4

Benoit, J.B., Attardo, G.M., Michalkova, V., Taká?, P., Bohova, J. & Aksoy, S. (2012). Sphingomyelinase activity in mother's milk is essential for juvenile development: a case from lactating tsetse flies. Biology of Reproduction, 87: 1-10. doi: 10.1095/biolreprod.112.100008

Brady, R.O., Kanfer, J.N., Mock, M.B. & Fredrickson, D.S. (1966). The metabolism of sphingomyelin, II. Evidence of an enzymatic deficiency in Niemann-Pick diseae. Proceedings of the National Academy of Sciences, USA 55: 366-369.

Chu, H.Y. & Englund, J.A. (2013). Respiratory syncytial virus disease: prevention and treatment. In: L.J. Anderson & B.S. Graham (Eds.), Challenges and opportunities for respiratory syncytial virus vaccines (pp. 235-258). Berlin Heidelberg: Springer-Verlag.

Dallacqua, R.P., Sim?es, Z.L.P. & Bitondi, M.M.G. (2007). Vitellogenin gene expression in stingless bee workers differing in egg-laying behavior. Insectes Sociaux, 54: 70-76. doi: 10.1007/s00040-007-0913-1

Ding, S.Q., Wang, J., Wang, S.J., Zhang, Y. & Zhao, W. (2006). Echinococcus granulosus 2 heat shock protein 70 gene: construction of recombinant plasmid, prokaryotic expression, purification and characterization. Chinese Journal of Zoonoses, 22: 764-763.

Doll, V.M., Ehling-Schulz, M. & Vogelmann, R. (2013). Concerted action of sphingomyelinase and non-hemolytic enterotoxin in pathogenic Bacillus cereus. PLoS One, 8: e61404-e61404. doi: 10.1371/journal.pone.0061404

Duchateau, M.J. & Velthuis, H.H.W. (1989). Ovarian development and egg laying in workers of Bombus terrestris. Entomologia Experimentalis et Applicata, 51: 199-213. doi:10.1111/j.1570-7458.1989.tb01231.x

Herterving, E., Nilsson, ?., Nyberg, L. & Duan, R.D. (1997). Alkaline sphingomyelinase activity is decreased in human colorectal carcinoma. Cancer, 79: 448-453. doi: 10.1002/(SICI)1097-0142

Hofmann, K., Tomiuk, S., Wolff, G. & Stoffel, W. (2000). Cloning and characterization of the mammalian brain-specific, Mg2+-dependent neutral sphingomyelinase. Proceedings of the National Academy of Sciences, USA, 97: 5895-5900. doi: 10.1073/pnas.97.11.5895

Horinouchi, K., Erlich, S., Perl, D.P., Ferlinz, K., Bisgaier, C.L., Sandhoff, K., Desnick, R.J., Stewart, C.L. & Schuchman, E.H. (1995). Acid sphingomyelinase deficient mice: a model of types A and B Niemann-Pick disease. Nature Genetics, 10: 288-293. doi:10.1038/ng0795-288

Jenkins, R.W., Canals, D. & Hannun, Y.A. (2009). Roles and regulation of secretory and lysosomal acid sphingomyelinase. Cellular Signalling, 21: 836-846. doi:10.1016/j.cellsig.2009.01.026

Kramer, M., Quickert, S., Sponholz, C., Menzel, U., Huse, K., Platzer, M., Bauer, M. & Claus, R.A. (2015). Alternative splicing of SMPD1 in human sepsis. PLoS One, 10(4): e0124503. doi:10.1371/journal.pone.0124503

Kraut, R. (2011). Roles of sphingolipids in Drosophila, development and disease. Journal of Neurochemistry, 116: 764-778. doi: 10.1111/j.1471-4159.2010.07022.x

Kwon, Y.J., Saeed, S. & Duchateau, M.J. (2003). Stimulation of colony initiation and colony development in Bombus terrestris by adding a male pupa: the influence of age and orientation. Apidologie, 34: 429-437. doi: 10.1051/apido:2003039

Liu, Z.Z., Hao, L., Fan, T., He, H.G., You, H.G., Tang, R.C. & Han, C.H. (2011). Construction and identification of prokaryotic expression vector of human TNF-related apoptosis inducing ligand. Progress in Modern Biomedicine, 27: 59-62.

Lynch, A.P., O'Sulliva, F. & Ahearne, M. (2016). The effect of growth factor supplementation on corneal stromal cell phenotype in vitro using a serum-free media. Experimental Eye Research, 151: 26-37. doi: 10.1016/j.exer.2016.07.015

Park, J.H., Choi, J.W., Ju, E.J., Pae, A.N. & Park, K.D. (2015). Antioxidant and anti-inflammatory activities of a natural compound, Shizukahenriol, through Nrf2 activation. Molecules, 20: 15989-16003. doi:10.3390/molecules200915989

Peng, W.J., Huang, J. X., Wu, J. & An, J.D. (2009). Geographic distribution and bionomics of six bumblebee species in North China. Chinese Bulletin of Entomology, 46: 115-120.

Renault, A.D., Starz-Gaiano, M. & Lehmann, R. (2002). Metabolism of sphingosine 1-phosphate and lysophosphatidic acid: a genome wide analysis of gene expression in Drosophila. Mechanisms of Development, 119: S293-S301. doi:10.1016/S0925-4773(03)00131-X

Sabourdy, F., Kedjouar, B., Sorli, S.C., Colié, S., Milhas, D., Salma, Y. & Levade, T. (2008). Functions of sphingolipid metabolism in mammals-Lessons from genetic defects. Biochimica et Biophysica Acta Molecular and Cell Biology of Lipids, 1781: 145-183. doi:10.1016/j.bbalip.2008.01.004

Sjöqvist, U., Hertervig, E., Nilsson, Å., Duan, R.D., Öst, Å., Tribukait, B. & Löfberg, R. (2002). Chronic colitis is associated with a reduction of mucosal alkaline sphingomyelinase activity. Inflammatory Bowel Diseases, 8: 258-263. doi:10.1097/00054725-200207000-00004

Smith, E.L. & Schuchman, E.H. (2008). The unexpected role of acid sphingomyelinase in cell death and the pathophysiology of common diseases. The Faseb Journal, 22: 3419-3431. doi:10.1096/fj.08-108043

Truman, J.P., Al Gadban, M.M., Smith, K.J. & Hammad, S.M. (2011). Acid sphingomyelinase in macrophage biology. Cellular and Molecular Life Sciences, 68: 3293-3305. doi:10.1007/s00018-011-0686-6

Yamada, A., Tsukagoshi, N., Udaka, S., Sasaki, T., Makino, S., Nakamura, S., Little, C., Tomita, M. & Ikezawa, H. (1988). Nucleotide sequence and expression in Escherichia coli of the gene coding for sphingomyelinase of Bacillus cereus. European Journal of Biochemistry, 175: 213-220. doi:10.1111/j.1432-1033.1988.tb14186.x

Yu, Z.J., Ma, X.N. & Zhou, J.H. (2007). A modified method for purification of inclusion bodies proteins in Gel slices. Biotechnology, 17: 46-48.

Zhang, R.L., Meng, J.X., Wen, A.M., Huang, Y.S., Zhou, C.Q., Wang, K. & Chen, X.G. (2009). siRNAs targetting sphingomyelin phosphodiesterase 1 protect mouse oocytes from apoptosis. Journal of Southern Medical University, 29: 2165-2170.

Zhao, W., Su, C., Wu, H.W., Hu, X.M., Shen, L., Wang, R.Z., Ma, L., Chen, S.Z. & Zhang, Z.S. (2000). Construction of recombinant pGEX-6P-1/Sj-FABPc and expression in E.coli. Journal of Tropical Diseases and Parasitology, 12: 261-264.

Zinke, I., Schütz, C.S., Katzenberger, J.D., Bauer, M. & Pankratz, M.J. (2002). Nutrient control of gene expression in Drosophila : microarray analysis of starvation and sugar-dependent response. The Embo Journal, 21: 6162-6173. doi:10.1093/emboj/cdf600



  • There are currently no refbacks.

JCR Impact Factor 2015: 0.702