Journal of Ocean University of China

, Volume 18, Issue 1, pp 210–218 | Cite as

Ion Transport Signal Pathways Mediated by Neurotransmitter (Biogenic Amines) of Litopenaeus vannamei Under Low Salinity Challenge

  • Luqing PanEmail author
  • Lingjun Si
  • Dongxu Hu


The Pacific white shrimp, Litopenaeus vannamei, is widely farmed in China. Salinity is a major environmental factor that affects its growth and distribution. Crustacean hyperglycemic hormone is verified to participate in ion transport in response to the salinity challenge mediated by endocrine neurotransmitters (biogenic amines, BAs). In the present study, the contents of BAs and expressions of their receptors were detected in gills of Litopenaeus vannamei exposed to low salinity. The intracellular signaling molecules such as cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), 14-3-3 protein, FXYD2 protein and cAMP response element-binding protein (CREB) were detected. The effects of low salinity on the expressions of Na+/K+-ATPase, Na+/K+/2Clco-transporter and Cltransporter and activity of Na+/K+-ATPase were also analyzed. The results showed that dopamine and epinephrine concentrations and their receptor expressions were significantly affected by low salinity. The changes of cAMP and PKA were obvious and the expressions of 14-3-3 and FXYD2 peaked at early stages. However, the expression of CREB was only significantly up-regulated on day 9. The activity and expression of Na+-K+-ATPase (α subunit) reached a peak on day 1. The expressions of Na+/K+/2Clco-transporter and Cltransporter up-regulated obviously. It suggests that BAs can activate the cAMP-PKA pathway, which further acts on the 14-3-3 and FXYD2 proteins, and ultimately improve the activity of Na+/K+-ATPase. Furthermore, after BAs stimulate the cAMP-PKA pathway, PKA phosphorylates the transcription factor CREB and regulates the expressions of ion transport enzymes/ transporters. The results in this study are helpful for understanding the response mechanism of endocrine neurotransmitters on osmoregulation in crustaceans.

Key words

ion regulation Litopenaeus vannamei low salinity neurotransmitter BA receptor 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



This work was supported by State Oceanic Administration Specific Public Project of China (No. 201305005) and the National Natural Science Foundation of China (No. 31072193). We thank all the laboratory members for their technical advice and helpful discussions.


  1. Bradford, M. M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry, 72: 248–254.CrossRefGoogle Scholar
  2. Bray, W. A., Lawrence, A. L., and Leung–Trujillo, J. R., 1994. The effect of salinity on growth and survival of Penaeus vannamei, with observations on the interaction of IHHN virus and salinity. Aquaculture, 122 (2–3): 133–146.CrossRefGoogle Scholar
  3. Buckley, S. J., Fitzgibbon, Q. P., Smith, G. G., and Ventura, T., 2016. In silico prediction of the G–protein coupled receptors expressed during the metamorphic molt of Sagmariasus verreauxi (Crustacea: Decapoda) by mining transcriptomic data: RNA–seq to repertoire. General and Comparative Endocrinology, 228: 111–127.CrossRefGoogle Scholar
  4. Buranajitpirom, D., Asuvapongpatana, S., Weerachatyanukul, W., Wongprasert, K., Namwong, W., Poltana, P., and Withyachumnarnkul, B., 2010. Adaptation of the black tiger shrimp, Penaeus monodon, to different salinities through an excretory function of the antennal gland. Cell and Tissue Research, 340: 481–489.CrossRefGoogle Scholar
  5. Camacho–Jiménez, L., Díaz, F., Muñoz–Márquez, M. E., Farfán, C., Re, A. D., and Ponce–Rivas, E., 2017. Hyperglycemic and osmotic effects of dopamine and recombinant hormone CHHB1 in the Pacific white shrimp Litopenaeus vannamei. Marine and Freshwater Behaviour and Physiology, 50 (1): 67–79.CrossRefGoogle Scholar
  6. Chandrasekar, S., Nich, T., Tripathi, G., Sahu, N. P., Pal, A. K., and Dasgupta, S., 2014. Acclimation of brackish water pearl spot (Etroplus suratensis) to various salinities: Relative changes in abundance of branchial Na+/K+–ATPase and Na+/K+/2Cl–co–transporter in relation to osmoregulatory parameters. Fish Physiology and Biochemistry, 40 (3): 983–996.CrossRefGoogle Scholar
  7. Chang, E. S., and Mykles, D. L., 2011. Regulation of crustacean molting: A review and our perspectives. General and Comparative Endocrinology, 172 (3): 323–330.CrossRefGoogle Scholar
  8. Chen, T., Ren, C., Wang, Y., Gao, Y., Wong, N. K., Zhang, L., and Hu, C., 2016. Crustacean cardioactive peptide (CCAP) of the pacific white shrimp (Litopenaeus vannamei): Molecular characterization and its potential roles in osmoregulation and freshwater tolerance. Aquaculture, 451: 405–412.CrossRefGoogle Scholar
  9. Christensen, J. D., Monaco, M. E., and Lowery, T. A., 1997. An index to assess the sensitivity of Gulf of Mexico species to changes in estuarine salinity regimes. Gulf and Caribbean Research, 9: 219–229.Google Scholar
  10. Chung, J. S., and Webster, S. G., 2006. Binding sites of crustacean hyperglycemic hormone and its second messengers on gills and hindgut of the green shore crab, Carcinus maenas: A possible osmoregulatory role. General and Comparative Endocrinology, 147 (2): 206–213.CrossRefGoogle Scholar
  11. Cortes, V. F., Ribeiro, I. M., Barrabin, H., Alves–Ferreira, M., and Fontes, C. F. L., 2011. Regulatory phosphorylation of FXYD2 by PKC and cross interactions between FXYD2, plasmalemmal Ca–ATPase and Na, K–ATPase. Archives of Biochemistry and Biophysics, 505 (1): 75–82.CrossRefGoogle Scholar
  12. Efendiev, R., Chen, Z., Krmar, R. T., Uhles, S., Katz, A. I., Pedemonte, C. H., and Bertorello, A. M., 2005. The 14–3–3 protein translates the Na+, K+–ATPase a1–subunit phosphorylation signal into binding and activation of phosphoinositide 3–kinase during endocytosis. Journal of Biological Chemistry, 280 (16): 16272–16277.CrossRefGoogle Scholar
  13. Exton, J. H., and Harper, S. C., 1975. Role of cyclic AMP in the actions of catecholamines on hepatic carbohydrate metabolism. Advances in Cyclic Nucleotide Research, 5: 519–532.Google Scholar
  14. Gong, H., Jiang, D. H., Lightner, D. V., Collins, C., and Brock, D., 2004. A dietary modification approach to improve the osmoregulatory capacity of Litopenaeus vannamei cultured in the Arizona desert. Aquaculture Nutrition, 10 (4): 227–236.CrossRefGoogle Scholar
  15. Havird, J. C., Santos, S. R., and Henry, R. P., 2014. Osmoregulation in the Hawaiian anchialine shrimp Halocaridina rubra (Crustacea: Atyidae): Expression of ion transporters, mitochondria–rich cell proliferation and hemolymph osmolality during salinity transfers. Journal of Experimental Biology, 217 (13): 2309–2320.CrossRefGoogle Scholar
  16. Hirose, S., Kaneko, T., Naito, N., and Takei, Y., 2003. Molecular biology of major components of chloride cells. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 136 (4): 593–620.CrossRefGoogle Scholar
  17. Jayasundara, N., Towle, D. W., Weihrauch, D., and Spanings–Pierrot, C., 2007. Gill–specific transcriptional regulation of Na+/K+–ATPase alpha–subunit in the euryhaline shore crab Pachygrapsus marmoratus: Sequence variants and promoter structure. Journal of Experimental Biology, 210: 2070–2081.CrossRefGoogle Scholar
  18. Kaeodee, M., Pongsomboon, S., and Tassanakajon, A., 2011. Expression analysis and response of Penaeus monodon 14–3–3 genes to salinity stress. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 159 (4): 244–251.CrossRefGoogle Scholar
  19. Leone, F. A., Garçon, D. P., Lucena, M. N., Faleiros, R. O., Azevedo, S. V., Pinto, M. R., and McNamara, J. C., 2015. Gill–specific (Na+, K+)–ATPase activity and a–subunit mRNA expression during low–salinity acclimation of the ornate blue crab Callinectes ornatus (Decapoda, Brachyura). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 186: 59–67.CrossRefGoogle Scholar
  20. Li, E., Wang, S., Li, C., Wang, X., Chen, K., and Chen, L., 2014. Transcriptome sequencing revealed the genes and pathways involved in salinity stress of Chinese mitten crab, Eriocheir sinensis. Physiological Genomics, 46: 177–190.CrossRefGoogle Scholar
  21. Li, E., Wang, X., Chen, K., Xu, C., Qin, J. G., and Chen, L., 2017. Physiological change and nutritional requirement of Pacific white shrimp Litopenaeus vannamei at low salinity. Reviews in Aquaculture, 9 (1): 57–75.CrossRefGoogle Scholar
  22. Liu, H. Y., Pan, L. Q., and Zheng, D. B., 2008. Injection of biogenic amines modulates osmoregulation of Litopenaeus vannamei: Response of hemolymph osmotic pressure, ion concentration and osmolality effectors. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 151 (2): 191–197.CrossRefGoogle Scholar
  23. Liu, H. Y., Pan, L. Q., and Zheng, D. B., 2009. Effects of injection of biogenic amines on expression of gill related ion transporter mRNA and a–subunit protein in Litopenaeus vannamei. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 154 (1): 29–36.CrossRefGoogle Scholar
  24. Lohrmann, D. M., and Kamemoto, F. I., 1987. The effect of dibutyryl cAMP on sodium uptake by isolated perfused gills of callinectes sapidus. General and Comparative Endocrinology, 65: 300–305.CrossRefGoogle Scholar
  25. Lonze, B. E., and Ginty, D. D., 2002. Function and regulation of CREB family transcription factors in the nervous system. Neuron, 35 (4): 605–623.CrossRefGoogle Scholar
  26. McNamara, J. C., and Faria, S. C., 2012. Evolution of osmoregulatory patterns and gill ion transport mechanisms in the decapod Crustacea: A review. Journal of Comparative Physiology B, 182 (8): 997–1014.CrossRefGoogle Scholar
  27. Moorman, B. P., Inokuchi, M., Yamaguchi, Y., Lerner, D. T., Grau, E. G., and Seale, A. P., 2014. The osmoregulatory effects of rearing Mozambique tilapia in a tidally changing salinity. General and Comparative Endocrinology, 207: 94–102.CrossRefGoogle Scholar
  28. Morris, S., 2001. Neuroendocrine regulation of osmoregulation and the evolution of air–breathing in decapod crustaceans. Journal of Experimental Biology, 204: 979–989.Google Scholar
  29. Neve, K. A., Seamans, J. K., and Trantham–Davidson, H., 2004. Dopamine receptor signaling. Journal of Receptors and Signal Transduction, 24 (3): 165–205.CrossRefGoogle Scholar
  30. Pan, L., Liu, H., and Zhao, Q., 2014. Effect of salinity on the biosynthesis of amines in Litopenaeus vannamei and the expression of gill related ion transporter genes. Journal of Ocean University of China, 13 (3): 453–459.CrossRefGoogle Scholar
  31. Pan, L. Q., Luan, Z. H., and Jin, C. X., 2006. Effects of Na+/K+ and Mg2+/Ca2+ ratios in saline groundwaters on Na+–K+–ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae. Aquaculture, 261: 1396–1402.CrossRefGoogle Scholar
  32. Pfaffl, M. W., 2001. A new mathematical method for relative quantification in real–time RT–PCR. Nucleic Acids Research, 29: 2002–2007.CrossRefGoogle Scholar
  33. Post, R. L., and Jolly, P. C., 1957. The linkage of sodium, potassium, and ammonium active transport across the human erythrocyte membrane. Biochimica et Biophysica Acta, 25: 118–128.CrossRefGoogle Scholar
  34. Qin, K., Sethi, P. R., and Lambert, N. A., 2008. Abundance and stability of complexes containing inactive G protein–coupled receptors and G proteins. The FASEB Journal, 22 (8): 2920–2927.CrossRefGoogle Scholar
  35. Rasmussen, R., 2001. Quantification on the LightCycler. In: Rapid Cycle Real–Time PCR Methods and Applications. Meuer, M. S., ed., Springer, Heidelberg, 21–34.CrossRefGoogle Scholar
  36. Robles, J., Charmantier, G., Boulo, V., Lizarraga–Valdez, J., Enriquez–Paredes, L. M., and Giffard–Mena, I., 2014. Osmoregulation pattern and salinity tolerance of the white shrimp Litopenaeus vannamei (Boone, 1931) during post–embryonic development. Aquaculture, 422: 261–267.CrossRefGoogle Scholar
  37. Ruat, M., Traiffort, E., Leurs, R., Tardivel–Lacombe, J., Diaz, J., Arrang, J. M., and Schwartz, J. C., 1993. Molecular cloning, characterization, and localization of a high–affinity serotonin receptor (5–HT7) activating cAMP formation. Proceedings of the National Academy of Sciences, 90 (18): 8547–8551.CrossRefGoogle Scholar
  38. Saoud, I. P., Davis, D. A., and Rouse, D. B., 2003. Suitability studies of inland well waters for Litopenaeus vannamei culture. Aquaculture, 217 (1–4): 373–383.CrossRefGoogle Scholar
  39. Shu, M. A., Long, C., Dong, W. R., Zhang, P., Xu, B. P., and Guo, X. L., 2015. The full–length cDNA cloning and expression profiles of 14–3–3 genes from the mud crab Scylla paramamosain Estampador, 1949. Crustaceana, 88 (10–11): 1065–1078.CrossRefGoogle Scholar
  40. Silva, E. C., Masui, D. C., Furriel, R. P., McNamara, J. C., Barrabin, H., Scofano, H. M., Perales, J., Teixeira–Ferreira, A., Leone, F. A., and Fontes, C. F. L., 2012. Identification of a crab gill FXYD2 protein and regulation of crab microsomal Na,KATPase activity by mammalian FXYD2 peptide. Biochimica et Biophysica Acta (BBA)–Biomembranes, 1818: 2588–2597.CrossRefGoogle Scholar
  41. Sommer, M. J., and Mantel, H. L., 1988. Effect of dopamine, cyclic AMP, and pericardial organs on sodium uptake and Na+/K+–ATPase activity in gills of the green crab Carcinus maenas (L). Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 248: 272–277.CrossRefGoogle Scholar
  42. Sommer, M. J., and Mantel, L. H., 1991. Effects of dopamine and acclimation to reduced salinity on the concentration of cyclic AMP in the gills of the green crab, Carcinus maenas (L). General and Comparative Endocrinology, 82 (3): 364–368.CrossRefGoogle Scholar
  43. Spanings–Pierrot, C., Soyez, D., Van Herp, F., Gompel, M., Skaret, G., Grousset, E., and Charmantier, G., 2000. Involvement of crustacean hyperglycemic hormone in the control of gill ion transport in the crab Pachygrapsus marmoratus. General and Comparative Endocrinology, 119 (3): 340–350.CrossRefGoogle Scholar
  44. Tierney, E. P., Tulac, S., Huang, J., and Giudice, L., 2003. Activation of the protein kinase a pathway during decidualization of human endometrial stromal cells reveals sequential categorical gene regulation. Fertility and Sterility, 80: 285.CrossRefGoogle Scholar
  45. Trausch, G., Forget, M. C., and Devos, P., 1989. Biominesstimulated phosphorylation and Na+/K+–ATPase in the gills of the Chinese crab Eriocheir sinensis. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 94: 487–492.CrossRefGoogle Scholar
  46. Tsai, J. R., and Lin, H. C., 2007. V–type H+–ATPase and Na+, K+–ATPase in the gills of 13 euryhaline crabs during salinity acclimation. Journal of Experimental Biology, 210 (4): 620–627.CrossRefGoogle Scholar
  47. Vanhoenacker, P., Haegeman, G., and Leysen, J. E., 2000. 5–HT7 receptors: Current knowledge and future prospects. Trends in Pharmacological Sciences, 21 (2): 70–77.CrossRefGoogle Scholar
  48. Wanna, W., Thipwong, J., Mahakaew, W., and Phongdara, A., 2012. Identification and expression analysis of two splice variants of the 14–3–3 epsilon from Litopenaeus vannamei during WSSV infections. Molecular Biology Reports, 39 (5): 5487–5493.CrossRefGoogle Scholar
  49. Wang, L. U., and Chen, J. C., 2005. The immune response of white shrimp Litopenaeus vannamei and its susceptibility to Vibrio alginolyticus at different salinity levels. Fish & Shellfish Immunology, 18 (4): 269–278.CrossRefGoogle Scholar
  50. Wanga, J. M., Zhangb, Y. M., and Wanga, D. H., 2008. Integrative physiology of plateau pikas and root voles on the Qinghai–Tibetan plateau. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 148 (4): 465.Google Scholar
  51. Watts, S. A., Yeh, E. W., and Henry, R. P., 1996. Hypoosmotic stimulation of ornithine decarboxylase activity in the brine shrimp Artemia franciscana. Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 274 (1): 15–22.CrossRefGoogle Scholar
  52. Webster, S. G., Keller, R., and Dircksen, H., 2012. The CHHsuperfamily of multifunctional peptide hormones controlling crustacean metabolism, osmoregulation, moulting, and reproduction. General and Comparative Endocrinology, 175 (2): 217–233.CrossRefGoogle Scholar
  53. Zhao, Q., Pan, L., Ren, Q., Wang, L., and Miao, J., 2016. Effect of salinity on regulation mechanism of neuroendocrine–immunoregulatory network in Litopenaeus vannamei. Fish & Shellfish Immunology, 49: 396–406.CrossRefGoogle Scholar
  54. Zhen, J. L., Chang, Y. N., Qu, Z. Z., Fu, T., Liu, J. Q., and Wang, W. P., 2016. Luteolin rescues pentylenetetrazole–induced cognitive impairment in epileptic rats by reducing oxidative stress and activating PKA/CREB/BDNF signaling. Epilepsy & Behavior, 57: 177–184.CrossRefGoogle Scholar
  55. Zhu, B., Wang, D., Peng, T., Wang, L., Wei, G., and Liu, C., 2014. Characterization and function of a gene Pc 14–3–3 isoform from red crayfish, Procambarus clarkii. Pakistan Journal of Zoology, 46 (1): 107–113.Google Scholar
  56. Xiao, Y. C., Chen, J., Xie, C. Y., Peng, T., Liu, Y., and Wang, W. N., 2017. A diet of fructose–enriched Artemia improves the response of juvenile Litopenaeus vannamei shrimp to acute low–salinity challenge. Aquaculture Research, 48 (7): 3935–3949.CrossRefGoogle Scholar
  57. Xu, C., Li, E., Liu, Y., Wang, X., Qin, J. G., and Chen, L., 2017. Comparative proteome analysis of the hepatopancreas from the Pacific white shrimp Litopenaeus vannamei under longterm low salinity stress. Journal of Proteomics, 162: 1–10.CrossRefGoogle Scholar

Copyright information

© Science Press, Ocean University of China and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The Key Laboratory of Mariculture of Ministry of EducationOcean University of ChinaQingdaoChina

Personalised recommendations