Molecular characterization of the <em>netrin-1 UNC-5</em> receptor in <em>Lucilia sericata</em> larvae

Tahereh Karamzadeh; Hamzeh Alipour; Marziae Shahriari-Namadi; Abbasali Raz; Kourosh Azizi; Masoumeh Bagheri; Mohammad D. Moemenbellah-Fard; Tahereh Karamzadeh; Hamzeh Alipour; Marziae Shahriari-Namadi; Abbasali Raz; Kourosh Azizi; Masoumeh Bagheri; Mohammad D. Moemenbellah-Fard

doi:10.3934/genet.2019.3.46

AIMS Genetics

2019, Volume 6, Issue 3: 46-54. doi: 10.3934/genet.2019.3.46

Previous Article Next Article

Research article

Molecular characterization of the netrin-1 UNC-5 receptor in Lucilia sericata larvae

1.
Research Center for Health Sciences, Institute of Health, Shiraz University of Medical Sciences, Shiraz, Iran
2.
Department of Medical Entomology, School of Health, Shiraz University of Medical Sciences, Shiraz, Iran
3.
Malaria and Vector Research Group, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

Received: 19 June 2019 Accepted: 02 August 2019 Published: 08 August 2019

Larval therapy with Lucilia sericata is a promising strategy in wound healing. Axon guidance molecules play vital roles during the development of the nervous system and also regulate the capacity of neuronal restoration in wound healing. Netrin-1, one of the proteins that larvae secrete, plays a useful role in cell migration and nerve tissue regeneration. The UNC-5 receptor combines with a netrin-1 signal and transmits the signal from one side of the membrane to the other side, initiating a change in cell activity. In the current study, we identified the full length of the UNC-5 receptor mRNA in L. sericata using different sets of primers, including exon junction and specific region primers. The coding sequence (CDS) of the UNC-5 receptor was sequenced and identified to include 633 base-pair nucleic acids, and BLAST analysis on its nucleotide sequence revealed 96% identity with the Lucilia cuprina netrin-1 UNC-5 receptor. The protein residue included 210 amino acids (aa) and coded for a protein with 24 kD weight. This gene lacked the signal peptide. Furthermore, the UPA domain is conserved in UNC-5. It lied at the interval of 26–131 aa. We identified the CDS of netrin-1 UNC-5 receptor in L. sericata. It could be applied to research activities implementing a new essential component design in wound healing.
- molecular profile,
- Netrin-1 UNC-5 receptor,
- Lucilia sericata,
- blowfly larvae
Citation: Tahereh Karamzadeh, Hamzeh Alipour, Marziae Shahriari-Namadi, Abbasali Raz, Kourosh Azizi, Masoumeh Bagheri, Mohammad D. Moemenbellah-Fard. Molecular characterization of the netrin-1 UNC-5 receptor in Lucilia sericata larvae[J]. AIMS Genetics, 2019, 6(3): 46-54. doi: 10.3934/genet.2019.3.46

Related Papers:

Abstract

Larval therapy with Lucilia sericata is a promising strategy in wound healing. Axon guidance molecules play vital roles during the development of the nervous system and also regulate the capacity of neuronal restoration in wound healing. Netrin-1, one of the proteins that larvae secrete, plays a useful role in cell migration and nerve tissue regeneration. The UNC-5 receptor combines with a netrin-1 signal and transmits the signal from one side of the membrane to the other side, initiating a change in cell activity. In the current study, we identified the full length of the UNC-5 receptor mRNA in L. sericata using different sets of primers, including exon junction and specific region primers. The coding sequence (CDS) of the UNC-5 receptor was sequenced and identified to include 633 base-pair nucleic acids, and BLAST analysis on its nucleotide sequence revealed 96% identity with the Lucilia cuprina netrin-1 UNC-5 receptor. The protein residue included 210 amino acids (aa) and coded for a protein with 24 kD weight. This gene lacked the signal peptide. Furthermore, the UPA domain is conserved in UNC-5. It lied at the interval of 26–131 aa. We identified the CDS of netrin-1 UNC-5 receptor in L. sericata. It could be applied to research activities implementing a new essential component design in wound healing.

Acknowledgments

We appreciate colleagues in the Department of Medical Entomology, School of Health, for their collaboration in this research activity. This project was funded by the Shiraz University of Medical Sciences (SUMS), Shiraz, Iran [Grant #: 1396-01-04-15875] allocated to H.A. It was part of an MSc thesis in Medical Entomology and Vector Control solicited to the student, Ms. T. Karamzadeh.

References

[1]	Malekian A, Esmaeeli Djavid Gh, Akbarzadeh K, et al. (2019) Efficacy of maggot therapy on staphylococcus aureus and pseudomonas aeruginosa in diabetic foot ulcers: A randomized controlled trial. J Wound Ostomy Continence Nurs 46: 25–29. doi: 10.1097/WON.0000000000000496
[2]	Iorio V, Troughton LD, Hamill KJ (2015) Laminins: Roles and utility in wound repair. Adv Wound Care 4: 250–263. doi: 10.1089/wound.2014.0533
[3]	Mitchell KJ, Doyle JL, Serafini T, et al. (1996) Genetic analysis of Netrin genes in Drosophila: Netrins guide CNS commissural axons and peripheral motor axons. Neuron17: 203–215.
[4]	Rajasekharan S, Kennedy TE (2009) The netrin protein family. Genome Biol 10: 239. doi: 10.1186/gb-2009-10-9-239
[5]	Poon VY, Klassen MP, Shen K (2008) UNC-6/netrin and its receptor UNC-5 locally exclude presynaptic components from dendrites. Nature 455: 669. doi: 10.1038/nature07291
[6]	Stein E, Zou Y, Poo M-m, et al. (2001) Binding of DCC by netrin-1 to mediate axon guidance independent of adenosine A2B receptor activation. Science 291: 1976–1982. doi: 10.1126/science.1059391
[7]	Bashaw GJ, Goodman CS (1999) Chimeric axon guidance receptors: The cytoplasmic domains of slit and netrin receptors specify attraction versus repulsion. Cell 97: 917–926. doi: 10.1016/S0092-8674(00)80803-X
[8]	Liu Y, Bhowmick T, Liu Y, et al. (2018) Structural basis for Draxin-Modulated Axon guidance and fasciculation by Netrin-1 through DCC. Neuron 97: 1261–1267. doi: 10.1016/j.neuron.2018.02.010
[9]	Xing Y, Lai J, Liu X, et al. (2017) Netrin-1 restores cell injury and impaired-angiogenesis in vascular endothelial cells. J Mol Endocrinol: JME-16-0239.
[10]	Howard LJ, Brown HE, Wadsworth BC, et al. (2017) Midline axon guidance in the Drosophila embryonic central nervous system, In: Seminars in Cell & Developmental Biology, Elsevier.
[11]	Hand RA, Kolodkin AL (2017) Netrin-mediated axon guidance to the CNS midline revisited. Neuron 94: 691–693. doi: 10.1016/j.neuron.2017.05.012
[12]	Llambi F, Causeret F, Bloch‐Gallego E, et al. (2001) Netrin‐1 acts as a survival factor via its receptors UNC5H and DCC. EMBO J 20: 2715–2722.
[13]	Mehlen P, Delloye-Bourgeois C, Chédotal A (2011) Novel roles for Slits and netrins: Axon guidance cues as anticancer targets? Nature Rev Cancer 11: 188. doi: 10.1038/nrc3005
[14]	Wang W, Reeves WB, Ramesh G (2009) Netrin-1 increases proliferation and migration of renal proximal tubular epithelial cells via the UNC5B receptor. Am J Physiol Renal Physiol 296: F723–F9. doi: 10.1152/ajprenal.90686.2008
[15]	Ylivinkka I, Keski-Oja J, Hyytiäinen M (2016) Netrin-1: A regulator of cancer cell motility? Eur J Cell Biol 95: 513–520. doi: 10.1016/j.ejcb.2016.10.002
[16]	Ke X, Li Q, Xu L, et al. (2015) Netrin-1 overexpression in bone marrow mesenchymal stem cells promotes functional recovery in a rat model of peripheral nerve injury. J Biomed Res 29: 380.
[17]	Eming SA, Brachvogel B, Odorisio T, et al. (2007) Regulation of angiogenesis: Wound healing as a model. Progress Histochem Cytochem 42: 115–170. doi: 10.1016/j.proghi.2007.06.001
[18]	Park KW, Crouse D, Lee M, et al. (2004) The axonal attractant Netrin-1 is an angiogenic factor. Proc Nat Acad Sc i101: 16210–16215.
[19]	Sherman RA, Cooper EL (2018) Biotherapy: Medicinal maggots and invertebrate immunology from the Clinician's perspective. Adv Comp Immunol, Springer: 991–995.
[20]	Ramkhelawon B, Hennessy EJ, Ménager M, et al. (2014) Netrin-1 promotes adipose tissue macrophage retention and insulin resistance in obesity. Nature Med 20: 377. doi: 10.1038/nm.3467
[21]	Hadi T, Boytard L, Silvestro M, et al. (2018) Macrophage-derived netrin-1 promotes abdominal aortic aneurysm formation by activating MMP3 in vascular smooth muscle cells. Nature Commun 9: 5022. doi: 10.1038/s41467-018-07495-1
[22]	Fournier AE, Strittmatter SM (2001) Repulsive factors and axon regeneration in the CNS. Curr Opin Neurobiol 11: 89–94. doi: 10.1016/S0959-4388(00)00178-1
[23]	Mumcuoglu KY, Miller J, Mumcuoglu M, et al. (2001) Destruction of bacteria in the digestive tract of the maggot of Lucilia sericata (Diptera: Calliphoridae). J Med Entomol 38: 161–166.
[24]	Alipour H, Raz A, Zakeri S, et al. (2017) Molecular characterization of matrix metalloproteinase-1 (MMP-1) in Lucilia sericata larvae for potential therapeutic applications. Electron J Biotechnol 29: 47–56. doi: 10.1016/j.ejbt.2017.06.007
[25]	Alipour H, Raz A, Zakeri S (2016) Therapeutic applications of collagenase (metalloproteases): A review. Asian Pac J Trop Biomed 6: 975–981. doi: 10.1016/j.apjtb.2016.07.017
[26]	Alipour H, Raz A, Djadid ND, et al. (2017) Lucilia sericata collagenase. Google Pat.
[27]	Akbarzadeh K, Rafinejad J, Alipour H, et al. (2012) Human myiasis in Fars province, Iran. Southeast Asian J Trop Med Public Health 43: 1205.
[28]	Rafinejad J, Akbarzadeh K, Rassi Y, et al. (2014) Traumatic myiasis agents in Iran with introducing of new dominant species, Wohlfahrtia magnifica (Diptera: Sarcophagidae). Asian Pacific J Trop Biomed 4: 451–455. doi: 10.12980/APJTB.4.2014C1029
[29]	Evans TA, Bashaw GJ (2012) Slit/Robo-mediated axon guidance in Tribolium and Drosophila: Divergent genetic programs build insect nervous systems. Dev Biol 1: 266–278
[30]	Čeřovský V, Žďárek J, Fučík V, et al. (2010) Lucifensin, the long-sought antimicrobial factor of medicinal maggots of the blowfly Lucilia sericata. Cell Mol Life Sci 67: 455–466. doi: 10.1007/s00018-009-0194-0
[31]	Telford G, Brown A, Seabra R, et al. (2010) Degradation of eschar from venous leg ulcers using a recombinant chymotrypsin from Lucilia sericata. Br J Dermatol 163: 523–531. doi: 10.1111/j.1365-2133.2010.09854.x
[32]	Feinstein J, Ramkhelawon B (2017) Netrins & Semaphorins: Novel regulators of the immune response. BBA-Mol Basis Dis 1863: 3183–3189. doi: 10.1016/j.bbadis.2017.09.010
[33]	Popel AS, Karagiannis ED (2015) Compositions having antiangiogenic activity and uses thereof. Google Pat .
[34]	Mediero A, Wilder T, Ramkhelawon B, et al. (2016) Netrin-1 and its receptor Unc5b are novel targets for the treatment of inflammatory arthritis. FASEB J 30: 3835–3844. doi: 10.1096/fj.201600615R
[35]	Maruyama K, Takemura N, Martino MM, et al. (2017) Netrins as prophylactic targets in skeletal diseases: A double-edged sword? Pharmacol Res 122: 46–52. doi: 10.1016/j.phrs.2017.05.011
[36]	Huttenlocher A, Poznansky MC (2008) Reverse leukocyte migration can be attractive or repulsive. Trends Cell Biol 18: 298–306.
[37]	Vigl B, Aebischer D, Nitschké M, et al. (2011) Tissue inflammation modulates gene expression of lymphatic endothelial cells and dendritic cell migration in a stimulus-dependent manner. Blood: blood-2010-12-326447.
[38]	Rőszer T (2015) Understanding the mysterious M2 macrophage through activation markers and effector mechanisms. Med Inflammation.

Reader Comments

Your name:*

Email:*
© 2019 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)