Direct localisation of molecules in tissue sections of growing antler tips using MALDI imaging
- 298 Downloads
The astonishing growth rate of deer antlers offers a valuable model for the discovery of novel factors and regulatory systems controlling rapid tissue growth. Numerous molecules have been identified in growing antlers using a variety of techniques. However, little is known about the spatial distribution of these molecules in situ. A technique that has the potential to help in this regard is direct proteomic analysis of tissue sections by matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS). The present study applied this technique to spatially map molecules in antler tissue sections. Two protonated molecular ions were selected: m/z 6679 and m/z 6200 corresponding to VEGF and thymosin beta-10, respectively. Superimposition of the respective ion images on to histologically stained samples showed distinct spatial distribution across the antler tissue sections which were consistent with the previous reports using in situ hybridization. Two other molecular ions specifically m/z 8100 and m/z 11,800 were also selected, corresponding to reported masses of urocortin precursor and thioredoxin, respectively. As the spatial distribution of these proteins is not specifically known, MALDI-IMS was used as a potential technique to obtain information on their distribution on antler tips. The presence of all these molecules in deer antlers were further confirmed using LC–MS/MS data. The present study also demonstrated that MALDI-IMS could be further used to image antler sections with an extended ion mass range of up to m/z 45,000, thus potentially increasing the ability to discover the distribution of a larger set of molecules that may play an important role in antler growth. We have thus demonstrated that MALDI-IMS is a promising technique for generating molecular maps with high spatial resolution which can aid in evaluating the function of novel molecules during antler growth.
KeywordsDeer Antler MALDI-IMS Growth centre Proliferation
The authors would like to thank Invermay deer farm crew and Otago Venison Ltd for helping collect antler tissue samples.
C.L., S.DC., S.C., C.M., and J.D conceived and designed the experiments. S.DC., W.W., and C.L performed the experiments. S.DC., W.W., C.L., S.C, and C.M analysed the data. C.L., S.DC., S.C., C.M., W.W., and J.D wrote this paper.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 2.Goss RJ (1983) Deer antlers. Regeneration, function and evolution. Academic Press, New YorkGoogle Scholar
- 4.Li C, Zhao S, Wang W (1988) Biology of deer antler. Chinese Press of Agricultural Sciences, BeijingGoogle Scholar
- 6.Li C (2003) Development of deer antler model for biomedical research. Adv Res Updat 4(2):256–274Google Scholar
- 9.Banks WJ, Newbrey JW (1982) Light microscopic studies of the ossification process in developing antlers. In: Brown RD (ed) Antler Development in Cervidae. Caesar Kleberg Wildl. Res. Inst., KingsvilleGoogle Scholar
- 17.Gyurjan I Jr, Molnar A, Borsy A, Steger V, Hackler L Jr, Zomborszky Z, Papp P, Duda E, Deak F, Lakatos P, Puskas LG, Orosz L (2007) Gene expression dynamics in deer antler: mesenchymal differentiation toward chondrogenesis. Mol Genet Genomics 277(3):221–235. doi: 10.1007/s00438-006-0190-0 CrossRefPubMedGoogle Scholar
- 18.Molnar A, Gyurjan I, Korpos E, Borsy A, Steger V, Buzas Z, Kiss I, Zomborszky Z, Papp P, Deak F, Orosz L (2007) Identification of differentially expressed genes in the developing antler of red deer Cervus elaphus. Mol Genet Genomics 277(3):237–248. doi: 10.1007/s00438-006-0193-x CrossRefPubMedGoogle Scholar
- 21.Borsy A, Podani J, Steger V, Balla B, Horvath A, Kosa JP, Gyurjan I Jr, Molnar A, Szabolcsi Z, Szabo L, Jako E, Zomborszky Z, Nagy J, Semsey S, Vellai T, Lakatos P, Orosz L (2009) Identifying novel genes involved in both deer physiological and human pathological osteoporosis. Mol Genet Genomics 281(3):301–313. doi: 10.1007/s00438-008-0413-7 CrossRefPubMedGoogle Scholar
- 28.Dani FR, Francese S, Mastrobuoni G, Felicioli A, Caputo B, Simard F, Pieraccini G, Moneti G, Coluzzi M, Della TA, Turillazzi S (2008) Exploring proteins in Anopheles gambiae male and female antennae through MALDI mass spectrometry profiling. PLoS One. doi: 10.1371/journal.pone.0002822 PubMedCentralPubMedGoogle Scholar
- 42.Lord E, Clark D, Martin S, Pedersen G, Gray J, Li C (2005) Suttie J Profiling genes expressed in the regenerating tip of red deer (Cervus elaphus) antler. In: Suttie J, Haines S, Li C (eds) Proceedings of the 2nd International Symposium on Antler Science and Product Technology. Queenstown, New Zealand, pp 129–134Google Scholar
- 44.Price JS, Oyajobi BO, Nalin AM, Frazer A, Russell RG, Sandell LJ (1996) Chondrogenesis in the regenerating antler tip in red deer: expression of collagen types I, IIA, IIB, and X demonstrated by in situ nucleic acid hybridization and immunocytochemistry. Dev Dyn 205:332–347CrossRefPubMedGoogle Scholar