Veterinary Research Forum
Vet Res Forum

Conventional histomorphometry and fast free of acrylamide clearing tissue visualization of sciatic nerve in chicken (Gallus domesticus)

Document Type : Original Article

Authors

Abdolrasoul Malekpour 1, 2
Farhad Rahmanifar 3

1 DVM Graduate, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

2 Legal Medicine Research Center, Legal Medicine Organization, Tehran, Iran

3 Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

https://doi.org/10.30466/vrf.2019.98930.2361
Abstract
Histomorphometry and use of the fast free of acrylamide clearing tissue (FACT) protocol were studied on the sciatic nerve in chicken (Gallus domesticus). In the first part of the study, the sciatic nerves of 20 chickens of four age groups (7, 14, 26 and 40 days) were studied (n=5 birds per age class). Their sciatic nerve samples were stained with Hematoxylin and Eosin and Masson's trichrome and were histomorphometrically evaluated. In the second part of the study, FACT protocol was applied on the sciatic nerve of a 26 days old chicken. After clearing of 1.00 mm-thick sciatic nerve sections, they were immunolabelled using Hoechst for nuclei staining and recorded by a Z-stack motorized fluorescent microscope. In the conventional histo-morphometry, the epineurium, perineurium and endoneurium were thicker and the nerve bundle diameter was bigger in the left sciatic nerve of chicken of all age groups compared to the right sciatic nerve. On the contrary, the axon diameter and the myelinated nerve fiber diameter were bigger, the myelin sheath was thicker, the nodes of Ranvier intervals were higher and the density of myelinated nerve fibers was also higher in the right sciatic nerve compared to the left one. In conclusion, histomorphometric parameters in the left and right sciatic nerve during chicken growth were significantly different. Furthermore, the FACT protocol could be used for the 3D imaging of the chicken sciatic nerve and its immunostained evaluation.

Keywords

3D imaging
Chicken
FACT
Histomorphometric study
Sciatic nerve
  1. Dodt H-U, Leischner U, Schierloh A, et al. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat Methods 2007; 4(4):331-336.
  2. Tomer R, Ye L, Hsueh B, et al. Advanced CLARITY for rapid and high-resolution imaging of intact tissues. Nat Protoc 2014; 9(7):1682-1697.
  3. Chung K, Wallace J, Kim SY, et al. Structural and molecular interrogation of intact biological systems. Nature 2013; 497(7449):332-337.
  4. Yang B, Treweek JB, Kulkarni RP, et al. Single-cell phenotyping within transparent intact tissue through whole-body clearing. Cell 2014; 158(4):945-958.
  5. Murray E, Cho JH, Goodwin D, et al. Simple, scalable proteomic imaging for high-dimensional profiling of intact systems. Cell 2015; 163(6):1500-1514.
  6. Liu AKL, Lai HM, Chang RC-C, et al. Free of acrylamide sodium dodecyl sulphate (SDS)‐based tissue clearing (FASTClear): a novel protocol of tissue clearing for three‐dimensional visualization of human brain tissues. Neuropathol Appl Neurobiol 2017;43(4): 346-351.
  7. Xu N, Tamadon A, Liu Y, et al. Fast free-of-acrylamide clearing tissue (FACT)- an optimized new protocol for rapid, high-resolution imaging of three-dimensional brain tissue. Scientific Reports 2017; 7(1):9895. doi:10.1038/s41598-017-10204-5.
  8. Oliveira FS, Nessler RA, Castania JA, et al. Ultra-structural and morphometric alterations in the aortic depressor nerve of rats due to long term experimental diabetes: effects of insulin treatment. Brain Res. 2013; 1491:197-203.
  9. Sanada LS, Tavares MR, Neubern MCM, et al. Can Wistar rats be used as the normotensive controls for nerve morphometry investigations in spontaneously hypertensive rats (SHR)? Acta Cir Bras 2011; 26(6):514-520.
  10. Dolapchieva S. Computer-assisted morphometric study on the postnatal growth of axon size and myelin thickness in the ventral and dorsal roots of the rabbit. Ann Anat 2004; 186(1):61-68.
  11. Ronchi G, Jager SB, Vaegter CB, et al. Discrepancies in quantitative assessment of normal and regenerated peripheral nerve fibers between light and electron microscopy. J Peripher Nerv Syst 2014; 19(3):224-233.
  12. Barreiros VCP, Dias FJ, Iyomasa MM, et al. Morphological and morphometric analyses of crushed sciatic nerves after application of a purified protein from natural latex and hyaluronic acid hydrogel. Growth Factors 2014; 32(5):164-170.
  13. Kemp J, Després O, Pebayle T, et al. Differences in age-related effects on myelinated and unmyelinated peripheral fibres: a sensitivity and evoked potentials study. Eur J Pain 2014; 18(4):482-488.
  14. Ikeda M, Oka Y. The relationship between nerve conduction velocity and fiber morphology during peripheral nerve regeneration. Brain Beha 2012; 2(4):382-390.
  15. Fischer AH, Jacobson KA, Rose J, et al. Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc 2008; 2008:pdb.prot4986. doi: 10.1101/pdb.prot4986.
  16. Foot NC. The Masson trichrome staining methods in routine laboratory use. Stain Technol 1933; 8(3): 101-110.
  17. Mohammad Rezazadeh F, Saedi S, Rahmanifar F, et al. Fast free of acrylamide clearing tissue (FACT) for clearing, immunolabelling and three‐dimensional imaging of partridge tissues. Microsc Res Tech 2018; 81(12):1374-1382.
  18. Sta M, Cappaert NLM, Ramekers D, et al. The functional and morphological characteristics of sciatic nerve degeneration and regeneration after crush injury in rats. J Neurosci Methods 2014; 222:189-198.
  19. Christensen MB, Tresco PA. Differences exist in the left and right sciatic nerves of naïve rats and cats. Anat Rec (Hoboken). 2015; 298(8):1492-1501.
  20. Vujaskovic Z, Gillette SM, Powers BE, et al. Effects of intraoperative hyperthermia on canine sciatic nerve: histopathologic and morphometric studies. Int J Hyperthermia 1994; 10(6):845-855.
  21. Perge JA, Niven JE, Mugnaini E, et al. Why do axons differ in caliber? J Neurosci. 2012; 32(2):626-638.
  22. Muglia U, Vita G, Laura R, et al. Morphometric comparison between contralateral sciatic nerves in the male and female rabbit. Anat Histol Embryol 1997; 26(2):147-150.
  23. Dolapchieva S, Eggers R, Kühnel W. Automatic image analysis of the postnatal growth of axons and myelin sheaths in the tibial and peroneal nerves of the rabbit. Ann Anat 2000; 182(2):133-142.
  24. Hartline DK, Colman DR. Rapid conduction and the evolution of giant axons and myelinated fibers. Curr Biol 2007; 17(1):R29-35.
  25. Pannese E. Morphological changes in nerve cells during normal aging. Brain Struct Funct. 2011; 216(2):85-89.
  26. Susaki EA, Tainaka K, Perrin D, et al. Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis. Cell 2014; 157(3):726-739.
  27. Hama H, Kurokawa H, Kawano H, et al. Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain. Nat Neurosci 2011; 14(11):1481-1488.
  28. Zheng H, Rinaman L. Simplified CLARITY for visualizing immunofluorescence labeling in the developing rat brain. Brain Struct Funct 2016; 221(4): 2375-2383.
  29. Liang H, Schofield E, Paxinos G. Imaging serotonergic fibers in the mouse spinal cord using the CLARITY/CUBIC technique. J Vis Exp 2016(108): 53673. doi: 10.3791/53673
  30. Saboor F, Reckmann AN, Tomczyk CUM, et al. Nestin-expressing vascular wall cells drive development of pulmonary hypertension. Eur Respir J 2016; 47(3): 876-888.
  31. Ding Y, Lee J, Ma J, et al. Light-sheet fluorescence imaging to localize cardiac lineage and protein distribution. Sci Rep 2017; 7:42209. doi: 10.1038/ srep42209.
  32. Font-Burgada J, Shalapour S, Ramaswamy S, et al. Hybrid periportal hepatocytes regenerate the injured liver without giving rise to cancer. Cell 2015; 162(4): 766-779.
  33. Unnersjö-Jess D, Scott L, Blom H, et al. Super-resolution stimulated emission depletion imaging of slit diaphragm proteins in optically cleared kidney tissue. Kidney Int 2016; 89(1):243-247.
  34. Neckel PH, Mattheus U, Hirt B, et al. Large-scale tissue clearing (PACT): Technical evaluation and new perspectives in immunofluorescence, histology, and ultrastructure. Sci Rep 2016; 6:34331. doi: 10.1038/ srep34331.
  35. Muzumdar MD, Dorans KJ, Chung KM, et al. Clonal dynamics following p53 loss of heterozygosity in Kras-driven cancers. Nat Commun 2016; 7:12685. doi: 10.1038/ncomms12685.
  36. Murray K, Godinez DR, Brust-Mascher I, et al. Neuroanatomy of the spleen: Mapping the relationship between sympathetic neurons and lymphocytes. PLoS One 2017; 12(7):e0182416. doi:10.1371/journal. pone.0182416.
  37. Lloyd-Lewis B, Davis FM, Harris OB, et al. Imaging the mammary gland and mammary tumours in 3D: optical tissue clearing and immunofluorescence methods. Breast Cancer Res 2016; 18(1):127. doi: 10.1186/ s13058-016-0754-9.
  38. Hu W, Tamadon A, Hsueh AJW, et al. Three-dimensional reconstruction of the vascular archi-tecture of the passive CLARITY-cleared mouse ovary. J Vis Exp 2017(130):56141. doi: 10.3791/56141.
  39. Zhang WL, Liu SH, Zhang WC, et al. Skeletal muscle CLARITY: A preliminary study of imaging the three-dimensional architecture of blood vessels and neurons. Cell J 2018; 20(2):132-137.
  40. Greenbaum A, Chan KY, Dobreva T, et al. Bone CLARITY: Clearing, imaging, and computational analysis of osteoprogenitors within intact bone marrow. Sci Transl Med 2017; 9(387): 6518. doi:10.1126/scitranslmed.aah6518.
  41. Singh JN, Nowlin TM, Seedorf GJ, et al. Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet microscopy. J Biomed Opt 2017; 22(7): 76011. doi: 10.1117/1.JBO.22.7.076011.
  42. Lisovsky A, Zhang DKY, Sefton MV. Effect of methacrylic acid beads on the sonic hedgehog signaling pathway and macrophage polarization in a subcutaneous injection mouse model. Biomaterials. 2016; 98:203-214.
  43. Khoradmehr A, Mazaheri F, Anvari M, et al. A simple technique for three-dimensional imaging and segmentation of brain vasculature using fast free-of-acrylamide clearing tissue in murine. Cell J 2019; 21(1):49-56.
Veterinary Research Forum
Volume 12, Issue 2
Spring 2021
Pages 167-173

  • Receive Date 03 December 2018
  • Revise Date 01 April 2019
  • Accept Date 17 April 2019

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