Vol 81, No 4 (2022)
Original article
Published online: 2021-11-05

open access

Page views 4449
Article views/downloads 628
Get Citation

Connect on Social Media

Connect on Social Media

Axonal quantification of the white matter association fasciculi in cerebral hemispheres of cow (Bos taurus), pig (Sus scrofa domesticus) and rabbit (Oryctolagus cuniculus)

M. Guerrero12, I. Romero23, C. Sandoval45, A. Gaibor-Pazmiño1, A. Noroña1, V. Zurita1, M. del Sol26, N. E. Ottone267
Pubmed: 34750803
Folia Morphol 2022;81(4):874-883.


Background: Cerebral white matter consists mainly of axons surrounded by myelin sheaths, which are grouped to form association, commissural, and projection fasciculi. The aim of our work was to quantify and compare under the microscope the axons of the white matter association fasciculi in the cerebral hemispheres of cow (Bos taurus), pig (Sus scrofa domesticus) and rabbit (Oryctolagus cuniculus) indirectly by identification of their myelin sheaths.
Materials and methods: The samples were taken from 30 cerebral hemispheres: 10 cow, 10 pig and 10 rabbit (15 right and 15 left). They were obtained following a protocol based on the Talairach-Tournoux coordinate system for human and primate brains. The slides were stained with Luxol Fast Blue, observed by optical microscopy, and photographed at 600×. Samples were also prepared for observation in scanning transmission electron microscopy with osmium tetroxide. The myelin sheaths/axons were counted with the ImageJ software.
Results: Statistically significant differences in the number of myelin sheaths per 410 μm2 were found in the inferior and superior longitudinal fasciculi between the left and right hemispheres of cows, with predominance of the right hemisphere; and in the inferior occipitofrontal fasciculus of the rabbit with predominance of the left hemisphere.
Conclusions: The use of animal models for experiments in the cerebral fasciculi, especially pig, could give us a greater understanding of the behaviour of demyelinating and neurodegenerative diseases in humans.

Article available in PDF format

View PDF Download PDF file


  1. Abdissa D, Hamba N, Gerbi A. Review Article on adult neurogenesis in humans. Transl Res Anat. 2020; 20: 100074.
  2. Adeeb N, Deep A, Hose N, et al. Stem cell therapy for spinal cord injury: The use of oligodendrocytes and motor neurons derived from human embryonic stem cells. Transl Res Anat. 2015; 1: 17–24.
  3. Andersen F, Watanabe H, Bjarkam C, et al. Pig brain stereotaxic standard space: mapping of cerebral blood flow normative values and effect of MPTP-lesioning. Brain Res Bull. 2005; 66(1): 17–29.
  4. Andronikou S. White Matter Tracts. In: (ed.). See Right Through Me. An Imaging Anatomy Atlas. Springer, Amsterdam 2012.
  5. Assaf Y, Pasternak O. Diffusion tensor imaging (DTI)-based white matter mapping in brain research: a review. J Mol Neurosci. 2008; 34(1): 51–61.
  6. Bürgel U, Amunts K, Hoemke L, et al. White matter fiber tracts of the human brain: three-dimensional mapping at microscopic resolution, topography and intersubject variability. Neuroimage. 2006; 29(4): 1092–1105.
  7. Carriel V, Campos A, Alaminos M, et al. Staining methods for normal and regenerative myelin in the nervous system. Methods Mol Biol. 2017; 1560: 207–218.
  8. Catani M, Thiebaut de Schotten M. A diffusion tensor imaging tractography atlas for virtual in vivo dissections. Cortex. 2008; 44(8): 1105–1132.
  9. Félix B, Léger ME, Albe-Fessard D, et al. Stereotaxic atlas of the pig brain. Brain Res Bull. 1999; 49(1-2): 1–137.
  10. Fernández-Miranda JC, Rhoton AL, Alvarez-Linera J, et al. Three-dimensional microsurgical and tractographic anatomy of the white matter of the human brain. Neurosurgery. 2008; 62(6 Suppl 3): 989–1026; discussion 1026.
  11. Guerrero M, del-Sol M, Ottone N. Preparación de Hemisferios Cerebrales para Disección de Tractos. Int J Morphol. 2019; 37(2): 533–540.
  12. Guerrero M, Veuthey C, Del Sol M, et al. Dissection of white matter association fasciculi in bovine (Bos taurus), pig (Sus scrofa domesticus) and rabbit (Oryctolagus cuniculus) brains. Anat Histol Embryol. 2020; 49(4): 550–562.
  13. Ikeda K, Shoin K, Mohri M, et al. Surgical indications and microsurgical anatomy of the transchoroidal fissure approach for lesions in and around the ambient cistern. Neurosurgery. 2002; 50(5): 1114–1120.
  14. Izrael M, Zhang P, Kaufman R, et al. Human oligodendrocytes derived from embryonic stem cells: Effect of noggin on phenotypic differentiation in vitro and on myelination in vivo. Mol Cell Neurosci. 2007; 34(3): 310–323.
  15. Keirstead HS, Nistor G, Bernal G, et al. Human embryonic stem cell-derived oligodendrocyte progenitor cell transplants remyelinate and restore locomotion after spinal cord injury. J Neurosci. 2005; 25(19): 4694–4705.
  16. Kiernan J. Histochemistry of staining methods for normal and degenerating myelin in the central and peripheral nervous systems. J Histotechnol. 2013; 30(2): 87–106.
  17. Liu XB, Schumann CM. Optimization of electron microscopy for human brains with long-term fixation and fixed-frozen sections. Acta Neuropathol Commun. 2014; 2: 42.
  18. Martino J, De Lucas EM. Subcortical anatomy of the lateral association fascicles of the brain: A review. Clin Anat. 2014; 27(4): 563–569.
  19. Melhem ER, Mori S, Mukundan G, et al. Diffusion tensor MR imaging of the brain and white matter tractography. Am J Roentgenol. 2002; 178(1): 3–16.
  20. Muñoz-Moreno E, Arbat-Plana A, Batalle D, et al. A magnetic resonance image based atlas of the rabbit brain for automatic parcellation. PLoS One. 2013; 8(7): e67418.
  21. Nistor GI, Totoiu MO, Haque N, et al. Human embryonic stem cells differentiate into oligodendrocytes in high purity and myelinate after spinal cord transplantation. Glia. 2005; 49(3): 385–396.
  22. Pascalau R, Szabo B. Fibre dissection and sectional study of the major porcine cerebral white matter tracts. Anat Histol Embryol. 2017; 46(4): 378–390.
  23. Paxinos J, Waltson C. The Rat Brain in Stereotaxic Coordinates. 5th ed. Elsevier Academic Press, Hoboken 2005.
  24. Pérez CJC, Gallegos SSP, Garduño PP, et al. Estandarización del método Klingler y su visualización tridimensional. Rev Hosp Jua Mex. 2008; 75(2): 99–108.
  25. Pistorio AL, Hendry SH, Wang X. A modified technique for high-resolution staining of myelin. J Neurosci Methods. 2006; 153(1): 135–146.
  26. Sawyer CH, Everett JW, Green JD. The rabbit diencephalon in stereotaxic coordinates. J Comp Neurol. 1954; 101(3): 801–824.
  27. Shah A, Jhawar SS, Goel A. Analysis of the anatomy of the Papez circuit and adjoining limbic system by fiber dissection techniques. J Clin Neurosci. 2012; 19(2): 289–298.
  28. Silva SM, Andrade JP. Neuroanatomy: The added value of the Klingler method. Ann Anat. 2016; 208: 187–193.
  29. Spiegel EA, Wycis HT, Freed H. Stereoencephalotomy in thalamotomy and related procedures. J Am Med Assoc. 1952; 148(6): 446–451.
  30. Talairach J, Tournoux P. Co-planar stereotaxic atlas of the human brain. 3-dimensional proportional system: an approach to cerebral imaging. Georg Thieme, New York 1998.
  31. Zemmoura I, Blanchard E, Raynal PI, et al. How Klingler's dissection permits exploration of brain structural connectivity? An electron microscopy study of human white matter. Brain Struct Funct. 2016; 221(5): 2477–2486.