A neuroanatomical study of the feline brain using MRI and mulligan staining: functional and pathological considerations

Document Type : Full paper (Original article)


1 Department of Anatomy and Compared Pathological Anatomy, Veterinary Faculty, Campus de Espinardo, University of Murcia, 30100, Murcia, Spain

2 Department of Morphology, Veterinary Faculty, University of Las Palmas de Gran Canaria, Trasmontaña, Arucas, 35413, Las Palmas, Spain

3 Department of Medicine and Surgery, Veterinary Faculty, Campus de Espinardo, University of Murcia, 30100, Murcia, Spain

4 Division of Veterinary Science Centre, University College Dublin, School of Veterinary Medicine, University of Dublin, Belfield, Dublin 4, Ireland


Background: Despite multiple studies describing accurate diagnoses using advanced neuroimaging techniques, low and mid-field magnetic resonance imaging (MRI) are still the most frequent scanners in veterinary clinics. To date, these studies in cats do not show a clear distinction of nerve centres in MRI data. Aims: The objective of this study is to determine the efficacy of Mulligan histological staining as a tool in facilitating the location and identification of the main structures of the feline brain in MRI. This study aims to facilitate the interpretation of MRI obtained with these types of scanners. Methods: A total of 10 feline brains were used. One specimen was used for MRI (T2 sequence using a 1.5T scanner). The other 9 brains were sectioned and stained with the three Mulligan staining techniques (Mulligan, Le Masurier and Robert). Results: The uptake of stain by the grey matter in these sections allowed the determination of the location and the limits of these nervous structures within the brain. The histological location of these structures was correlated with the MRI scans, leading to the successful identification of many small, indistinct nuclei. Conclusion: Mulligan staining is proposed as a tool that facilitates the location of nerve structures in comparison with data from the most frequently-used MRI scanners in veterinary clinics.


Main Subjects

Barnard, JW; Roberts, JO and Brown, JG (1949). A simple macroscopic staining and mounting procedure for wet sections from cadaver brains. Anat. Rec., 105: 11-17.
Claßen, AC; Kneissl, S; Lang, J; Tichy, A and Pakazdy, A (2016). Magnetic resonance features of the feline hippocampus in epileptic and non-epileptic cats: a blinded, retrospective, multi-observer study. BMC Vet. Res., 12: 165-171.
Crick, FC and Koch, C (2005). What is the function of the claustrum? Philos. Trans. R. Soc. Lond. B Biol. Sci., 360: 1271-1279.
Das, A and Takahashi, E (2018). Characterization of white matter tracts by diffusion MR tractography in cat and ferret that have similar gyral patterns. Cereb. Cortex., 28: 1338-1347.
De Meneses, MS; Montano, JCP; Fuzza, RF and Milano, JB (2004). Comparative analysis of sections of human brain stained by three different techniques. Arq. Neuropsiquiatr., 62: 276-281.
Dos Santos, EC; Da Luz Veronez, DA; De Almeida, DB; Piedade, GS; Oldoni, C; De Meneses, MS and Marques, MS (2019). Morphometric study of the internal globus pallidus using the Robert, Barnard, and Brown staining method. World Neurosurg., 126: e371-e378.
Ghaznawi, R; De Bresser, J; Van der Graaf, Y; Zwartbol, MH; Witkamp, TD; Geerlings, MI; Hendrikse, J and SMART Study Group (2018). Detection and characterization of small infarcts in the caudate nucleus on 7 Tesla MRI: The SMART-MR study. J. Cereb. Blood Flow Metab., 38: 1609-1617.
Gray-Edwards, HL; Salibi, N; Josephson, EM; Hudson, JA; Cox, NR; Randle, AN; McCurdy, VJ; Bradbury, AM; Wilson, DU; Beyers, RJ; Denney, TS and Martin, DR (2014). High resolution MRI anatomy of the cat brain at 3 Tesla. J. Neurosci. Methods. 227: 10-17.
Hespel, AM and Cole, RC (2018). Advances in high-field MRI. Vet. Clin. North Am. Small Anim. Pract., 48: 11-21.
Hudson, LC; Cauzinille, L; Kornegay, JN and Tompkins, MB (1995). Magnetic resonance imaging of the normal feline brain. Vet. Radiol. Ultrasound., 36: 267-275.
Johnson, PJ; Pascalau, R; Luh, WM; Raj, A; Cerda-Gonzalez, S and Barry, EF (2020). Stereotaxic diffusion tensor imaging white matter atlas for the in vivo domestic feline brain. Front. Neuroanat., 14: 1-13.
Leigh, EJ; Mackillop, E; Robertson, ID and Hudson, LC (2008). Clinical anatomy of the canine brain using magnetic resonance imaging. Vet. Radiol. Ultrasound., 49: 113-121.
LeMasurier, HE (1935). Simple method of staining macroscopic brain sections. Arq. Neuropsych., 34: 1065-1067.
Mogicato, G; Conchou, F; Layssol-Lamour, C; Raharison, F and Sautet, J (2012). Normal feline brain: clinical anatomy using magnetic resonance imaging. Anat. Histol. Embryol., 41: 87-95.
Mooncey, MS and Sagoo, MG (2014). Comparative staining methods with room temperature plastination (15-18ºC) of brain specimens,using Biodur S10/S3. J. Plast., 26: 21-29.
Müllhaupt, D; Augsburger, H; Schwarz, A; Fischer, G; Kircher, P; Hatt, JM and Ohlerth, S (2015). Magnetic resonance imaging anatomy of the rabbit brain at 3 T. Acta Vet. Scand., 57: 47-55.
Mulligan, JH (1931). A method of staining the brain for macroscopic study. J. Anat., 65: 468-472.
Nakamura, H (2018). Cerebellar projections to the ventral lateral geniculate nucleus and the thalamic reticular nucleus in the cat. J. Neurosci. Res., 96: 63-74.
Nickel, R; Schummer, A and Seiferle, E (1988). Anatomical treatise of domestic mammals. Vol. IV, 1st Edn., Casa Editrice Ambrosiana. Milano. PP: 1-439.
Oldoni, C; Luz Veronez, DAD; Piedade, GS; Santos, ECD; Almeida, DB and Meneses, MS (2018). Morphometric analysis of the nucleus accumbens using the Mulligan staining method. World Neurosurg., 118: e223-e228.
Onodera, S and Hicks, TP (2009). A comparative neuroanatomical study of the red nucleus of the cat,
macaque and human. PloS One. 4: e6623-e6641.
Park, J (2016). Movement disorders following cerebrovascular lesion in the basal ganglia circuit. J. Mov. Disord., 9: 71-79.
Przyborowska, P; Adamiak, Z; Holak, P and Zhalniarovich, Y (2017). Comparison of feline brain anatomy in 0.25 and 3 Tesla magnetic resonance images. Anat. Histol. Embryol., 46: 178-186.
Sandoval, J (2000). Veterinary anatomy treatise. Vol. III: Head and visceral systems. 2nd Edn., Leon: Imprenta Sorles. PP: 1-457.
Stolzberg, D; Wong, C; Butler, BE and Lomber, SG (2017). Catlas: An magnetic resonance imaging-based three-dimensional cortical atlas and tissue probability maps for the domestic cat (Felis catus). J. Comp. Neurol., 525: 3190-3206.
Sugiyama, A; Sato, N; Kimura, Y; Ota, M; Maekawa, T; Sone, D; Enokizono, M; Murata, M; Matsuda, H and Kuwabara, S (2018). MR findings in the substantia nigra on phase difference enhanced imaging in neuro-degenerative parkinsonism. Parkinsonism Relat Disord., 48: 10-16.
Villablanca, JR (2010). Why do we have a caudate nucleus? Acta Neurobiol. Exp. (Wars)., 70: 95-105.
Zhang, XY; Wang, JJ and Zhu, JN (2016). Cerebellar fastigial nucleus: from anatomic construction to physiological functions. Cerebellum Ataxias., 3: 9-18.