Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents

MM Mesulam

Tetramethyl benzidine (TMB) is a presumptively non-carcinogenic chromogen which yields a blue reaction-product at sites of horseradish peroxidase activity. Sixty-six distinct procedures were performed in rats and monkeys in order to determine the optimal incubation parameters for TMB. As a result, a procedure is recommended whose sensitivity greatly surpasses that of a previously described benzidine dihydrochloride method. Indeed, the sensitivity of this new method in demonstrating retrograde transport is markedly superior to that of the previously described benzidine dihydrochloride method. Furthermore, as a consequence of this enhanced sensitivity, many efferent connections of the injection site are also visualized. The injection site demonstrated by this TMB procedure is significantly larger than the one demonstrated when benzidine dihydrochloride or diaminobenzidine is used as a chromogen. Finally, this TMB procedure has been compared to two other TMB procedures and found to provide superior morphology and sensitivity.

Volume 26, Issue 2, pp. 106-117, 02/01/1978
Copyright © 1978 by The Histochemical Society

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				H Liu, N Schuelert, J J McDougall, and S S Lee Central neural activation of hyperdynamic circulation in portal hypertensive rats depends on vagal afferent nerves Gut, July 1, 2008; 57(7): 966 - 973. [Abstract] [Full Text] [PDF]

				R. D. Lane, C. P. Pluto, C. L. Kenmuir, N. L. Chiaia, and R. D. Mooney Does Reorganization in the Cuneate Nucleus Following Neonatal Forelimb Amputation Influence Development of Anomalous Circuits Within the Somatosensory Cortex? J Neurophysiol, February 1, 2008; 99(2): 866 - 875. [Abstract] [Full Text] [PDF]

				H. Mizuno, T. Hirano, and Y. Tagawa Evidence for Activity-Dependent Cortical Wiring: Formation of Interhemispheric Connections in Neonatal Mouse Visual Cortex Requires Projection Neuron Activity J. Neurosci., June 20, 2007; 27(25): 6760 - 6770. [Abstract] [Full Text] [PDF]

				Y. Izawa, Y. Sugiuchi, and Y. Shinoda Neural Organization of the Pathways From the Superior Colliculus to Trochlear Motoneurons J Neurophysiol, May 1, 2007; 97(5): 3696 - 3712. [Abstract] [Full Text] [PDF]

				H. Ishii, T. Niioka, E. Sudo, and H. Izumi Evidence for parasympathetic vasodilator fibres in the rat masseter muscle J. Physiol., December 1, 2005; 569(2): 617 - 629. [Abstract] [Full Text] [PDF]

				T. L. Powley, M. M. Chi, E. A. Baronowsky, and R. J. Phillips Gastrointestinal tract innervation of the mouse: afferent regeneration and meal patterning after vagotomy Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2005; 289(2): R563 - R574. [Abstract] [Full Text] [PDF]

				J. A. Brzezinski IV, N. L. Brown, A. Tanikawa, R. A. Bush, P. A. Sieving, M. H. Vitaterna, J. S. Takahashi, and T. Glaser Loss of Circadian Photoentrainment and Abnormal Retinal Electrophysiology in Math5 Mutant Mice Invest. Ophthalmol. Vis. Sci., July 1, 2005; 46(7): 2540 - 2551. [Abstract] [Full Text] [PDF]

				J. T. Green and J. E. Steinmetz Purkinje cell activity in the cerebellar anterior lobe after rabbit eyeblink conditioning Learn. Mem., May 1, 2005; 12(3): 260 - 269. [Abstract] [Full Text] [PDF]

				R. Matsumoto, D. R. Nair, E. LaPresto, I. Najm, W. Bingaman, H. Shibasaki, and H. O. Luders Functional connectivity in the human language system: a cortico-cortical evoked potential study Brain, October 1, 2004; 127(10): 2316 - 2330. [Abstract] [Full Text] [PDF]

				C. Francois, D. Grabli, K. McCairn, C. Jan, C. Karachi, E.-C. Hirsch, J. Feger, and L. Tremblay Behavioural disorders induced by external globus pallidus dysfunction in primates II. Anatomical study Brain, September 1, 2004; 127(9): 2055 - 2070. [Abstract] [Full Text] [PDF]

				L. R. Dehner, L. P. Keniston, H. R. Clemo, and M. A. Meredith Cross-modal Circuitry Between Auditory and Somatosensory Areas of the Cat Anterior Ectosylvian Sulcal Cortex: A 'New' Inhibitory Form of Multisensory Convergence Cereb Cortex, April 1, 2004; 14(4): 387 - 403. [Abstract] [Full Text] [PDF]

				L. Cao, L. Liu, Z.-Y. Chen, L.-M. Wang, J.-L. Ye, H.-Y. Qiu, C.-L. Lu, and C. He Olfactory ensheathing cells genetically modified to secrete GDNF to promote spinal cord repair Brain, March 1, 2004; 127(3): 535 - 549. [Abstract] [Full Text] [PDF]

				X.-Y. Song, J.-h. Zhong, X. Wang, and X.-F. Zhou Suppression of p75NTR Does Not Promote Regeneration of Injured Spinal Cord in Mice J. Neurosci., January 14, 2004; 24(2): 542 - 546. [Abstract] [Full Text] [PDF]

				K. McBride, B. Slotnick, and F. L. Margolis Does Intranasal Application of Zinc Sulfate Produce Anosmia in the Mouse? An Olfactometric and Anatomical Study Chem Senses, October 1, 2003; 28(8): 659 - 670. [Abstract] [Full Text] [PDF]

				S. Takeda, M. Kondo, J. Sasaki, H. Kurahashi, H. Kano, K. Arai, K. Misaki, T. Fukui, K. Kobayashi, M. Tachikawa, et al. Fukutin is required for maintenance of muscle integrity, cortical histiogenesis and normal eye development Hum. Mol. Genet., June 15, 2003; 12(12): 1449 - 1459. [Abstract] [Full Text] [PDF]

				H.-L. Pan, G. M. Khan, K. D. Alloway, and S.-R. Chen Resiniferatoxin Induces Paradoxical Changes in Thermal and Mechanical Sensitivities in Rats: Mechanism of Action J. Neurosci., April 1, 2003; 23(7): 2911 - 2919. [Abstract] [Full Text] [PDF]

				A. Gruart, M. Streppel, O. Guntinas-Lichius, D. N. Angelov, W. F. Neiss, and J. M. Delgado-Garcia Motoneuron adaptability to new motor tasks following two types of facial-facial anastomosis in cats Brain, January 1, 2003; 126(1): 115 - 133. [Abstract] [Full Text] [PDF]

				C. L. Jordan, S. E. Christensen, R. J. Handa, J. L. Anderson, W. A. Pouliot, and S. M. Breedlove Evidence That Androgen Acts Through NMDA Receptors to Affect Motoneurons in the Rat Spinal Nucleus of the Bulbocavernosus J. Neurosci., November 1, 2002; 22(21): 9567 - 9572. [Abstract] [Full Text] [PDF]

				G. Orieux, C. Francois, J. Feger, and E. C. Hirsch Consequences of Dopaminergic Denervation on the Metabolic Activity of the Cortical Neurons Projecting to the Subthalamic Nucleus in the Rat J. Neurosci., October 1, 2002; 22(19): 8762 - 8770. [Abstract] [Full Text] [PDF]

				K. Mousavi, W. Miranda, and D. J. Parry Neurotrophic factors enhance the survival of muscle fibers in EDL, but not SOL, after neonatal nerve injury Am J Physiol Cell Physiol, September 1, 2002; 283(3): C950 - C959. [Abstract] [Full Text] [PDF]

				Y. Shostak, Y. Ding, J. Mavity-Hudson, and V. A. Casagrande Cortical Synaptic Arrangements of the Third Visual Pathway in Three Primate Species: Macaca mulatta, Saimiri sciureus, and Aotus trivirgatus J. Neurosci., April 1, 2002; 22(7): 2885 - 2893. [Abstract] [Full Text] [PDF]

				T. C. Chou, A. A. Bjorkum, S. E. Gaus, J. Lu, T. E. Scammell, and C. B. Saper Afferents to the Ventrolateral Preoptic Nucleus J. Neurosci., February 1, 2002; 22(3): 977 - 990. [Abstract] [Full Text] [PDF]

				Y.-J. Yan, D.-M. Cui, and J. C. Lynch Overlap of Saccadic and Pursuit Eye Movement Systems in the Brain Stem Reticular Formation J Neurophysiol, December 1, 2001; 86(6): 3056 - 3060. [Abstract] [Full Text] [PDF]

				E. A. Fox, R. J. Phillips, E. A. Baronowsky, M. S. Byerly, S. Jones, and T. L. Powley Neurotrophin-4 Deficient Mice Have a Loss of Vagal Intraganglionic Mechanoreceptors from the Small Intestine and a Disruption of Short-Term Satiety J. Neurosci., November 1, 2001; 21(21): 8602 - 8615. [Abstract] [Full Text] [PDF]

				E. C. Tsai, R. L. van Bendegem, S. W. Hwang, and C. H. Tator A Novel Method for Simultaneous Anterograde and Retrograde Labeling of Spinal Cord Motor Tracts in the Same Animal J. Histochem. Cytochem., September 1, 2001; 49(9): 1111 - 1122. [Abstract] [Full Text] [PDF]

				K. J. Huffman and L. Krubitzer Area 3a: Topographic Organization and Cortical Connections in Marmoset Monkeys Cereb Cortex, September 1, 2001; 11(9): 849 - 867. [Abstract] [Full Text] [PDF]

				B. P. Kolomiets, J. M. Deniau, P. Mailly, A. Menetrey, J. Glowinski, and A. M. Thierry Segregation and Convergence of Information Flow through the Cortico-Subthalamic Pathways J. Neurosci., August 1, 2001; 21(15): 5764 - 5772. [Abstract] [Full Text] [PDF]

				S. Millecamps, D. Nicolle, I. Ceballos-Picot, J. Mallet, and M. Barkats Synaptic sprouting increases the uptake capacities of motoneurons in amyotrophic lateral sclerosis mice PNAS, June 7, 2001; (2001) 131031098. [Abstract] [Full Text] [PDF]

				H. Komatsu, M. Kinoshita, and I. Murakami Neural Responses in the Retinotopic Representation of the Blind Spot in the Macaque V1 to Stimuli for Perceptual Filling-In J. Neurosci., December 15, 2000; 20(24): 9310 - 9319. [Abstract] [Full Text] [PDF]

				D. O. Frost, D. Boire, G. Gingras, and M. Ptito Surgically created neural pathways mediate visual pattern discrimination PNAS, September 19, 2000; (2000) 190179997. [Abstract] [Full Text]

				M. A. Ruda, Q.-D. Ling, A. G. Hohmann, Y. B. Peng, and T. Tachibana Altered Nociceptive Neuronal Circuits After Neonatal Peripheral Inflammation Science, July 28, 2000; 289(5479): 628 - 630. [Abstract] [Full Text]

				F. Clasca, A. Llamas, and F. Reinoso-Suarez Cortical Connections of the Insular and Adjacent Parieto-temporal Fields in the Cat Cereb Cortex, April 1, 2000; 10(4): 371 - 399. [Abstract] [Full Text] [PDF]

				B. Conway, J. D. Boyd, T. H. Stewart, and J. A. Matsubara The Projection from V1 to Extrastriate Area 21a: A Second Patchy Efferent Pathway that Colocalizes with the CO Blob Columns in Cat Visual Cortex Cereb Cortex, February 1, 2000; 10(2): 149 - 159. [Abstract] [Full Text] [PDF]

				C. Dean, L. K. Geiger, B. M. Sprtel, P. J. Ohtake, and H. V. Forster An anatomic atlas of the medulla oblongata of the adult goat J Appl Physiol, September 1, 1999; 87(3): 1220 - 1229. [Abstract] [Full Text] [PDF]

				L. E. White, W. H. Bosking, S. M. Williams, and D. Fitzpatrick Maps of Central Visual Space in Ferret V1 and V2 Lack Matching Inputs from the Two Eyes J. Neurosci., August 15, 1999; 19(16): 7089 - 7099. [Abstract] [Full Text] [PDF]

				P. G. Sullivan, A. J. Bruce-Keller, A. G. Rabchevsky, S. Christakos, D. K. St. Clair, M. P. Mattson, and S. W. Scheff Exacerbation of Damage and Altered NF-kappa B Activation in Mice Lacking Tumor Necrosis Factor Receptors after Traumatic Brain Injury J. Neurosci., August 1, 1999; 19(15): 6248 - 6256. [Abstract] [Full Text] [PDF]

				Y. Izawa, Y. Sugiuchi, and Y. Shinoda Neural Organization From the Superior Colliculus to Motoneurons in the Horizontal Oculomotor System of the Cat J Neurophysiol, June 1, 1999; 81(6): 2597 - 2611. [Abstract] [Full Text] [PDF]

				M. I. Romero, M. A. Romero, and G. M. Smith Visualization of Axonally Transported Horseradish Peroxidase Using Enhanced Immunocytochemical Detection: A Direct Comparison with the Tetramethylbenzidine Method J. Histochem. Cytochem., February 1, 1999; 47(2): 265 - 272. [Abstract] [Full Text]

				E. Brustein and S. Rossignol Recovery of Locomotion After Ventral and Ventrolateral Spinal Lesions in the Cat. I. Deficits and Adaptive Mechanisms J Neurophysiol, September 1, 1998; 80(3): 1245 - 1267. [Abstract] [Full Text] [PDF]

				R. J. Phillips and T. L. Powley Gastric volume detection after selective vagotomies in rats Am J Physiol Regulatory Integrative Comp Physiol, June 1, 1998; 274(6): R1626 - R1638. [Abstract] [Full Text] [PDF]

				A. Ramon-Cueto, G. W. Plant, J. Avila, and M. B. Bunge Long-Distance Axonal Regeneration in the Transected Adult Rat Spinal Cord Is Promoted by Olfactory Ensheathing Glia Transplants J. Neurosci., May 15, 1998; 18(10): 3803 - 3815. [Abstract] [Full Text] [PDF]

				M. Kressel Tyramide Amplification Allows Anterograde Tracing by Horseradish Peroxidase-conjugated Lectins in Conjunction with Simultaneous Immunohistochemistry J. Histochem. Cytochem., April 1, 1998; 46(4): 527 - 534. [Abstract] [Full Text] [PDF]

				N. G. Forger, M. L. Howell, L. Bengston, L. MacKenzie, T. M. DeChiara, and G. D. Yancopoulos Sexual Dimorphism in the Spinal Cord Is Absent in Mice Lacking the Ciliary Neurotrophic Factor Receptor J. Neurosci., December 15, 1997; 17(24): 9605 - 9612. [Abstract] [Full Text] [PDF]

				L. Proctor and M. Konishi Representation of sound localization cues in the auditory thalamus of the barn owl PNAS, September 16, 1997; 94(19): 10421 - 10425. [Abstract] [Full Text] [PDF]

				B. Brett and D. S. Barth Subcortical Modulation of High-Frequency (Gamma Band) Oscillating Potentials in Auditory Cortex J Neurophysiol, August 1, 1997; 78(2): 573 - 581. [Abstract] [Full Text] [PDF]

				R. Grill, K. Murai, A. Blesch, F. H. Gage, and M. H. Tuszynski Cellular Delivery of Neurotrophin-3 Promotes Corticospinal Axonal Growth and Partial Functional Recovery after Spinal Cord Injury J. Neurosci., July 15, 1997; 17(14): 5560 - 5572. [Abstract] [Full Text] [PDF]

				J. S. Kanwal and T. E. Finger Parallel Medullary Gustatospinal Pathways In a Catfish: Possible Neural Substrates for Taste-Mediated Food Search J. Neurosci., June 15, 1997; 17(12): 4873 - 4885. [Abstract] [Full Text] [PDF]

				D. J. Felleman, Y. Xiao, and E. McClendon Modular Organization of Occipito-Temporal Pathways: Cortical Connections between Visual Area 4 and Visual Area 2 and Posterior Inferotemporal Ventral Area in Macaque Monkeys J. Neurosci., May 1, 1997; 17(9): 3185 - 3200. [Abstract] [Full Text] [PDF]

				H.B. Parthasarathy and A.M. Graybiel Cortically Driven Immediate-Early Gene Expression Reflects Modular Influence of Sensorimotor Cortex on Identified Striatal Neurons in the Squirrel Monkey J. Neurosci., April 1, 1997; 17(7): 2477 - 2491. [Abstract] [Full Text] [PDF]

				A. Angelucci, F. Clasca, E. Bricolo, K. S. Cramer, and M. Sur Experimentally Induced Retinal Projections to the Ferret Auditory Thalamus: Development of Clustered Eye-Specific Patterns in a Novel Target J. Neurosci., March 15, 1997; 17(6): 2040 - 2055. [Abstract] [Full Text] [PDF]

				K. S. Cramer, A. Angelucci, J.-O. Hahm, M. B. Bogdanov, and M. Sur A Role for Nitric Oxide in the Development of the Ferret Retinogeniculate Projection J. Neurosci., December 15, 1996; 16(24): 7995 - 8004. [Abstract] [Full Text] [PDF]

				F. C. Zhou, Y. H. Chiang, and Y. Wang Constructing a New Nigrostriatal Pathway in the Parkinsonian Model with Bridged Neural Transplantation in Substantia Nigra J. Neurosci., November 1, 1996; 16(21): 6965 - 6974. [Abstract] [Full Text] [PDF]

				J. C. Horton and D. R. Hocking Anatomical Demonstration of Ocular Dominance Columns in Striate Cortex of the Squirrel Monkey J. Neurosci., September 1, 1996; 16(17): 5510 - 5522. [Abstract] [Full Text] [PDF]

				R. J. Mannion, T. P. Doubell, R. E. Coggeshall, and C. J. Woolf Collateral Sprouting of Uninjured Primary Afferent A-Fibers into the Superficial Dorsal Horn of the Adult Rat Spinal Cord after Topical Capsaicin Treatment to the Sciatic Nerve J. Neurosci., August 15, 1996; 16(16): 5189 - 5195. [Abstract] [Full Text] [PDF]

				M. Curfs, A. Gribnau, and P. Dederen Postnatal maturation of the dendritic fields of motoneuron pools supplying flexor and extensor muscles of the distal forelimb in the rat Development, January 2, 1993; 117(2): 535 - 541. [Abstract] [PDF]

				M Sur, P. Garraghty, and A. Roe Experimentally induced visual projections into auditory thalamus and cortex Science, December 9, 1988; 242(4884): 1437 - 1441. [Abstract] [PDF]

				G Aston-Jones, M Ennis, V. Pieribone, W. Nickell, and M. Shipley The brain nucleus locus coeruleus: restricted afferent control of a broad efferent network Science, November 7, 1986; 234(4777): 734 - 737. [Abstract] [PDF]

				M. R. BRAFORD JR. De Gustibus Non Est Disputandem: A Spiral Center for Taste in the Brain of the Teleost Fish, Heterotis niloticus Science, April 25, 1986; 232(4749): 489 - 491. [Abstract] [PDF]

				E. Kurz, D. Sengelaub, and A. Arnold Androgens regulate the dendritic length of mammalian motoneurons in adulthood Science, April 18, 1986; 232(4748): 395 - 398. [Abstract] [PDF]