Abstract
The fluoride resistant acid phosphatase (FRAP) is considered a biomarker enzyme of presence and function in nociceptive fibers. Its well described spinal dorsal horn (DH) superficial lamina location contrasts to its less known cervico-lumbar DH distribution. Thus, 15 male Sprague Dawley rats (300–400 g) were anaesthetized (thiopental 60 mg kg−1 i.p.) and laminectomized (C 1-L 5) to extract and fix (formaline 4% + NaF 1%) spinal cords. Cervical, thoracic and lumbar segments were coronally cut (1 mm slices) and micropunched (1 mmØ) at the dorsal horn (both left and right). Samples were diluted in NaF 1% in saline (1000 μL: fragment) and homogenized. Protein concentration of the homogenate was determined by Bradford method to correct variation on tissue sampling and used to compensate enzymatic dilution. FRAP activity was determined by the Gomorri method and expressed as U gr prot−1 L−1. No differences were found between left and right DH FRAP activities (Mann-Whitney U Test, P > 0.05) in any metamera. DH segment values were: cervical 2.53 (2.67–2.40 U gr prot−1 L−1; mean ± 95% CI), thoracic 3.59 (3.96–3.21) and lumbar 2.42 (2.75–2.09). Thoracic FRAP activity was statistically higher (+45.3%) than those from cervical and lumbar DH segments (ANOVA; Ω2 = 0.29; F = 17.78; P = 0.000001; Levene test P = 0.000006). No significant correlation was found among cervicotoraco-lumbar FRAP activity. The increased FRAP activity for thoracic DH could be explained by the thoracic overlapped primary afferents and collateral fibers with cervical or lumbar input added to the basal thoracic FRAP expression.
References
Bosco, R., Alvarado, S., Quiroz, Daniel, and Eblen-Zajjur, A., Digital cytomorphometry of neurons from de dorsal root ganglion of the rat, J. Histotechnol., 2010, vol. 33, pp. 113–118.
Knyihar-Csillik, E., Bezzegh, A., Boti, S., and Csillik, B., Thiamine monophosphatase: a genuine marker for transganglionic regulation of primary sensory neurons, J. Histochem. Cytochem., 1986, vol. 34, pp. 363–371.
Nagy, J.I. and Hunt, S.P. Fluoride-resistant acid phosphatase-containing neurones in dorsal root ganglia are separate from those containing substance P or somatostatin, Neuroscience, 1982, vol. 7, pp. 89–97.
Sanyal, S. and Rustioni, A., Phosphatases in the substantia gelatinosa and motoneurones: a comparative histochemical study, Brain Res., 1914, vol. 76, pp. 161–166.
Taylor-Blake, B. and Zylka, M.J., Prostatic acid phosphatase is expressed in peptidergic and nonpeptidergic nociceptive neurons of mice and rats, PLoS ONE., 2010, vol. 5, p. e8674.
Jansco, G. and Knyihar, E., Functional linkage between nociception and fluoride-resistant acid phosphatase activity in the Rolando substance, Neurobiology, 1975, vol. 5, pp. 42–43.
Knyihar, E. and Csillik, B., FRAP: Histochemistry of the primary nociceptive neuron, Progr. Histochem. Cytochem., 1981, vol. 14, pp. 1–137.
Glykys, J., Guadama, M., Marcano, L., Ochoa, E., and Eblen-Zajjur, A., Inflammation induced increase of fluoride resistant acid phosphatase (FRAP) activity in the spinal dorsal horn in rats, Neurosci. Letters., 2003, vol. 337, pp. 167–169.
Kantner, R. and Kirby, M., Changes in acid phosphatase activity in the substantia gelatinosa in response to pain, Brain Res., 1982, vol. 238, pp. 451–456.
Silverman, J. and Kruger, L., Acid phosphatase as a selective marker for a class of small sensory ganglion cells in several mammals: spinal cord distribution, histochemical properties, and relation to fluoride-resistant acid phosphatase (FRAP) of rodents, Somatosens. Res., 1988, vol. 5, pp. 219–246.
Devor, M. and Claman, D., Mapping and plasticity of acid phosphatase afferents in the rat dorsal horn, Brain Res., 1990, vol. 190, pp. 17–28.
Fieschi, C. and Soriani, S., Enzymatic activities in the spinal cord after scitic section, alkaline and acid phosphatases, 5-nucleotidase and ATP-ase, J. Neurochemistry, 1959, vol. 4, pp. 71–77.
Willis, W. and Coggeshall, R., Sensory Mechanisms of the Spinal Cord: Primary Afferent Neurons and the Spinal Dorsal Horn, New York: Plenum Press, 2004, vol. 1, 3rd ed.
Brown, A.G. and Iggo, A., A quantitative study of cutaneous receptors and afferent fibres in the cat and rabbit, J. Physiol., 1967, vol. 193, pp. 707–733.
Brown, A.G., The dorsal horn of the spinal cord, Quart. J. Exp. Physiol., 1982, vol. 67, pp. 193–212.
Réthelyi, M., Trevino, D.L., and Perl, E.R., Distribution of primary afferent fibers within the sacrococcygeal dorsal horn: an autoradiographic study, J. Comp. Neurol., 1979, vol. 185, pp. 603–621.
Wall, P.D. and Werman, R., The physiology and anatomy of long-ranging afferent fibres within the spinal cord, J. Physiol., 1976, vol. 255, pp. 321–334.
Lamotte, C.C., Kapadia, S.E., and Shapiro, Ch.M., Central projections of the sciatic, saphenous, median, and ulnar nerves of the rat demonstrated by transganglionic transport of choleragenoid-HRP (B-HRP) and wheat germ agglutinin-HRP (WGA-HRP), J. Comp. Neurol., 1991, vol. 311, pp. 546–562.
Swett, J.E. and Woolf, C.J., The somatotopic organization of primary afferent terminals in the superficial laminae of the dorsal horn of the rat spinal cord, J. Comp. Neurol., 1985, vol. 231, pp. 66–77.
Wilson, P. and Kitchener, P.D., Plasticity of cutaneous primary afferent projections to the spinal dorsal horn, Prog. Neurobiol., 1996, vol. 48, pp. 105–129.
Wall, P.D. and Wolstencroft, J.H., The presence of ineffective synapses and the circumstances which unmask them, Phil. Trans. R. Soc. Lond. B, 1977, vol. 278, pp. 361–372.
Cadden, S.W., Villanueva, L., Chitour, D., and Le Bars, D., Depression of activities of dorsal horn convergent neurones by propriospinal mechanisms triggered by noxious inputs; comparison with diffuse noxious inhibitory controls (DNIC), Brain Res., 1983, vol. 275, pp. 1–11.
Sandkühler, J., Stelzer, B., and Fu, Q.-F., Characteristics of propriospinal modulation of nociceptive lumbar spinal dorsal horn neurons in the cat, Neuroscience, 1993, vol. 54, pp. 957–967.
Author information
Authors and Affiliations
Corresponding author
Additional information
The article is published in the original.
Rights and permissions
About this article
Cite this article
Sosa, D., Velasco, D., Valenzuela, S. et al. Mapping of the fluoride resistant acid phosphatase (FRAP) activity in cervico-thoracic-lumbar spinal dorsal horn in rats. Neurochem. J. 7, 313–315 (2013). https://doi.org/10.1134/S1819712413040119
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1819712413040119