Comparison of Commonly Used Retrograde Tracers in Rat Spinal Motor Neurons

Overview

Journal Neural Regen Res

Date 2015 Dec 23

PMID 26692873

Citations 5

Authors

You-Lai Yu

Hai-Yan Li

Pei-Xun Zhang

Xiao-Feng Yin

Na Han

Yu-Hui Kou

Bao-Guo Jiang

Affiliations

Soon will be listed here.

Abstract

The purpose of this study was to investigate the effect of four fluorescent dyes, True Blue (TB), Fluoro-Gold (FG), Fluoro-Ruby (FR), and 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), in retrograde tracing of rat spinal motor neurons. We transected the muscle branch of the rat femoral nerve and applied each tracer to the proximal stump in single labeling experiments, or combinations of tracers (FG-DiI and TB-DiI) in double labeling experiments. In the single labeling experiments, significantly fewer labeled motor neurons were observed after FR labeling than after TB, FG, or DiI, 3 days after tracer application. By 1 week, there were no significant differences in the number of labeled neurons between the four groups. In the double-labeling experiment, the number of double-labeled neurons in the FG-DiI group was not significantly different from that in the TB-DiI group 1 week after tracer application. Our findings indicate that TB, FG, and DiI have similar labeling efficacies in the retrograde labeling of spinal motor neurons in the rat femoral nerve when used alone. Furthermore, combinations of DiI and TB or FG are similarly effective. Therefore, of the dyes studied, TB, FG and DiI, and combinations of DiI with TB or FG, are the most suitable for retrograde labeling studies of motor neurons in the rat femoral nerve.

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References

Holmqvist B, Ostholm T, Ekstrom P . DiI tracing in combination with immunocytochemistry for analysis of connectivities and chemoarchitectonics of specific neural systems in a teleost, the Atlantic salmon. J Neurosci Methods. 1992; 42(1-2):45-63. DOI: 10.1016/0165-0270(92)90134-y. View

Madison R, Archibald S, Lacin R, Krarup C . Factors contributing to preferential motor reinnervation in the primate peripheral nervous system. J Neurosci. 1999; 19(24):11007-16. PMC: 6784932. View

Welin D, Novikova L, Wiberg M, Kellerth J, Novikov L . Survival and regeneration of cutaneous and muscular afferent neurons after peripheral nerve injury in adult rats. Exp Brain Res. 2007; 186(2):315-23. DOI: 10.1007/s00221-007-1232-5. View

Brushart T . Motor axons preferentially reinnervate motor pathways. J Neurosci. 1993; 13(6):2730-8. PMC: 6576505. View

Zhang L, McClellan A . Fluorescent tracers as potential candidates for double labeling of descending brain neurons in larval lamprey. J Neurosci Methods. 1999; 85(1):51-62. DOI: 10.1016/s0165-0270(98)00116-2. View

Zele T, Sketelj J, Bajrovic F . Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat. J Neurosci Methods. 2010; 191(2):208-14. DOI: 10.1016/j.jneumeth.2010.06.021. View

Maslany S, Crockett D, Egger M . Organization of cutaneous primary afferent fibers projecting to the dorsal horn in the rat: WGA-HRP versus B-HRP. Brain Res. 1992; 569(1):123-35. DOI: 10.1016/0006-8993(92)90378-m. View

Robinson G, Madison R . Motor neurons can preferentially reinnervate cutaneous pathways. Exp Neurol. 2004; 190(2):407-13. DOI: 10.1016/j.expneurol.2004.08.007. View

Vercelli A, Repici M, Garbossa D, Grimaldi A . Recent techniques for tracing pathways in the central nervous system of developing and adult mammals. Brain Res Bull. 2000; 51(1):11-28. DOI: 10.1016/s0361-9230(99)00229-4. View

10.

Katada A, Vos J, Swelstad B, Zealear D . A sequential double labeling technique for studying changes in motoneuronal projections to muscle following nerve injury and reinnervation. J Neurosci Methods. 2006; 155(1):20-7. DOI: 10.1016/j.jneumeth.2005.10.021. View

11.

Popratiloff A, Neiss W, Skouras E, Streppel M, Guntinas-Lichius O, Angelov D . Evaluation of muscle re-innervation employing pre- and post-axotomy injections of fluorescent retrograde tracers. Brain Res Bull. 2001; 54(1):115-23. DOI: 10.1016/s0361-9230(00)00411-1. View

12.

Brushart T . Preferential motor reinnervation: a sequential double-labeling study. Restor Neurol Neurosci. 2011; 1(3):281-7. DOI: 10.3233/RNN-1990-13416. View

13.

Harsh C, Archibald S, Madison R . Double-labeling of saphenous nerve neuron pools: a model for determining the accuracy of axon regeneration at the single neuron level. J Neurosci Methods. 1991; 39(2):123-30. DOI: 10.1016/0165-0270(91)90078-e. View

14.

Byers C, Fan R, Messina A, Morrison W, Galea M . Comparing the efficacy of two fluorescent retrograde tracers in labeling the motor and sensory neuron populations of the rat sciatic nerve. J Neurosci Methods. 2002; 114(2):159-64. DOI: 10.1016/s0165-0270(01)00520-9. View

15.

Gunturkun O, Melsbach G, HORSTER W, Daniel S . Different sets of afferents are demonstrated by the fluorescent tracers fast blue and rhodamine. J Neurosci Methods. 1993; 49(1-2):103-11. DOI: 10.1016/0165-0270(93)90114-7. View

16.

Richmond F, Gladdy R, Creasy J, Kitamura S, Smits E, Thomson D . Efficacy of seven retrograde tracers, compared in multiple-labelling studies of feline motoneurones. J Neurosci Methods. 1994; 53(1):35-46. DOI: 10.1016/0165-0270(94)90142-2. View

17.

Novikova L, Novikov L, Kellerth J . Persistent neuronal labeling by retrograde fluorescent tracers: a comparison between Fast Blue, Fluoro-Gold and various dextran conjugates. J Neurosci Methods. 1997; 74(1):9-15. DOI: 10.1016/s0165-0270(97)02227-9. View

18.

Robinson G, Madison R . Preferential motor reinnervation in the mouse: comparison of femoral nerve repair using a fibrin sealant or suture. Muscle Nerve. 2003; 28(2):227-31. DOI: 10.1002/mus.10422. View

19.

Hayashi A, Moradzadeh A, Hunter D, Kawamura D, Puppala V, Tung T . Retrograde labeling in peripheral nerve research: it is not all black and white. J Reconstr Microsurg. 2007; 23(7):381-9. DOI: 10.1055/s-2007-992344. View

20.

Deng C, Rogers L . Differential sensitivities of the two visual pathways of the chick to labelling by fluorescent retrograde tracers. J Neurosci Methods. 1999; 89(1):75-86. DOI: 10.1016/s0165-0270(99)00044-8. View