Skip to main content

Diamidino yellow dihydrochloride (DY·2HCl); a new fluorescent retrograde neuronal tracer, which migrates only very slowly out of the cell

Summary

Earlier studies showed that Nuclear Yellow (NY), True Blue (TB) and Fast Blue (FB) are transported retrogradely through axons to their parent cell bodies. NY produces a yellow fluorescent labeling of the neuronal nucleus at 360 nm excitation wavelength, while TB and FB produce a blue fluorescence of the cytoplasm at this same wavelength. Therefore, NY may be combined with TB or FB in double-labeling experiments demonstrating the existence of axon collaterals. However, retrograde neuronal labeling with TB or FB requires a relatively long survival time, while NY requires a short survival time since NY migrates rapidly out of the retrogradely labeled neurons. This complicates double-labeling experiments since TB and FB must be injected first and NY later, a short time before the animal is sacrificed. We report a new yellow fluorescent tracer which labels mainly the nucleus and migrates much more slowly out of the retrogradely labeled neurons than NY. This new tracer can be used instead of NY in combination with TB or FB in double-labeling experiments and unlike NY can be injected at the same time as TB or FB. The new tracer is a diamidino compound (no. 28826) which is commerciallyFootnote 1 available. It will be referred to as Diamidino Yellow Dihydrochloride (DY·2HCl). According to the present study DY·2HCl is transported over long distances in rat and cat, and produces a yellow fluorescence of the neuronal nucleus at 360 nm excitation wavelength, resembling that obtained with NY. When combined with TB or FB, DY·2HCl is as effective as NY in double labeling of neurons by way of divergent axon collaterals.

This is a preview of subscription content, access via your institution.

Notes

  1. Distributed by Dr. Illing KG, Warthweg 14-18, Postfach 1150, D-6114 Groß-Umstadt, FRG

References

  • Bentivoglio M, Kuypers HGJM, Catsman-Berrevoets CE, Dann O (1979a) Fluorescent retrograde neuronal labeling in rat by means of substances binding specifically to adenine-thymine rich DNA. Neurosci Lett 12: 235–240

    Google Scholar 

  • Bentivoglio M, Van der Kooy D, Kuypers HGJM (1979b) The organization of the efferent projections of the substantia nigra in the rat. A retrograde fluorescent double labeling study. Brain Res 174: 1–17

    Google Scholar 

  • Bentivoglio M, Kuypers HGJM, Catsman-Berrevoets CE (1980a) Retrograde neuronal labeling by means of bisbenzimide and Nuclear Yellow (Hoechst S 769121). Measures to prevent diffusion of the tracers out of the retrogradely labeled neurons. Neurosci Lett 18: 19–24

    Google Scholar 

  • Bentivoglio M, Kuypers HGJM, Catsman-Berrevoets CE, Loewe H, Dann O (1980b) Two new fluorescent retrograde neuronal tracers which are transported over long distances. Neurosci Lett 18: 25–30

    Google Scholar 

  • Bharos TB, Kuypers HGJM, Lemon RN, Muir RB (1981) Divergent collaterals from deep cerebellar neurons to thalamus and tectum, and to medulla oblongata and spinal cord: Retrograde fluorescent and electrophysiological studies. Exp Brain Res 42: 399–410

    Google Scholar 

  • Björklund A, Skagerberg G (1979a) Simultaneous use of retrograde fluorescent tracers and fluorescence histochemistry for convenient and precise mapping of monoaminergic projections and collateral arrangements in the CNS. J Neurosci Meth 1: 261–277

    Google Scholar 

  • Björklund A, Skagerberg G (1979b) Evidence for a major spinal cord projection from the diencephalic A II dopamine cell group in the rat using transmitter-specific fluorescent retrograde tracing. Brain Res 177: 170–176

    Google Scholar 

  • Catsman-Berrevoets CE, Lemon RN, Verburgh CA, Bentivoglio M, Kuypers HGJM (1980) Absence of callosal collaterals derived from rat corticospinal neurons. Exp Brain Res 39: 443–440

    Google Scholar 

  • Catsman-Berrevoets CE, Kuypers HGJM (1981) A search for corticospinal collaterals to thalamus and mesencephalon by means of multiple retrograde fluorescent tracers in cat and rat. Brain Res 218: 15–33

    Google Scholar 

  • Flumerfelt BA, Gwyn DG (1974) The red nucleus of the rat: Its organization and interconnections. J Anat 118: 374–376

    Google Scholar 

  • Gwyn DG (1971) Acetylcholinesterase activity in the red nucleus of the rat. Effects of rubrospinal tractomy. Brain Res 35: 447–461

    Google Scholar 

  • Hayes NL, Rustioni A (1981) Descending projections from brainstem and sensorimotor cortex to spinal enlargements in the cat. Single and double retrograde tracer studies. Exp Brain Res 41: 89–107

    Google Scholar 

  • Huisman AM, Kuypers HGJM, Verburgh CA (1981) Quantitative differences in collateralization of the descending spinal pathways from red nucleus and other brain stem cell groups in rat as demonstrated with the multiple fluorescent retrograde tracer technique. Brain Res 209: 271–286

    Google Scholar 

  • Huisman AM, Kuypers HGJM, Verburgh CA (1982) Differences in collateralization of the descending spinal pathways from red nucleus and other brain stem cell groups in cat and monkey. In: Kuypers HGJM, Martin GF (ed) Descending pathways to the spinal cord. Progress in Brain Research, vol 57. Eisevier, Amsterdam New York, pp 185–217

    Google Scholar 

  • Huisman AM, Kuypers HGJM, Condé F, Keizer K (1983) Collaterals of rubrospinal neurons to the cerebellum in rat. A retrograde fluorescent double labeling study. Brain Res 264: 181–196

    Google Scholar 

  • Illert M, Fritz N, Aschoff A, Holländer H (1982) Fluorescent compounds as retrograde tracers compared with horseradish peroxidase (HRP). II. A parametric study in the peripheral motor system of the cat. J Neurosci Meth 6: 199–218

    Google Scholar 

  • Jacobson S, Trojanowsky JQ (1974) The cells of origin of the corpus callosum in rat, cat and rhesus monkey. Brain Res 74: 149–155

    Google Scholar 

  • Keizer K, Kuypers HGJM (1982) Distribution of corticospinal neurons in the cats sensorimotor cortex which neurons send collaterals to the tegmentum of the medulla oblongata and the lower pons. Neurosci Lett [Suppl] 10: S263

    Google Scholar 

  • Kuypers HGJM, Maisky VA (1975) Retrograde axonal transport of horseradish peroxidase from spinal cord to brain stem cell groups in the rat. Neurosci Lett 1: 9–14

    Google Scholar 

  • Kuypers HGJM, Bentivoglio M, Catsman-Berrevoets CE, Bharos TB (1980) Double retrograde neuronal labeling through divergent axon collaterals, using two fluorescent tracers with the same excitation wavelength which label different features of the cell. Exp Brain Res 40: 383–392

    Google Scholar 

  • Kuzuhara S, Kanazawa I, Nakanishi T (1980) Topographical localization of the Onuf's nuclear neurons innervating the rectal and vesical striated sphincter muscles: A retrograde fluorescent double labeling in cat and dog. Neurosci Lett 16: 125–130

    Article  CAS  PubMed  Google Scholar 

  • Martin GF, Cabana T, Humbertson AO (1981) Evidence for a lack of distinct rubrospinal somatotopy in the North American opossum and for collateral innervation of the cervical and lumbar enlargements by single rubral neurons. J Comp Neurol 201: 255–263

    Google Scholar 

  • Murray HM, Gurule ME (1979) Origin of the rubrospinal tract of the rat. Neurosci Lett 14: 19–23

    Google Scholar 

  • Niijima K, Yoshida M (1982) Electrophysiological evidence for branching nigral projections to pontine reticular formation, superior colliculus and thalamus. Brain Res 239: 279–282

    Google Scholar 

  • Wise SP, Jones EG (1976) The organization and postnatal development of the commissural projection of the rat somatic sensory cortex. J Comp Neurol 168: 313–344

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Supported in part by Grant 13-46-91 of FUNGO/ZWO Dutch Organization for Fundamental Research in Medicine

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Keizer, K., Kuypers, H.G.J.M., Huisman, A.M. et al. Diamidino yellow dihydrochloride (DY·2HCl); a new fluorescent retrograde neuronal tracer, which migrates only very slowly out of the cell. Exp Brain Res 51, 179–191 (1983). https://doi.org/10.1007/BF00237193

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00237193

Key words

  • Fluorescent retrograde tracers
  • Diamidino Yellow·2HCl
  • Retrograde double labeling