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NeuN Expression in Spinal Neurons Projecting to the Cerebellum

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Abstract

We studied the distinctive features of NeuN immunolabeling in the neurons located in the four cat spinal cord structures sending projections to the cerebellum: Clarke’s nucleus and border cells (L4 segment), central cervical nucleus (C3 segment), and Stilling’s nucleus (S2 segment). Using morphometric and densitometric analysis, we demonstrated that all the neurons of interest share a striking feature, namely an extremely low level of cytoplasmic NeuN immunolabeling against a high level of nuclear immunolabeling of this neuronal nuclear antigen, commonly used as a neuronal marker. The neuronal soma size averaged 1000–1850 µm2, which is comparable to another type of large neurons detected in slices, motoneurons (1140–1660 µm2). Therefore, we used motoneuronal populations in the corresponding segments to compare the magnitude of their optical density. The relative optical density of the neurons of interest was several times lower than that of the motoneurons (0.060 ± 0.030 vs 0.330 ± 0.127). No significant differences in optical density were revealed between the above four spinal cord structures. Given that all these cell populations are morphologically unique and similar, we believe that the features of NeuN expression can be used as a simple tool to visualize neurons sending projections to the cerebellum. This can be instrumental both in targeted morphological examination and in histological control after experimental exposures.

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ACKNOWLEDGMENT

The authors are grateful to P.E. Musienko for the assistance in working with a part of animals.

Funding

This study was supported by the Russian Science Foundation (project No. 21-15-00235).

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Authors and Affiliations

  1. Pavlov Institute of Physiology, Russian Academy of Sciences, St. Petersburg, Russia

    A. A. Veshchitskii, N. V. Pavlova, P. Yu. Shkorbatova, N. I. Nikitina & N. S. Merkulyeva

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  1. A. A. Veshchitskii
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  2. N. V. Pavlova
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  3. P. Yu. Shkorbatova
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  4. N. I. Nikitina
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Contributions

Experimental design (N.S.M.), histological preparations (A.A.V., P.Yu.Sh., N.I.N., N.V.P.), data collection and processing (N.S.M., A.A.V., P.Yu.Sh., N.V.P.), writing and editing the manuscript (N.S.M., A.A.V., P.Yu.Sh., N.V.P.).

Corresponding author

Correspondence to N. S. Merkulyeva.

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ETHICS APPROVAL

The work was carried out in compliance with the requirements of the European Communities Council Directive 1986 on the protection of animals used for experimental and other scientific purposes, as stated in the Guide for the Care and Use of Laboratory Animals (2010). Experimental protocols were approved by the Ethics Committee of Pavlov Institute of Physiology of the Russian Academy of Sciences (Meeting minutes dated 09/11/2015).

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The authors declare that they have no conflict of interest.

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Translated by A. Polyanovsky

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Russian Text © The Author(s), 2023, published in Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, 2023, Vol. 59, No. 6, pp. 522–531https://doi.org/10.31857/S0044452923060098.

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Veshchitskii, A.A., Pavlova, N.V., Shkorbatova, P.Y. et al. NeuN Expression in Spinal Neurons Projecting to the Cerebellum. J Evol Biochem Phys 59, 1974–1985 (2023). https://doi.org/10.1134/S0022093023060078

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