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Spiny versus stubby: 3D reconstruction of human myenteric (type I) neurons

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Abstract

We have compared the three-dimensional (3D) morphology of stubby and spiny neurons derived from the human small intestine. After immunohistochemical triple staining for leu-enkephalin (ENK), vasoactive intestinal peptide (VIP) and neurofilament (NF), neurons were selected and scanned based on their immunoreactivity, whether ENK (stubby) or VIP (spiny). For the 3D reconstruction, we focused on confocal data pre-processing with intensity drop correction, non-blind deconvolution, an additional compression procedure in z-direction, and optimizing segmentation reliability. 3D Slicer software enabled a semi-automated segmentation based on an objective threshold (interrater and intrarater reliability, both 0.99). We found that most dendrites of stubby neurons emerged only from the somal circumference, whereas in spiny neurons, they also emerged from the luminal somal surface. In most neurons, the nucleus was positioned abluminally in its soma. The volumes of spiny neurons were significantly larger than those of stubby neurons (total mean of stubbies 806 ± 128 μm3, of spinies 2,316 ± 545 μm3), and spiny neurons had more dendrites (26.3 vs. 11.3). The ratios of somal versus dendritic volumes were 1:1.2 in spiny and 1:0.3 in stubby neurons. In conclusion, 3D reconstruction revealed new differences between stubby and spiny neurons and allowed estimations of volumetric data of these neuron populations.

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Acknowledgments

The excellent technical assistance of Karin Löschner, Stephanie Link, Andrea Hilpert, Hedwig Symowski and Inge Zimmermann (all Erlangen) is gratefully acknowledged. We thank the Department of Molecular Plant Physiology (Erlangen) for helping with the deconvolution. We thank Mark Anderson (Boston), Ian Gibbins (Adelaide) and Christian Lauterbach (Erlangen) for their advice and feedback and Katie Mastrogiacomo (Boston) for carefully reading the manuscript. The study was supported by Deutsche Forschungsgemeinschaft (BR 1815/3) and National Institutes of Health (P41 RR13218 and U54 EB005149).

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Authors and Affiliations

  1. Department of Anatomy I, University of Erlangen-Nuremberg, Krankenhausstraße 9, 91054, Erlangen, Germany

    Tobias M. Lindig, Falk Schrödl, Winfried L. Neuhuber & Axel Brehmer

  2. Section of Experimental Magnetic Resonance of the CNS, University of Tuebingen, 72076, Tuebingen, Germany

    Tobias M. Lindig & Vinod Kumar

  3. Surgical Planning Laboratory, Department of Radiology, Harvard Medical School, Brigham and Women’s Hospital, Boston, MA, 02115, USA

    Tobias M. Lindig, Ron Kikinis & Steve Pieper

  4. Isomics Inc., Cambridge, MA, 02115, USA

    Steve Pieper

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  1. Tobias M. Lindig
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  2. Vinod Kumar
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  7. Axel Brehmer
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Correspondence to Axel Brehmer.

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Lindig, T.M., Kumar, V., Kikinis, R. et al. Spiny versus stubby: 3D reconstruction of human myenteric (type I) neurons. Histochem Cell Biol 131, 1–12 (2009). https://doi.org/10.1007/s00418-008-0505-9

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