title: Spiny versus stubby: 3D reconstruction of human myenteric (type I) neurons.
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 mum(3), of spinies 2,316 +/- 545 mum(3)), 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.
authors.affiliation: Department of Anatomy I, University of Erlangen-Nuremberg, Krankenhausstrasse 9, 91054 Erlangen, Germany.
authors.forename: Tobias M
authors.initials: TM
authors.lastname: Lindig
authors.forename: Vinod
authors.initials: V
authors.lastname: Kumar
authors.forename: Ron
authors.initials: R
authors.lastname: Kikinis
authors.forename: Steve
authors.initials: S
authors.lastname: Pieper
authors.forename: Falk
authors.initials: F
authors.lastname: Schrödl
authors.forename: Winfried L
authors.initials: WL
authors.lastname: Neuhuber
authors.forename: Axel
authors.initials: A
authors.lastname: Brehmer
mesh_terms: D000328:Adult; D000368:Aged; D002454:Cell Differentiation; D004743:Enkephalin, Leucine / Q000378:metabolism; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D009197:Myenteric Plexus / Q000166:cytology / Q000648:ultrastructure*; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure*; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
chemical_list: D014660:Vasoactive Intestinal Peptide; D004743:Enkephalin, Leucine