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Retrograde tracing and neuropeptide immunohistochemistry of sensory neurones projecting to the cartilaginous distal femoral epiphysis of young rats
Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Medicine and Care, Pharmacology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Cell biology. Linköping University, Faculty of Health Sciences.
2000 (English)In: Cell and Tissue Research, ISSN 0302-766X, E-ISSN 1432-0878, Vol. 299, no 2, 193-200 p.Article in journal (Refereed) Published
Abstract [en]

Although cartilage is considered to be devoid of innervation, axons occur in the perichondrium and during development in cartilage canals, thereby having a relatively close spatial relationship to chondroblasts and chondrocytes. The present study locates the source of the sensory innervation of the femoral cartilaginous epiphyses of young rats and investigates whether the neuropeptide calcitonin gene-related peptide (CGRP) can influence chondrocytes. Retrograde tracing from the distal femoral epiphysis of young rats with Fast Blue (FB) showed labelled neuronal profiles in the L2-L5 dorsal root ganglia. Sample countings indicated that 50% of the FB-labelled neuronal profiles were located at the L3 level and 25% at the L4 level. The labelled neurones had diameters of 15-40 µm, with a peak at 25-30 µm. Immunohistochemistry showed that about 50% of the FB-labelled profiles contained CGRP. Together with the finding that CGRP influences bone cells to generate the second messenger cAMP, this result suggested the hypothesis that chondrocytes might be similarly influenced by CGRP. However, stimulation of cartilage slices with CGRP in vitro followed by an assay of the cAMP content did not provide support for this hypothesis. We conclude that primary sensory neurones containing CGRP project to the perichondrium and to cartilage canals of growing cartilage, and that exogenous CGRP does not elevate the cAMP content of cartilage slices in vitro.

Place, publisher, year, edition, pages
2000. Vol. 299, no 2, 193-200 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-24999DOI: 10.1007/s004410050017Local ID: 9419OAI: oai:DiVA.org:liu-24999DiVA: diva2:245323
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
In thesis
1. Sensory nerve fibres, neuropeptides and cartilage: Experimental studies in the rat
Open this publication in new window or tab >>Sensory nerve fibres, neuropeptides and cartilage: Experimental studies in the rat
2001 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

During development, maintenance and repair after injury, reciprocal interactions occur between the peripheral nervous system and the target tissues. In the Papers presented in this thesis, different aspects of such netvetarget influences between peripheral nerve fibres and skeletal tissues dtuing development and repair have been investigated in the rat. Developing rat cartilaginous bone primordia have a richly innervated and vascularised perichondriwn. In addition, larger bones exhibit cartilage canals containing blood vessels and putative sensory nerve fibres. Tills evoked the question if there is a nervous regulation of skeletal development. Denervation of the hind paws of young rats resulted in a deficient length growth but had no influence on the progress of secondary ossification. Since growth is mainly due to events in cartilage, cartilage projecting sensory neurones were identified and examined. Sensory neurones projecting to the rat cartilaginous distal femoral epiphyses were located mainly in the dorsal root ganglia (DRG) L3 and L4 and exhibited small or medium-sized diameters. A large proportion of these neurones contained the neuropeptides CGRP and/or SP. However, application of CGRP to cartilage explants in vitro did not stimulate the chondrocytes in terms of an elevation of the level of cyclic AMP. Another possibility would be that the neuropeptides affect the developmental growth of bone and chondrocytes indirectly via effects on the blood vessels. Experiments .involving tracing as above and eo-culture of labelled DRG neurones and perichondrial cells in combination with immunohistochenllstty or electrophysiology showed that the traced cultured neurones contained CGRP and/or SPin in vivo-like proportions and that most of the cartilage-projecting neurones were proton sensitive, This prompted the suggestion that the nerve fibres in the perichondrium and in cartilage canals might release CGRP and SP in response to local tissue acidosis, thereby promoting tissue homeostasis by monitoring the balance between vascular supply and metabolic load and by influencing angiogenesis and blood flow. Subsequently, possible target influences on the local presence of perichondrial sensory nerve fibres were investigated. Application of inflammation related cytokines (IL-1ß, IL-6 and LIF) affected sensory neurones eo-cultured with perichondrium- or skin-derived fibroblast-like cells in terms of survival and neurite growth. These effects were strongly influenced by the origin of the target cells. Finally, experiments using the adult rat patella showed that osteochondral defects heal spontaneously but incompletely and that healing is not accompanied by an increase of local nerve fibres at the times examined. In conclusion, the present results indicate that cartilagerelated sensory nerve fibres influence skeletal growth, that a high proportion of these neurones contain CGRP and SP, that CGRP does not activate chondrocytes in cartilage slices, that many cartilage related sensory nerve fibres are proton-sensitive· and likely have a vasoregulatory role, that inflammatory mediators have distinct effects on sensory neurones eo-cultivated with perichondrial cells and that healing of an osteochondral defect in the rat patella does not involve a local increase of cartilage-related nerve fibres.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2001. 70 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 712
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25639 (URN)10014 (Local ID)91-7373-141-1 (ISBN)10014 (Archive number)10014 (OAI)
Public defence
2001-12-14, Berzeliussalen, Universitetssjukhuset, Linköping, 13:00 (Swedish)
Opponent
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2012-08-23Bibliographically approved

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Edoff, KarinGrenegård, MagnusHildebrand, Claes

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