Retrograde and transganglionic transport of horseradish peroxidase-conjugated cholera toxin B subunit, wheatgerm agglutinin and isolectin B4 from Griffonia simplicifolia I in primary afferent neurons innervating the rat urinary bladder
Section snippets
Surgical procedures and tissue preparation
Experiments were performed on 16 adult male Sprague–Dawley rats (ALAB, Stockholm, Sweden) weighing 230–450 g. All efforts were made to minimize animal suffering and reduce the number of animal used. The experiments were carried out in accordance with the policy of the Society for Neuroscience on the use of animals in neuroscience research, and approved by the local animal research ethical committee. The animals were anaesthetized by an i.p. injection of chloral hydrate (30 mg/100 g body weight). A
Transganglionic transport of lectin–horseradish peroxidase conjugates
In the L6 and S1 spinal cord segments, HRP histochemical and lectin-immunofluorescence histochemical labelling was observed in all cases following injections of the lectin–HRP conjugates into the bladder wall. No labelling was found in the T13 or L1–2 spinal cord segments. There were no differences in the extent of labelling found by the two histochemical methods, although the HRP-histochemistry seemed to be more sensitive, showing a higher density of labelling than the
Discussion
Results from the present study show that CTB–, WGA– and B4–HRP were transported retrogradely to the T13–L2 and L6–S1 DRGs, and transganglionically to the L6–S1 spinal cord following injections into the bladder wall. Bladder afferent neurons in the L6–S1 DRGs, which had taken up and transported these lectin conjugates, were predominantly RT97-negative (81%). In addition, a large number of neurons that transported CTB–HRP also contained the neuropeptides CGRP (71%) and SP (36%).
Conclusion
The results of this study show that CTB–HRP is an effective retrograde and transganglionic tracer for visceral afferents from the urinary bladder, superior to both WGA–HRP and B4–HRP. It is transported mostly by neurons with unmyelinated fibres, containing the neuropeptides SP and CGRP. This is contrary to the situation for somatic primary afferent neurons in which CTB–HRP is transported almost selectively by myelinated fibres practically lacking these peptides.
Acknowledgements
This work was supported by the Swedish Medical Research Council, proj. no. 553. Dr Brita Robertson is greatly acknowledged for valuable advice and helpful discussions.
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- 1
Anatomy Department, Queen Mary and Westfield College, Mile End Road, London E1 4NS, U.K.
- 2
Department of Anatomy, Kyungpook National University, School of Medicine, Taegu 700-422, Korea.