Highlights
- •Immunohistochemistry was combined with anterograde labelling to identify extrinsic nerve endings in the bladder.
- •This approach distinguished spinal afferent and autonomic efferent (motor) nerve endings in the bladder.
- •Morphology alone could not reliably distinguish simple and branching endings of afferent and autonomic neurons.
Abstract
The bladder is innervated by axons of sympathetic and parasympathetic efferent nerves,
and by spinal afferent neurons. The objective was to characterise anatomically and
immunohistochemically the terminal endings of sensory and autonomic motor nerve endings
in wholemount preparations of the mouse bladder. We used both anterograde labelling
of pelvic and hypogastric nerves ex vivo and anterograde labelling from lumbosacral dorsal root ganglia (DRG) in vivo in male and female mice. These were combined with immunohistochemistry for major
markers of sensory, sympathetic and parasympathetic nerves. Selective labelling of
spinal afferent endings following dextran biotin-labelling from DRGs in vivo showed no co-localisation of VAChT or TH in sensory terminals in the detrusor and
suburothelial plexus. Biotinamide was applied ex vivo to nerve trunks arising in the pelvic ganglion and running towards the bladder. Among
the filled axons, 38% of detrusor fibres and 47% of suburothelial axons were immunoreactive
for calcitonin-gene related peptide (CGRP). Vesicular acetylcholine transporter (VAChT)
immunoreactivity was present in 26% of both detrusor and suburothelial axons. For
tyrosine hydroxylase (TH), the proportions were 15% and 17%, respectively. Three major
morphological types of CGRP-immunoreactive nerve endings were distinguished in the
bladder wall: simple, branching and complex. VAChT-immunoreactive parasympathetic
axons had simple and branching endings; TH immunoreactive axons all had simple morphologies.
Our findings revealed that different subtypes of sensory and autonomic nerve endings
can be reliably identified by combining anterograde labelling ex vivo with specific immunohistochemical markers, although morphologically some of these
types of endings were indistinguishable.
Keywords
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Article info
Publication history
Published online: June 30, 2020
Accepted:
June 27,
2020
Received in revised form:
June 26,
2020
Received:
May 5,
2020
Identification
Copyright
© 2020 Elsevier B.V. All rights reserved.