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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References
- Bladder activation: afferent mechanisms.Urology. 2002; 59: 43-50
- Pharmacologic perspective on the physiology of the lower urinary tract.Urology. 2002; 60: 13-22
- Neuropeptides in lower urinary tract function.Handb. Exp. Pharmacol. 2011; : 395-423
- Dorsal root ganglion neurons innervating pelvic organs in the mouse express tyrosine hydroxylase.Neuroscience. 2012; 223: 77-91
- Neurochemical characterization of extrinsic nerves in myenteric ganglia of the guinea pig distal colon.J. Comp. Neurol. 2015; 523: 742-756
- Neural control of the lower urinary tract.Compr. Physiol. 2015; 5: 327-396
- Tyrosine hydroxylase and vesicular acetylcholine transporter are coexpressed in a high proportion of intramural neurons of the human neonatal and child urinary bladder.Neurosci. Lett. 1999; 277: 157-160
- The distribution of vesicular acetylcholine transporter in the human male genitourinary organs and its co-localization with neuropeptide Y and nitric oxide synthase.Neurourol. Urodyn. 2000; 19: 185-194
- Innervation of the epithelium and lamina propria of the urethra of the female rat.Anat. Rec. 2019; 302: 201-214
- Characterization of axons expressing the artemin receptor in the female rat urinary bladder: a comparison with other major neuronal populations.J. Comp. Neurol. 2014; 522: 3900-3927
- Distribution of afferent axons in the bladder of rats.J. Neurocytol. 1998; 27: 141-155
- Sensory nerves in the mammalian urinary tract. An evaluation using light and electron microscopy.J. Anat. 1974; 117: 133-144
- Receptive properties of myelinated primary afferents innervating the inflamed urinary bladder of the cat.J. Neurophysiol. 1993; 69: 395-405
- Expression of the TRPM8-immunoreactivity in dorsal root ganglion neurons innervating the rat urinary bladder.Neurosci. Res. 2009; 65: 245-251
- Renaming all spinal autonomic outflows as sympathetic is a mistake.Auton. Neurosci. 2017; 206: 60-62
- Segmental distribution and peptide content of primary afferent neurons innervating the urogenital organs and colon of male rats.J. Comp. Neurol. 1992; 319: 615-623
- Primary afferent neurons containing calcitonin gene-related peptide but not substance P in forepaw skin, dorsal root ganglia, and spinal cord of mice.J. Comp. Neurol. 2015; 523: 2555-2569
- Location of interstitial cells and neurotransmitters in the mouse bladder.BJU Int. 2006; 97: 1332-1337
- Primary sensory neurones: neurofilament, neuropeptides, and conduction velocity.Brain Res. Bull. 1993; 30: 239-243
- The role of peptides in the regulation of the micturition reflex: an update.Gen. Pharmacol. 1991; 22: 1-24
- Tyrosine hydroxylase. The intitial step in noradrenaline biosynthesis.J. Biol. Chem. 1964; 239: 2910-2917
- Muscarinic acetylcholine receptor subtypes expressed by mouse bladder afferent neurons.Neuroscience. 2010; 168: 842-850
- Neurochemical characterization of extrinsic innervation of the guinea pig rectum.J. Comp. Neurol. 2004; 470: 357-371
- Vasoactive intestinal polypeptide and other preprovasoactive intestinal polypeptide-derived peptides in the female and male genital tract: localization, biosynthesis, and functional and clinical significance.Am. J. Obstet. Gynecol. 1995; 172: 1615-1631
- Cholinergic regulation of proliferation of the urothelium in response to E. coli lipopolysaccharide exposition.Int. Immunopharmacol. 2018; 56: 222-229
- Distribution and sub-types of afferent fibre in the mouse urinary bladder.J. Chem. Neuroanat. 2017; 79: 1-11
- Neurochemical features of boar lumbosacral dorsal root ganglion neurons and characterization of sensory neurons innervating the urinary bladder trigone.J. Comp. Neurol. 2013; 521: 342-366
- Choline acetyltransferase-like immunoreactivity in small diameter neurons of the rat dorsal ganglion.Neurosci. Lett. 1995; 198: 17-20
- Muscarinic agonists and antagonists: effects on the urinary bladder.Handb. Exp. Pharmacol. 2012; : 375-400
- Sensory fibers of the pelvic nerve innervating the rat's urinary bladder.J. Neurophysiol. 2000; 84: 1924-1933
- Identification of medium/high-threshold extrinsic mechanosensitive afferent nerves to the gastrointestinal tract.Gastroenterology. 2009; 137: 274-284
- Identifying unique subtypes of spinal afferent nerve endings within the urinary bladder of mice.J. Comp. Neurol. 2018; 526: 707-720
- Calcitonin gene-related peptide immunoreactivity in afferent neurons supplying the urinary tract: combined retrograde tracing and immunohistochemistry.Neuroscience. 1986; 18: 727-747
- Rapid anterograde and retrograde tracing from mesenteric nerve trunks to the guinea-pig small intestine in vitro.Cell Tissue Res. 1999; 295: 437-452
- Distribution of sacral afferent axons in cat urinary bladder.Am. J. Anat. 1973; 136: 305-313
- Unbiased classification of sensory neuron types by large-scale single-cell RNA sequencing.Nat. Neurosci. 2015; 18: 145-153
- Afferent and sympathetic innervation of the dome and the base of the urinary bladder of the female rat.Brain Res. Bull. 1992; 29: 651-658
- Characterization of mouse lumbar splanchnic and pelvic nerve urinary bladder mechanosensory afferents.J. Neurophysiol. 2008; 99: 244-253
- Effects of spinal cord injury on neurofilament immunoreactivity and capsaicin sensitivity in rat dorsal root ganglion neurons innervating the urinary bladder.Neuroscience. 1998; 83: 633-643
- Transduction sites of vagal mechanoreceptors in the guinea pig esophagus.J. Neurosci. 2000; 20: 6249-6255
- Intraganglionic laminar endings are mechano-transduction sites of vagal tension receptors in the guinea-pig stomach.J. Physiol. 2001; 534: 255-268
- Properties of the major classes of mechanoreceptors in the guinea pig bladder.J. Physiol. 2007; 585: 147-163
- Mechanotransduction and chemosensitivity of two major classes of bladder afferents with endings in the vicinity to the urothelium.J. Physiol. 2009; 587: 3523-3538
- Structure-function relationship of sensory endings in the gut and bladder.Auton. Neurosci. 2010; 153: 3-11
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.
