Quantitation of lower urinary tract afferents in 3D reconstruction of lumbosacral spinal cord sections v1

2021 ◽  
Author(s):  
John-Paul Fuller-Jackson ◽  
Peregrine B Osborne ◽  
Janet R Keast
Keyword(s):  
Spinal Cord ◽  
Urinary Tract ◽  
3D Reconstruction ◽  
Lower Urinary Tract ◽  
Single Image ◽  
Toxin B ◽  
Image Stack ◽  
Lumbosacral Spinal Cord ◽  
Cholera Toxin B ◽  
Lower Urinary

This protocol details the 3D reconstruction of the lumbosacral spinal cord using alternating cryosections, and then goes through the steps required to quantify lower urinary tract afferents. Using TissueMaker (MBF Bioscience), images of alternating sections can be ordered and aligned prior to the production of a single image stack. In Neurolucida 360 (MBF Bioscience), regions of interest can be defined within the image stack, and the bouton-like immunolabelling of cholera toxin B can be segmented. Once saved, this data can then be extracted using Neurolucida Explorer (MBF Bioscience).

Immunohistochemical labelling of lower urinary tract afferents in spinal cord v1

2021 ◽  
Author(s):  
Janet R Keast ◽  
Peregrine B Osborne ◽  
John-Paul Fuller-Jackson
Keyword(s):  
Spinal Cord ◽  
Urinary Tract ◽  
Cholera Toxin ◽  
Lower Urinary Tract ◽  
Double Labelling ◽  
Toxin B ◽  
Lumbosacral Spinal Cord ◽  
Cholera Toxin Subunit B ◽  
Subunit B ◽  
Lower Urinary

This protocol is used for immunohistochemical visualisation of cholera toxin subunit B within afferents innervating the lower urinary tract in cryosections of rat lumbosacral spinal cord. Free-floating sections are processed in a double labelling protocol to distinguish regions of innervation by these afferents. Cholera toxin B antibody [lower urinary tract afferents] Choline acetyltransferase antibody [preganglionic autonomic neurons and motoneurons]

Visualizing lower urinary tract afferent projections in the lumbosacral spinal cord in rats v1

2021 ◽  
Author(s):  
John-Paul Fuller-Jackson ◽  
Peregrine B Osborne ◽  
Janet R Keast
Keyword(s):  
Spinal Cord ◽  
Urinary Tract ◽  
3D Reconstruction ◽  
Lower Urinary Tract ◽  
Sprague Dawley ◽  
Lumbosacral Spinal Cord ◽  
Afferent Projections ◽  
Sprague Dawley Rats ◽  
Stage 1 ◽  
Lower Urinary

This collection includes the protocols required to map the lower urinary tract afferent projections to the lumbosacral spinal cord of male and female Sprague-Dawley rats. Afferents can be visualized using 3D reconstruction of alternating sections in TissueMaker (MBF Bioscience), or through the immunolabelling and clearing method, iDISCO. The following protocols are performed, regardless of endpoint: STAGE 1: Use of cholera toxin subunit B to label neural projections to lower urinary tract organs. STAGE 2: Intracardiac perfusion with fixative for anatomical studies. The next set of protocols pertain to the 3D reconstruction of spinal cord from alternating sections. STAGE 3: Immunohistochemical labelling of lower urinary tract afferents in spinal cord. STAGE 4: Quantitation of lower urinary tract afferents in 3D reconstruction of lumbosacral spinal cord sections For the visualization of lower urinary tract afferents in the intact spinal cord, skip Stages 3 and 4, and instead use Stage 5: STAGE 5: Immunolabelling and clearing of intact spinal cord for visualization of lower urinary tract afferents

Immunolabelling and clearing of intact spinal cord for visualization of lower urinary tract afferents v1

2021 ◽  
Author(s):  
Janet R Keast ◽  
Peregrine B Osborne ◽  
John-Paul Fuller-Jackson
Keyword(s):  
Spinal Cord ◽  
Urinary Tract ◽  
Lower Urinary Tract ◽  
Light Sheet ◽  
Lumbosacral Spinal Cord ◽  
Cholera Toxin Subunit B ◽  
Autonomic Neurons ◽  
Subunit B ◽  
Clearing Method ◽  
Lower Urinary

The whole-mount immunolabeling and clearing method (iDISCO) was used to visualize cholera toxin subunit B-labelled lower urinary tract afferents in the lumbosacral spinal cord of the rat. Imaging of spinal cord was performed on a light sheet microscope with a 12x lens. Concurrently, choline acetyltransferase identified preganglionic autonomic neurons and motoneurons within the spinal cord, which were used to confirm the rostrocaudal location of afferents.

scholarly journals Pathophysiology of the lower urinary tract and CNS

2013 ◽  
Vol 5 (5-S2) ◽  
pp. 126
Author(s):  
Christopher Chapple
Keyword(s):  
Spinal Cord ◽  
Central Nervous System ◽  
Nervous System ◽  
Urinary Tract ◽  
Lower Urinary Tract ◽  
Neural Control ◽  
Neural Mechanisms ◽  
The Other ◽  
Bladder Function ◽  
Lower Urinary

A number of aspects of neural control are potentially important inthe control of bladder function, including both sensory and motorand peripheral and central pathways. It is likely that a combinationof disorders of both central and peripheral neural mechanisms isimportant in the genesis of urgency and the other symptoms of theoveractive bladder (OAB). Given the number of potential pathwaysinvolved, potential pharmacologic targets for OAB exist in the CNS(central nervous system; cerebral cortex, midbrain, spinal cord)and periphery (LUT; lower urinary tract). Antimuscarinics are stillthe mainstay of OAB treatment, but there are also a number ofother potentially efficacious drugs that may also provide benefitagainst the neurologic components of OAB. This review discussesthe impact of neurological abnormalities on lower urinary tractsymptoms and the potential for treatments targeting these pathwaysto improve symptoms.

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