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Authors: Kun-Han Lu, Zhongming Liu, Jaiyue Cao
Group: SPARC
Summary: Briefly, a gadolinium‐based contrast agent was mixed with the animal’s meal in order for chyme to appear “bright” in MRI scans, thereby delineating the gastric and intestinal volume. A multi‐slice MRI sequence was used to scan the GI volume with high spatial resolution, and a similar sequence with a smaller spatial coverage was used to scan antral contractions with high temporal resolution. Measurements of gastric functions and physiology included the overall change in GI volume, gastric emptying, forestomach volume, corpus volume, antral volume, antral contraction frequency, antral peristaltic wave velocity, antral contraction amplitude, pyloric opening size, intestinal filling and, indirectly, absorption.

Proper citation: Kun-Han Lu, Zhongming Liu, Jaiyue Cao 2019. Contrast-enhanced magnetic resonance imaging of gastric emptying and motility in rats. protocols.io https://dx.doi.org/10.17504/protocols.io.wvxfe7n Copy   

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Authors: Brett Hanzlicek, Dennis Bourbeau, Margot Damaser
Group: SPARC
Summary: In this study, we will be using Yorkshire Pigs, 50-70kg. The purpose of this study is to develop a tool to measure bowel fullness and activity. This system will eventually be able to provide continuous, wireless, battery−powered data and ultimately be translated for use in research and clinicalapplications of bowel function. We will measure porcine bowel compliance, pressures, and dimensions in acute live animal studies.

Proper citation: Brett Hanzlicek, Dennis Bourbeau, Margot Damaser 2021. SPARC Pig2 acute wired ColoMOCA implantation. protocols.io https://dx.doi.org/10.17504/protocols.io.bfxgjpjw Copy   

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Authors: Janet Keast, Peregrine Osborne, Nicole Wiedmann
Group: SPARC
Summary: This protocol is used for immunohistochemical visualisation of immediate early gene expression (c-Fos or Egr-1) in cryosections of rat lumbosacral spinal cord. Free-floating sections are processed in a double labelling protocol to distinguish immediate early gene expression in different neurochemical classes of spinal cord neurons:ChAT [choline acetyltransferase]: preganglionic neuronsTH [tyrosine hydroxyls]: dopaminergic neuronsPax2: inhibitory interneurons

Proper citation: Janet Keast, Peregrine Osborne, Nicole Wiedmann 2020. Immunohistochemical labelling of spinal cord neurons involved in bladder activity. protocols.io https://dx.doi.org/10.17504/protocols.io.bakkicuw Copy   

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Authors: Terry Powley, Zhenjun Tan, Matthew Ward
Group: SPARC
Summary: This protocol describes a process for the measurement of electrical stimulation-induced effects on duodenal motility in young adult Sprague-Dawley rats. Signals recorded from strain gauges attached to the proximal duodenal surface were used to measure the effect of stimulation by patch electrodes implanted at multiple sites across the rat stomach in an acute anesthetized preparation. The effect of stimulation was quantified as the ratio of various motility assessments during and after stimulation vs. before stimulation, and the data was used to create a functional map of duodenal motor response to localized gastric stimulation.

Proper citation: Terry Powley, Zhenjun Tan, Matthew Ward 2020. Measurement of duodenal motility using implanted strain gauges. protocols.io https://dx.doi.org/10.17504/protocols.io.2irgcd6 Copy   

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Authors: Elisa Gonzalez-Rothi, Yasin Seven, Latoya Allen, Marissa Ciesla, Gordon Mitchell
Group: SPARC
Summary: This protocol describes the immunofluorescent labeling technique used to identify adenosine 2a receptor expression in CtB-labelled phrenic motor neurons and within a defined region of interest surrounding phrenic motor neurons.

Proper citation: Elisa Gonzalez-Rothi, Yasin Seven, Latoya Allen, Marissa Ciesla, Gordon Mitchell 2019. SPARC Adenosine 2A Receptor Immunohistochemistry Protocol in Rat Tissues Labeled with Cholera Toxin B-fragment. protocols.io https://dx.doi.org/10.17504/protocols.io.2kfgctn Copy   

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Authors: Brett Hanzlicek, Ben Abelson, Margot Damaser, Dennis Bourbeau
Group: SPARC
Summary: This protocol for cat surgery to expose the badder for inactive device insertion.

Proper citation: Brett Hanzlicek, Ben Abelson, Margot Damaser, Dennis Bourbeau 2021. SPARC Cat acute UroMOCA implantation surgery. protocols.io https://dx.doi.org/10.17504/protocols.io.bf2pjqdn Copy   

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Authors: Nicole A. Pelot, J. Ashley Ezzell, Kara A. Clissold, Warren M. Grill
Group: SPARC
Summary: Protocol for collection, histological processing, and imaging of human vagus nerves.

Proper citation: Nicole A. Pelot, J. Ashley Ezzell, Kara A. Clissold, Warren M. Grill 2020. SPARC_Duke_Grill_OT2-OD025340_HumanVagusNerve_Collection_Histology_Microscopy. protocols.io https://dx.doi.org/10.17504/protocols.io.xcyfixw Copy   

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Authors: Seoeun Lee, Lori Zeltser
Group: Optical Clearing of Tissue, SPARC
Summary: This protocol describes how to stain Brown Adipose Tissue using iDisco protocol and visualize the result via confocal microscope.

Proper citation: Seoeun Lee, Lori Zeltser 2019. iDisco immunolabeling in brown adipose tissue (BAT). protocols.io https://dx.doi.org/10.17504/protocols.io.wqmfdu6 Copy   

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Authors: Terry Powley, Jennifer Mcadams, Robert Phillips
Group: SPARC
Summary: This protocol describes the methods used to trace and enable morphometric quantification of vagal afferent neurites in the rat stomach. A mixture of dextran conjugates was injected into the nodose ganglia of young adult Sprague-Dawley rats and after a survival period of 14 days for optimal tracer transport, stomachs were removed and processed as whole mounts. ABC-DAB was used to create a permanent gold-brown stain of all labeled afferent neurites. Subgroups of samples were also counterstained with either the panneuronal chromogen cuprolinic blue or with nNOS antibodies and steel gray chromogen to label nitrergic cells.

Proper citation: Terry Powley, Jennifer Mcadams, Robert Phillips 2019. High resolution labeling of vagal afferent fibers using Dextran-Biotin with counterstaining. protocols.io https://dx.doi.org/10.17504/protocols.io.2ipgcdn Copy   

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Authors: Billie Hunne, Martin Stebbing, Rachel M McQuade, John B Furness
Group: SPARC
Summary: Enteroendocrine cells are important regulators of gastrointestinal, digestive and metabolic function. Here we describe protocols for quantifying the relationship between nerve fibers and enteroendocrine cells in the rat gastric mucosa using immunohistochemistry and high resolution microscopy techniques

Proper citation: Billie Hunne, Martin Stebbing, Rachel M McQuade, John B Furness 2019. Immunohistochemistry and high resolution microscopy of rat gastric nerve fibers and their relationship with enteroendocrine cells. protocols.io https://dx.doi.org/10.17504/protocols.io.xz8fp9w Copy   

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Authors: Jeffrey Ardell, Marmar Vaseghi, Olujimi Ajijola
Group: SPARC

Proper citation: Jeffrey Ardell, Marmar Vaseghi, Olujimi Ajijola 2019. Pig-Stellate Stimulation Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.2nhgdb6 Copy   

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Authors: Seoeun Lee, Lori Zeltser
Group: SPARC
Summary: This protocol describes how to retrogradely label the Brown Adipose Tissue (BAT) - projecting sympathetic neruons in stellate ganglia (SG) using Cholera toxin subunit B (CTB) conjugated with Alexa Flour 555.

Proper citation: Seoeun Lee, Lori Zeltser 2019. Retrograde labeling of brown adipose tissue (BAT)-projecting sympathetic neurons with cholera toxin B (CTB). protocols.io https://dx.doi.org/10.17504/protocols.io.wjrfcm6 Copy   

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Authors: Grant Mccallum, Nathan Kostick, Joseph Marmerstein, Yang Zheng, Dominique Durand
Group: SPARC
Summary: We propose to develop a neural interface based on carbon nanotube (CNT) yarns with a size similar to large axons (10 µm) that has a mechanical and molecular compatibility with the nerve. The mechanical properties of the interface will be optimized by matching flexural rigidity of the implant to that of the glossopharyngeal nerve (GLN). Since the wire is very flexible, a custom-inserting technique has been developed for this purpose and tested in rats.  Following insertion, the signal-to-noise ratio and impedance of the wires will be measured for 3 months. The biocompatibility of the implant will be assessed using histological techniques.

Proper citation: Grant Mccallum, Nathan Kostick, Joseph Marmerstein, Yang Zheng, Dominique Durand 2021. Glossopharyngeal Nerve Chronic Recording In Anesthetized Rat. protocols.io https://dx.doi.org/10.17504/protocols.io.wssfeee Copy   

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Authors: Janet Keast, Peregrine Osborne
Group: SPARC
Summary: This protocol is used for studies of neurons containing retrograde tracer dye in adult rat dorsal root ganglia (DRG) or pelvic ganglia (PG; synonym, major pelvic ganglia). Cryosections of ganglia are mounted directly on slides for immunohistochemical staining. Antibodies have been selected to distinguish different neurochemical classes of PG neurons or to specifically identify subclasses of sensory neurons that are myelinated (immunoreactive for NF200 [neurofilament, 200 kD]) or nociceptive (immunoreactive for TRPV1).

Proper citation: Janet Keast, Peregrine Osborne 2019. Immunohistochemical analysis of ganglion neurons innervating the lower urinary tract [keast-001-stage03]. protocols.io https://dx.doi.org/10.17504/protocols.io.w3efgje Copy   

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Authors: Lauren Segers, Kendall Morris, Donald Bolser
Group: SPARC

Proper citation: Lauren Segers, Kendall Morris, Donald Bolser 2020. Morris USF Lab protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bci8iuhw Copy   

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Authors: Martha Campbell Thompson, Malavika Nair, Aidan Sullivan
Group: SPARC

Proper citation: Martha Campbell Thompson, Malavika Nair, Aidan Sullivan 2020. Vesselucida 360 Protocol for Segmenting and Analyzing Human Islet Microvasculature. protocols.io https://dx.doi.org/10.17504/protocols.io.bjfzkjp6 Copy   

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Authors: Nicole A. Pelot, J. Ashley Ezzell, Gabriel B. Goldhagen, Jake E. Cariello, Kara A. Clissold, Warren M. Grill
Group: SPARC
Summary: The protocol describes immunohistochemistry with anti-claudin-1, imaging, image segmentation, and image analysis methods to quantify human vagus nerve morphology.

Proper citation: Nicole A. Pelot, J. Ashley Ezzell, Gabriel B. Goldhagen, Jake E. Cariello, Kara A. Clissold, Warren M. Grill 2020. SPARC_Duke_Grill_OT2-OD025340_HumanVagusNerve_Claudin1IHC_Morphology. protocols.io https://dx.doi.org/10.17504/protocols.io.6fzhbp6 Copy   

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Authors: Thomas Taylor-Clark, Seol-Hee Kim
Group: SPARC
Summary: Mice are euthanized, perfused with fixative and the vagal ganglia extracted. Vagal ganglia are then cyosectioned. Slices are stained for protein expression using immunohistochemistry. Expression of specific proteins and reporter proteins isarevisualized using microscopy.

Proper citation: Thomas Taylor-Clark, Seol-Hee Kim 2019. Dissection and immunohistochemistry of mouse vagal ganglia. protocols.io https://dx.doi.org/10.17504/protocols.io.baumieu6 Copy   

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Authors: Max Novelli
Group: SPARC
Summary: Protocol for dataset “RNEL Bladder January 2019”Lower urinary track nerve responses to high-density epidural spinal cord stimulation ( RNEL January 2019 )

Proper citation: Max Novelli 2021. SPARC RNEL Bladder January 2019 protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.xszfnf6 Copy   

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Authors: Billie Hunne, Martin Stebbing, Rachel M. McQuade, John B. Furness
Group: SPARC
Summary: Enteroendocrine cells are important regulators of gastrointestinal, digestive and metabolic function. Here we describe protocols for identifying, mapping and characterising these cells in the rat gastric mucosa using immunohistochemistry and high resolution microscopy techniques

Proper citation: Billie Hunne, Martin Stebbing, Rachel M. McQuade, John B. Furness 2019. Immunohistochemistry and high resolution microscopy of rat gastric enteroendocrine cells. protocols.io https://dx.doi.org/10.17504/protocols.io.xeyfjfw Copy   

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