Mammalia

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    Protocols in Current Issue
    Characterization of Immunological Niches within Peyer’s Patches by ex vivo Photoactivation and Flow Cytometry Analysis
    Authors:  Adi Biram and Ziv Shulman, date: 03/20/2020, view: 198, Q&A: 0
    T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and ...
    An ex vivo Approach to Assess Mitochondrial ROS by Flow Cytometry in AAV-tagged Astrocytes in Adult Mice
    Authors:  Carlos Vicente-Gutierrez and Juan P. Bolaños, date: 03/20/2020, view: 392, Q&A: 0
    Mitochondrial reactive oxygen species (mROS) are naturally produced signalling molecules extremely relevant for understanding both health- and disease-associated biological processes. The study of mROS in the brain is currently underway to decipher ...
    Analyzing the Quenchable Iron Pool in Murine Macrophages by Flow Cytometry
    Authors:  Michael Riedelberger and Karl Kuchler, date: 03/20/2020, view: 128, Q&A: 0
    Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on ...
    Method for Measuring Mucociliary Clearance and Cilia-generated Flow in Mice by ex vivo Imaging
    Authors:  Eric Song and Akiko Iwasaki, date: 03/20/2020, view: 180, Q&A: 0
    Ex vivo biophysical measurements provide valuable insights into understanding both physiological and pathogenic processes. One critical physiological mechanism that is regulated by these biophysical properties is cilia-generated flow that ...
    Single-cell qPCR Assay with Massively Parallel Microfluidic System
    Authors:  Marta Prieto-Vila, Takahiro Ochiya and Yusuke Yamamoto, date: 03/20/2020, view: 182, Q&A: 0
    The single-cell transcriptome is the set of messenger RNA molecules expressed in one cell. It is extremely variable and changes according to external, physical and biochemical conditions. Due to sensitivity shortages, most of genetic studies use ...
    In vitro Measurement of CMP-Sialic Acid Transporter Activity in Reconstituted Proteoliposomes
    Authors:  James Cahill, Shivani Ahuja and Matthew R. Whorton, date: 03/20/2020, view: 215, Q&A: 0
    Nucleotide-sugar transporters (NSTs) facilitate eukaryotic cellular glycosylation by transporting nucleotide-sugar conjugates into the Golgi lumen and endoplasmic reticulum for use by glycosyltransferases, while also transferring nucleotide ...
    Optic Nerve Crush in Mice to Study Retinal Ganglion Cell Survival and Regeneration
    In diseases such as glaucoma, the failure of retinal ganglion cell (RGC) neurons to survive or regenerate their optic nerve axons underlies partial and, in some cases, complete vision loss. Optic nerve crush (ONC) serves as a useful model not only ...
    Shared Pheromonal Communication of Specific Fear Between Adult Sprague Dawley Rats
    Authors:  Samantha J. Carew and Abhinaba Ghosh, date: 03/20/2020, view: 149, Q&A: 0
    Rats are highly social animals, and mainly communicate with one another in two ways: through ultrasonic vocalizations and pheromones. Most research on pheromones has been dedicated those regarding sexual behavior, but more recently pheromones which ...
    Testing Bone Formation Induction by Calvarial Injection Assay in vivo
    Authors:  Narelle E McGregor, Ingrid J Poulton, Emma C Walker and Natalie A Sims, date: 03/20/2020, view: 258, Q&A: 0
    Bone formation occurs during embryogenesis, skeletal growth and during the process of skeletal renewal throughout life. In the process of bone formation, osteoblasts lay down a collagen-containing matrix, termed osteoid, which is gradually hardened ...
    Characterization of Immunological Niches within Peyer’s Patches by ex vivo Photoactivation and Flow Cytometry Analysis
    Authors:  Adi Biram and Ziv Shulman, date: 03/20/2020, view: 198, Q&A: 0
    [Abstract] T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and involves physical contacts between T and B cells. During these interactions, T cells endow the B cells ...
    An ex vivo Approach to Assess Mitochondrial ROS by Flow Cytometry in AAV-tagged Astrocytes in Adult Mice
    Authors:  Carlos Vicente-Gutierrez and Juan P. Bolaños, date: 03/20/2020, view: 392, Q&A: 0
    [Abstract] Mitochondrial reactive oxygen species (mROS) are naturally produced signalling molecules extremely relevant for understanding both health- and disease-associated biological processes. The study of mROS in the brain is currently underway to decipher their physiopathological roles and contributions in neurological diseases. Recent advances in this ...
    Analyzing the Quenchable Iron Pool in Murine Macrophages by Flow Cytometry
    Authors:  Michael Riedelberger and Karl Kuchler, date: 03/20/2020, view: 128, Q&A: 0
    [Abstract] Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on cellular iron homeostasis in macrophages remains challenging due to the limited amount of available ...
    Method for Measuring Mucociliary Clearance and Cilia-generated Flow in Mice by ex vivo Imaging
    Authors:  Eric Song and Akiko Iwasaki, date: 03/20/2020, view: 180, Q&A: 0
    [Abstract] Ex vivo biophysical measurements provide valuable insights into understanding both physiological and pathogenic processes. One critical physiological mechanism that is regulated by these biophysical properties is cilia-generated flow that mediates mucociliary clearance, which is known to provide protection against foreign particles and ...
    Single-cell qPCR Assay with Massively Parallel Microfluidic System
    Authors:  Marta Prieto-Vila, Takahiro Ochiya and Yusuke Yamamoto, date: 03/20/2020, view: 182, Q&A: 0
    [Abstract] The single-cell transcriptome is the set of messenger RNA molecules expressed in one cell. It is extremely variable and changes according to external, physical and biochemical conditions. Due to sensitivity shortages, most of genetic studies use bulk samples, providing only the average gene expression. Single-cell technologies have provided a ...
    In vitro Measurement of CMP-Sialic Acid Transporter Activity in Reconstituted Proteoliposomes
    Authors:  James Cahill, Shivani Ahuja and Matthew R. Whorton, date: 03/20/2020, view: 215, Q&A: 0
    [Abstract] Nucleotide-sugar transporters (NSTs) facilitate eukaryotic cellular glycosylation by transporting nucleotide-sugar conjugates into the Golgi lumen and endoplasmic reticulum for use by glycosyltransferases, while also transferring nucleotide monophosphate byproducts to the cytoplasm. Mutations in this family of proteins can cause a number of ...
    Optic Nerve Crush in Mice to Study Retinal Ganglion Cell Survival and Regeneration
    [Abstract] In diseases such as glaucoma, the failure of retinal ganglion cell (RGC) neurons to survive or regenerate their optic nerve axons underlies partial and, in some cases, complete vision loss. Optic nerve crush (ONC) serves as a useful model not only of traumatic optic neuropathy but also of glaucomatous injury, as it similarly induces RGC cell death ...
    Shared Pheromonal Communication of Specific Fear Between Adult Sprague Dawley Rats
    Authors:  Samantha J. Carew and Abhinaba Ghosh, date: 03/20/2020, view: 149, Q&A: 0
    [Abstract] Rats are highly social animals, and mainly communicate with one another in two ways: through ultrasonic vocalizations and pheromones. Most research on pheromones has been dedicated those regarding sexual behavior, but more recently pheromones which signal danger to conspecifics have been identified in rodents. In fact, rats are capable of ...
    Testing Bone Formation Induction by Calvarial Injection Assay in vivo
    Authors:  Narelle E McGregor, Ingrid J Poulton, Emma C Walker and Natalie A Sims, date: 03/20/2020, view: 258, Q&A: 0
    [Abstract] Bone formation occurs during embryogenesis, skeletal growth and during the process of skeletal renewal throughout life. In the process of bone formation, osteoblasts lay down a collagen-containing matrix, termed osteoid, which is gradually hardened by incorporation of mineral crystals. Although osteoblasts can be induced to differentiate and to ...
    Delayed Alternation Task for the Study of Spatial Working and Long Term Memory in Rats
    Authors:  Megi Hoxha and Marta Sabariego, date: 03/05/2020, view: 399, Q&A: 0
    [Abstract] Memory systems can hold previously presented information for several seconds, bridging gaps between discontinuous events. It has been previously demonstrated that the hippocampus and the medial entorhinal cortex (mEC) are necessary for memory retention over delay intervals in alternation tasks. Here we describe the delayed alternation task, a ...



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