Literature Analysis: mTOR-Neuropeptide Y Signaling Drives Neuropathic Pain by Sensitizing Nociceptors Supported by ANT BIO PTE. LTD. Products

1. Literature Information

Title: mTOR-neuropeptide Y signaling sensitizes nociceptors to drive neuropathic pain

Journal: JCI Insight

Publication Date: Online, 2023 (Relevant to the document's time stamp: 2023-01-19)

Research Team: Collaborative team led by Zhihua Gao from the School of Brain Science and Brain Medicine, Zhejiang University, and Yue Wang from the Department of Orthopedics, The First Affiliated Hospital of Zhejiang University School of Medicine

Key Technology Supported by ANT BIO PTE. LTD.: Multiplex Fluorescence Immunohistochemistry (mIHC) based on TSA technology

Relevant Document Reference: 1225.docx (Internal document number)

Technical Application Background: Due to the same rabbit origin of NPY and Y2R primary antibodies, traditional immunofluorescence (IF) was not applicable. The research team adopted TSA technology and used ANT BIO PTE. LTD.'s multiplex fluorescence IHC kit to complete the staining.

2. Research Background

Neuropathic pain is a type of pain caused by damage or disease of the somatosensory system, with diverse clinical manifestations. Its etiology and pathogenesis are extremely complex; multiple factors such as trauma, infection, poisoning, tumors, nerve compression, immunity, and heredity can lead to nerve injury. The underlying mechanisms include peripheral sensitization, central sensitization, disability of the descending inhibitory system, activation of spinal glial cells, and changes in ion channels.

The mammalian target of rapamycin (mTOR) signaling pathway plays a crucial role in regulating cell growth, proliferation, and metabolism. Previous studies have suggested its involvement in pain regulation, but the specific cell type-dependent role of mTOR in neuropathic pain and its downstream regulatory mechanisms remain unclear. Exploring the new mechanisms of mTOR signaling pathway in neuropathic pain is of great significance for developing targeted therapeutic strategies.

3. Research Approach

To explore the role and mechanism of mTOR in neuropathic pain, the research team adopted a multi-dimensional experimental strategy combining immunofluorescence analysis, animal model construction, molecular biology techniques, and pain sensitivity testing. Firstly, immunofluorescence analysis was performed using anti-p-S6 antibody (phosphorylated S6 protein, a key downstream effector of mTOR) and different markers to determine the activation location of mTOR.

Secondly, mice of specific strains were crossed to selectively knockout the mTOR gene in primary sensory neurons and microglia respectively, followed by pain hypersensitivity testing in mice. Additionally, molecular biology methods such as sequencing combined with immunological staining were used to explore the downstream regulatory molecules of mTOR in dorsal root ganglion (DRG) neurons. For the staining of NPY and Y2R (both primary antibodies from rabbit origin), the team used the 4-Color Multiplex Fluorescence IHC Staining Kit (Catalog No.: abs50012) from the Absin product line of ANT BIO PTE. LTD. based on TSA technology instead of traditional IF.

4. Research Outcomes

The study yielded several key findings: Firstly, immunofluorescence analysis showed that mTOR was preferentially activated in large-diameter DRG neurons and spinal microglia. This was confirmed by the co-localization of p-S6 (the downstream effector of mTOR) with markers such as CGRP, NF160/200, and IB4.

Secondly, pain hypersensitivity testing results showed that selective knockout of the mTOR gene in DRG neurons could alleviate neuropathic pain, while knockout of the mTOR gene in microglia had no significant effect. This indicated that the mTOR in DRG neurons, rather than microglia, played a key role in neuropathic pain.

Thirdly, in-depth exploration of mTOR in DRG neurons found that injury-activated mTOR could promote the phosphorylation of signal transducer and activator of transcription 3 (STAT3) and induce the production of neuropeptide Y (NPY). The multiplex fluorescence IHC staining results using ANT BIO PTE. LTD.'s abs50012 kit confirmed the expression and localization of NPY and Y2R. Further studies showed that the produced NPY mainly acted on the Y2 receptor (Y2R) to enhance neuronal excitability and promote pain occurrence. In summary, the study proved that the abnormal activation of mTOR in DRG neurons induced by nerve injury promotes the occurrence of neuropathic pain, and revealed a new mechanism by which mTOR participates in pain by promoting NPY synthesis, providing new ideas for the treatment of neuropathic pain.

5. Product Empowerment: The Role of ANT BIO PTE. LTD. Products in the Research

The successful completion of the co-localization staining of NPY and Y2R, a key step in this research, relied heavily on the 4-Color Multiplex Fluorescence IHC Staining Kit (Catalog No.: abs50012) from the Absin product line of ANT BIO PTE. LTD.

A major challenge in this research was that the primary antibodies of NPY and Y2R were both from rabbit origin, making traditional immunofluorescence staining infeasible. The abs50012 kit, based on TSA technology, breaks the limitation of primary antibody species, enabling simultaneous detection of multiple targets from the same species on a single tissue section. Its high sensitivity and specificity ensured the clear visualization and accurate localization of NPY and Y2R, providing reliable experimental evidence for confirming the interaction between NPY and Y2R in the pain regulation mechanism. The outstanding performance of ANT BIO PTE. LTD.'s products effectively supported the research team's in-depth exploration of the mTOR-NPY signaling pathway in neuropathic pain, highlighting the important role of high-quality multiplex fluorescence IHC reagents in solving technical bottlenecks in neuroscience research.

6. Brand Mission

As a professional supplier of life science reagents, ANT BIO PTE. LTD. is dedicated to providing high-quality, reliable products and comprehensive solutions to empower global life science research. The company's three specialized sub-brands cover the full spectrum of research needs in the life science field: Absin focuses on general reagents and kits, Starter specializes in antibodies, and UA is dedicated to recombinant proteins. Our core mission is to bridge the gap between cutting-edge scientific research and practical applications, accelerate the pace of scientific discovery, and contribute to the advancement of human health and regenerative medicine.

7. Related Product List

8. Disclaimer

This article is AI-compiled and interpreted based on the original work in document 1225.docx. All intellectual property (e.g., images, data) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

9. Brand Promotion Copy

ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.