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Featured Article: Selective ablation of IB4+ primary afferent neurons reduces mechanical and cold hyperalgesia in an EAE mouse model of multiple sclerosis
Nguyen KL, Lamerand SR, Deshpande RP, Taylor BK (2021) Featured Article: Selective ablation of IB4+ primary afferent neurons reduces mechanical and cold hyperalgesia in an EAE mouse model of multiple sclerosis. Targeting Trends 22
Related Products: IB4-SAP (Cat. #IT-10), Blank-SAP (Cat. #IT-21)
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A spinal microglia population involved in remitting and relapsing neuropathic pain
Kohno K, Shirasaka R, Yoshihara K, Mikuriya S, Tanaka K, Takanami K, Inoue K, Sakamoto H, Ohkawa Y, Masuda T, Tsuda M (2022) A spinal microglia population involved in remitting and relapsing neuropathic pain. Science 376(6588):86-90. doi: 10.1126/science.abf6805
Objective: To investigate pain recovery mechanisms.
Summary: The authors reveal a mechanism for the remission and recurrence ofneuropathic pain, providing potential targets for therapeutic strategies.
Usage: The dose of CTB-SAP and IB4-SAP was 8 ug/10 uL, diluted in PBS.
Related Products: CTB-SAP (Cat. #IT-14), IB4-SAP (Cat. #IT-10)
Contribution of small diameter non-peptidergic primary afferent neurons to central neuropathic pain in a new, more clinically relevant mouse model of multiple sclerosis
Nguyen KL, Lamerand SR, Deshpande RP, Taylor BK (2021) Contribution of small diameter non-peptidergic primary afferent neurons to central neuropathic pain in a new, more clinically relevant mouse model of multiple sclerosis. Neuroscience 2021 Abstracts P377/07. Society for Neuroscience, Virtual.
Summary: Over 50% of multiple sclerosis (MS) patients suffer from neuropathic pain (MSNP). Current treatments give inadequate relief due to incomplete understanding of underlying mechanisms. Recent electrophysiological recordings of primary afferent neurons (PAN) in the dorsal root ganglion (DRG) following experimental autoimmune encephalomyelitis (EAE), a mouse model of MS, revealed increased afterhyperpolarization in small-diameter fibers. These data form the premise of our goal to understand the contribution of small-diameter (peptidergic or non-peptidergic) PANs to MSNP. Arguably the most common method to induce EAE is administration of myelin oligodendrocyte glycoprotein (MOG) to generate an autoimmune response targeting the myelin sheath. A MOG fragment is typically given with two adjuvants: complete Freund’s adjuvant (CFA) to boost immunogenicity and pertussis toxin (PTX) to breakdown the blood-brain barrier and facilitate CNS immune cell infiltration. However, PTX can disrupt G-protein coupled receptors, cause pain, and alter autoimmune response gene expression. In 10-week-old C57BL/6 mice, we conducted the first rigorous comparison of a classic PTX EAE model with the novel non-PTX (nPTX) EAE model. We found that both PTX and nPTX EAE mouse models showed the same degree of: 1) motor deficits; 2) plantar hindpaw mechanical and cold hypersensitivity (except cold hypersensitivity resolved more quickly after PTX EAE than nPTX EAE); and 3) lumbar spinal cord demyelination. Unlike most rodent models of MS including PTX EAE, the nPTX EAE group exhibited somatosensory cortex demyelination, a core feature of MS in human patients and cold hypersensitivity. We suggest nPTX EAE to be the most clinically relevant rodent model available to study not only MSNP, but MS in general. To evaluate the contribution of peptidergic and non-peptidergic neurons to MSNP, we induced nPTX EAE. After 12 days we administered capsaicin (10µg/mouse, i.t.) or IB4-saporin (1.5µg/mouse, i.t.) to primarily ablate peptidergic or nonpeptidergic C-fibers, respectively. Ablation efficacy was successfully confirmed with dramatic loss in DRG of TRPV1/CGRP immunoreactivity (peptidergic C-fibers) following capsaicin, and IB4 immunoreactivity (nonpeptidergic C-fibers) following IB4-saporin. IB4-saporin, but not capsaicin, partially reduced mechanical hypersensitivity and reversed cold hypersensitivity within 9 days. These data suggest nonpeptidergic but not peptidergic C-fibers contribute to MSNP. Our next studies will use genetic knockout, chemogenetic, and optogenetic strategies using MrgprdCreER mice to modulate the activity of nonpeptidergic C-fibers.
Related Products: IB4-SAP (Cat. #IT-10)
Nociception impedes grasping recovery in the spinal cord injured rat.
Walker J, Kim T, Giszter S, Detloff MR (2021) Nociception impedes grasping recovery in the spinal cord injured rat. Neuroscience 2021 Abstracts P372.04. Society for Neuroscience, Virtual.
Related Products: IB4-SAP (Cat. #IT-10)
TRP channels in cancer pain
de Almeida AS, Bernardes LB, Trevisan G (2021) TRP channels in cancer pain. Eur J Pharmacol 904:174185. doi: 10.1016/j.ejphar.2021.174185 PMID: 34015320
Objective: To describe the role of TRP vanilloid 1 (TRPV1) and TRP ankyrin 1 (TRPA1) involved in cancer pain mechanisms.
Summary: Several studies have revealed that the administration of TRPV1 or TRPA1 agonists/antagonists and TRPV1 or TRPA1 knockdown reduced sensitivity to nociception in cancer pain models. Thus, TRP channels are potential targets for managing cancer-related pain syndromes.
Usage: Ablation of IB4 (+) neurons.
Related Products: IB4-SAP (Cat. #IT-10)
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Studying human nociceptors: from fundamentals to clinic
Middleton SJ, Barry AM, Comini M, Li Y, Ray PR, Shiers S, Themistocleous AC, Uhelski ML, Yang X, Dougherty PM, Price TJ, Bennett DL (2021) Studying human nociceptors: from fundamentals to clinic. Brain 144(5):1312-1335. doi: 10.1093/brain/awab048
Summary: The authors injected 5 µg of IB4-SAP into the sciatic nerve in the left thigh. Lesioned animals displayed attenuated NGF-induced hyperalgesia, as well as differences in other pain-model markers.
Related Products: IB4-SAP (Cat. #IT-10)
Antiplexin D1 antibodies relate to small fiber neuropathy and induce neuropathic pain in animals
Fujii T, Lee EJ, Miyachi Y, Yamasaki R, Lim YM, Iinuma K, Sakoda A, Kim KK, Kira JI (2021) Antiplexin D1 antibodies relate to small fiber neuropathy and induce neuropathic pain in animals. Neurol Neuroimmunol Neuroinflamm 8(5):e1028. doi: 10.1212/NXI.0000000000001028
Summary: NeP patient-derived plexin D1-IgG selectively binds to isolectin B4-positive unmyelinated C-fiber type small DRG neurons that sense mechanical pain.
Related Products: IB4-SAP (Cat. #IT-10)
Development of a novel pipette tip-aided cell cloning method for the effective isolation of genome-edited porcine cell
Sato M, Saitoh I, Akasaka E, Inada E (2021) Development of a novel pipette tip-aided cell cloning method for the effective isolation of genome-edited porcine cell. OBM Genetics 5(1):16. doi: 10.21926/obm.genet.2101126
Summary: Isolation of clonal cells from a single colony is an essential step in the process of obtaining pure populations of stably-transfected clones after gene transfer and the subsequent drug selection. In the present study, a novel, simple, and non-invasive technique for the isolation of cells from single colonies using a disposable pipette tip was developed.
Usage: A toxin-based, drug-free selection system involving IB4-SAP was employed in the present study.
Related Products: IB4-SAP (Cat. #IT-10)
A high efficacy selection method for transfected cells utilizing recombinant isolectin B4-saporin
Galvan MA, Shramm PA, Bouajram R, Lappi DA, Ancheta LR (2019) A high efficacy selection method for transfected cells utilizing recombinant isolectin B4-saporin. Neuroscience 2019 Abstracts 794.10. Society for Neuroscience, Chicago, IL.
Summary: Transfection protocols often rely on the use of antibiotics for the selection of transfected cells and has become the accepted approach for in vitro research and therapeutic applications. Antibiotics have several shortcomings such as cost, continuous use, and harmful effects — even on the transfected cell population. In addition, selection pressures are often inefficient and fail to provide a population of cells that express the gene of interest (GOI) at high levels. We have used three separate GOI’s to select for solely high-expressing transfectants using targeted toxin selection pressure. Normal Rat Kidney Cells (KNRK) were individually transfected to express green fluorescent protein (GFP), melanopsin or the low-affinity nerve growth factor receptor (p75) using an innovative new transfection delivery vector called pGEI. The results from various assays were utilized to visually determine the expression rate and pattern of the targeted toxin selection method. Melanopsin and p75 — a photopigment and nerve growth factor, respectively — were of great interest to express in our transfected cells as a means to study their role in the development and function of neurons. The delivery vector, pGEI, removes resident Galalpha(1-3)Gal epitopes from non- human mammalian cell surfaces. This residue is the target of recombinant Isolectin B4 – Saporin (IB4-SAP), a selective targeted toxin. IB4-SAP is extremely potent, with an EC50 in the low picomolar range for alpha-D-galactopyranoside expressing cells in vitro. The cells with the highest expression of the inserted vector, and therefore the GOI, will have these residues removed. Those that fail to express the vector or do not express the vector in high enough amounts, will not have all the residues removed, and will be targeted and eliminated via IB4-SAP. This method of selection provides a means of purifying the highest- expressing transfected populations using a more cost-effective and time-saving approach.
Related Products: IB4-SAP (Cat. #IT-10)
Role of nociceptive afferent input on forelimb reaching and grasping behaviors in the spinal cord injured rat
Walker JR, Ong A, Detloff MR (2019) Role of nociceptive afferent input on forelimb reaching and grasping behaviors in the spinal cord injured rat. Neuroscience 2019 Abstracts 572.09. Society for Neuroscience, Chicago, IL.
Summary: Individuals with spinal cord injury (SCI) suffer a loss of motor and sensory function. The current standard of care to recover fine motor control is rehabilitation focused on a combination of range of motion, aerobic, and strength training (ST). However, limited research has been conducted to determine the role of nociceptive afferent inputs from muscle on spinal plasticity and/or recovery of function. Using a rodent model of SCI strength training rehabilitation, we determined that motor training not only improves forelimb strength and fine motor function but also can modulate the development of neuropathic pain, suggesting that improvements in reaching and grasping may be due, in part, to plasticity of nociceptive afferents. To further explore this, Sprague-Dawley rats received injections of rIB4-conjugated saporin, mu p75-conjugated saporin or unconjugated (vehicle) into the cervical dorsal root ganglia unilaterally to eliminate non-peptidergic and peptidergic nociceptors. There is an uninjured cohort and a group with unilateral C5 SCI. Von Frey and Hargreaves’ tests were performed at baseline and several time points post-injection to assess the effcacy of the nociceptive elimination. Several measures of forelimb strength were recorded over time including the isometric pull task, a single pellet retrieval task and the Montoya staircase test. To confirm the depletion of peptidergic and non-peptidergic nociceptors following saporin injection and/or SCI, cervical DRGs and spinal cords were stained with antibodies against CGRP and isolectin-B4. An understanding of the role of nociceptors in spinal plasticity and functional motor and sensory recovery of SCI patients will guide future research and refine rehabilitation strategies to further improve their quality of life.
Related Products: IB4-SAP (Cat. #IT-10), mu p75-SAP (Cat. #IT-16)