Islam S, Gleber-Netto FO, Mulcahy CF, Glaun MDE, Srivastava S, Hunt PJ, Williams MD, Barbon CE, Spiotto M, Zhao W, Adebayo A, Akhter S, Xie T, Debnath KC, Sathishkumar HN, Myers B, Lothumalla S, Yama I, Burks JK, Gomez J, Rao X, Wang J, Woodman K, Mansour J, Arenkiel B, Osman KL, Haxton C, Lever TE, Hutcheson KA, Amit M (2024) Neural landscape is associated with functional outcomes in irradiated patients with oropharyngeal squamous cell carcinoma. Sco Transl Med 16:eabq5585. doi: 10.1126/scitranslmed.abq5585
Objective: To understand the correlation between neuronal changes and patient-reported and functional outcomes in patients with oropharyngeal squamous cell carcinoma (OPSCC).
Summary: Tumor enrichment of adrenergic (TH+) and CGRP+ sensory–afferent nerves correlated with poorer swallowing outcomes. Functional electromyography recordings showed correlations between growing (GAP43+) and immature cholinergic (ChAT+DCX+) nerves and denervation patterns in survivors of OPSCC. A murine model of radiation-induced dysphagia further confirmed that immature cholinergic and CGRP+ nerves were correlated with impaired swallowing. The results suggest that CGRP+ and ChAT+ neuronal signaling play distinct roles in tumor- and radiation-induced dysphagia in OPSCC and offer a comprehensive dataset on the neural landscape of OPSCC.
Usage: 500 μg in 3 μl of alpha-CGRP-streptavidin-saporin (CGRP-SAP; #IT-94) and anti-ChAT-SAP (#IT-42) was stereotactically injected into the intraganglionic region over 3 min.
Barioni NO, Beduschi RS, da Silva AV, Martins MG, Almeida-Francia CCD, Rodrigues SA, López DE, Gómez-Nieto R, Horta-Júnior JAC (2024) The role of the ventral nucleus of the trapezoid body in the auditory prepulse inhibition of the acoustic startle reflex. Hearing Research doi: 10.1016/j.heares.2024.109070 PMID: 38972084
Objective: To study the acoustic startle response through elimination of the ventral nucleus of the trapezoid body neurons via Anti-ChAT-SAP injection.
Summary: The elimination of ventral nucleus of the trapezoid body (VNTB) is used while measuring the auditory prepulse inhibition and acoustic startle response with and without this group of neurons to study their role in rats. It was found The VNTB stands as the sole identified source of cholinergic inputs to Cochlear root neurons.
Usage: Lesions in the VNTB were performed via a bilateral microinjection of a neurotoxin selective for cholinergic neurons, the anti-ChAT-saporin (IT-42, 0.25 ug/μl, 400 nL)
Nishioka M, Hata T (2024) Cholinergic interneurons in the dorsal striatum play an important role in the acquisition of duration memory. Eur J Neurosci 59(11):3061-3073. doi: 10.1111/ejn.16337 PMID: 38576223
Objective: To investigate duration-memory formation in the dorsal striatum.
Summary: Rats were sufficiently trained using a peak-interval 20s procedure and then infused with anti-choline acetyltransferase–saporin into the dorsal striatum to cause selective ablation of cholinergic interneurons. Lesions of the cholinergic cells show delayed memory acquisition and suggest dorsal striatum neurons play a role in new duration memory.
Usage: Each group of rats received aCSF or anti-choline acetyltransferase (ChAT)–saporin (Anti-ChAT-SAP, IT-42) at 0.5 μg/μL at 0.5 μl/min for 2 mins.
Loften A (2024) Cholinergic modulation of dopamine-related effects of ethanol in the rat. Univ Gothenburg Thesis.
Objective: To explore the role of acetylcholine and cholinergic interneurons (CIN) in ethanol-induced dopamine (DA) release and in the reinforcing effects of ethanol.
Summary: The author’s thesis supports an important role of accumbal CIN in ethanol ́s DA releasing and reinforcing effects, opening up for new potential pharmacological targetsfor treatments of alcohol use disorder.
Usage: in vivo rat model with depletion of accumbal CIN was developed utilizing anti-choline acetyltransferase-saporin (IT-42)
Loftén A, Cadeddu D, Danielsson K, Stomberg R, Adermark L, Söderpalm B, Ericson M (2024) Reduced alcohol consumption following ablation of cholinergic interneurons in the nucleus accumbens of wistar rats. Addict Biol 30(2):e70022. doi: 10.1111/adb.70022 PMID: 39936333
Objective: The aim of this study was to explore the functional role of accumbal Cholinergic interneurons (CIN) in alcohol-related behavior.
Summary: Rats with depleted CIN consumed significantly less alcohol than sham-treated controls. No differences in sucrose preference, motor activity, water intake or weight gain were noted between treatment groups, suggesting that the ablation selectively affected alcohol-related behavior. In conclusion, this study further supports a role for accumbal CIN in regulating alcohol-consummatory behavior.
Usage: Ablation of CIN was induced by local administration of Anti-ChAT-SAP (Cat #IT-42) bilaterally into the nucleus accumbens of male Wistar rats. Dose: 0.5 and 0.75 μg/ul.
Loftén A, Adermark L, Ericson M, Söderpalm B (2023) Regulation of ethanol-mediated dopamine elevation by glycine receptors located on cholinergic interneurons in the nucleus accumbens. Addict Biol 28(12):e13349. doi: 10.1111/adb.13349 PMID: 38017639
Objective: The aim of this study was to explore the role of glycine receptors (GlyRs) on cholinergic interneurons (CIN) in sustaining extracellular dopamine levels and in ethanol-induced dopamine release.
Summary: Alcohol use disorder is one of the major psychiatric disorders worldwide. Ethanol reward is one of the many factors contributing to the disorder. The rewarding and reinforcing properties of ethanol have been linked to activation of the mesolimbic dopamine system, an effect that involves glycine receptors (GlyRs) in the nucleus accumbens. The study suggests that CIN are not important for GlyR-mediated regulation of basal dopamine output, but that CIN ablation blunts the ethanol-induced dopamine release by reducing the release of GlyR agonists.
Usage: CIN were ablated by Anti-ChAT-SAP administered locally in the nucleus accumbens of male Wistar rats. Rabbit-IgG-SAP was used as a control. Microinfusion was performed unilaterally into the nAc at a concentration of 0.5 ug/ul at 0.05 ul/min for 10 min for a total of 0.5 ul.
Myers B, Islam S, Gleber Netto FO, Debnath KC, Srivastava S, Xie T, Akhter S, Adebayo AA, Miller J, Lothumalia S, Sathiskumar HN, Amit M (2023) Targeting nociceptive and cholinergic nerves in irradiated oropharyngeal cancer model reveals novel mechanism for dysphagia. Cancer Neuroscience Symposium
Objective: Explore the hypothesis that modulation of cholinergic (CHAT+) and nociceptive (CGRP+) neurons correlate with improved dysphagia.
Summary: Oropharyngeal squamous cell carcinoma is one of the most common types of head and neck cancer. Treatment for OPSCC includes surgery, radiation therapy, chemotherapy, or a combination of therapies. Despite advances in treatment, dysphagia (difficulty swallowing) is still a major burden for patients with OPSCC. The study established a novel murine OPSCC model to explore the role of nerves in dysphagia with cholinergic (CHAT) and nociceptive (CGRP) neurons playing an important role in swallowing outcomes. Targeting CHAT and CGRP could be a novel strategy for OPSCC patients with dysphagia.
Usage: 500 ng of Anti-ChAT-SAP was injected into the trigeminal ganglion in mice.
Loftén A, Adermark L, Ericson M, Söderpalm B (2021) An acetylcholine-dopamine interaction in the nucleus accumbens and its involvement in ethanol’s dopamine-releasing effect. Addict Biol 26(3):e12959. doi: 10.1111/adb.12959
Summary: Basal extracellular levels of dopamine within the nucleus accumbens are not sustained by muscarinic acetylcholine, whereas accumbal Cholinergic interneurons-ACh are involved in mediating ethanol-induced dopamine release.
Usage: Anti-ChAT-SAP or Rabbit IgG-SAP were infused at a flow rate of 0.05 μl/min for 10 min giving a total volume of 0.5 μl.
Ashkenazi SL, Polis B, David O, Morris G (2021) Striatal cholinergic interneurons exert inhibition on competing default behaviours controlled by the nucleus accumbens and dorsolateral striatum. Eur J Neurosci 53(7):2078-2089. doi: 10.1111/ejn.14873
Objective: To determine whether cholinergic interneurons contribute to the competition between both ventral and dorsolateral control systems.
Summary: Findings indicate a central role of cholinergic interneurons in regulating motivational impact on striatally controlled behaviors.
Usage: Anti-ChAT-SAP was diluted to 0.5 μg/μl in phosphate buffer saline and 0.5 μl were injected in each injection site.
Crevier-Sorbo G, Rymar, VV,Crevier-Sorbo R, Sadikot AF (2020) Thalamostriatal degeneration contributes to dystonia and cholinergic interneuron dysfuntion in a mouse model of huntington’s disease. Acta Neruopatho Commun 8(1):14. doi: 10.1186/s40478-020-0878-0 PMID: 32033588
Objective: To ablate the neurons of the Thalamostrial system (TS) to elucidate their role in the motor symptoms of Huntington’s Disease.
Summary: Huntington’s disease is an autosomal disorder characterized by involuntary movement and striatal neuronal loss. Glutaminergic input from the TS is implicated in disease progression and motor deficits. Anti-ChAT-SAP is used to ablate neurons in the Thalamostrial system to understand the role these neurons played in Huntington’s.
Usage: Mice underwent unilateral, striatal injections with either Anti-ChAT-SAP (IT-42) or Rabbit IgG-SAP (IT-35). The total volume and concentration of either saporin construct was the same (0.7 μL of 0.6 μg/μL solution).
