Nazmuddin M, Stammes MA, Klink PC, Vernes MK, Bakker J, Langermans JAM, van Laar T, Philippens IHCHM (2025) Stereotactic lesioning of cholinergic cells by injection of ME20.4 Saporin in the nucleus basalis of Meynert in a rhesus monkey (Macaca mulatta). J Neuropathol Exp Neurol nlaf081. doi: 10.1093/jnen/nlaf081 PMID: 40673943
Objective: To describe a procedure to inject ME20.4-SAP, an immunotoxin that specifically binds to and depletes cholinergic neurons stereotactically into the nucleus basalis of Meynert (NBM) of a rhesus monkey (Macaca mulatta).
Summary: A digital non-human primate brain atlas was co-registered to the brain of the monkey. A custom-designed cranial chamber was also implanted to the skull to guide the injection. The effects of the ME20.4-SAP injections were evaluated in vivo with PET-CT using [18F]-FEOBV as a radiotracer. This approach yielded reliable spatial accuracy and successful delivery of ME20.4-SAP into the NBM. This saporin-mediated selective destruction of cholinergic neurons in the NBM, using MRI-guidance and a cranial chamber, offers a promising method to study the pathophysiology of NBM degeneration and possible therapeutic interventions.
Usage: The first dose was chosen based on previous NBM lesioning works in common marmosets where infusing 1.4 μg ME20.4-SAP (Cat. #IT-15, in a concentration of 0.20 μg/μl) into each side of the NBM produced partial NBM depletion. At the second injection session, 5 μg ME20.4-SAP (in 0.5 μg/μl solution) was administered into each NBM side.
Ishtiaque S, Ahman S, Farooqui A, Siddiqui MH, Faridi SA (2025) Multifunctional nanoparticles to over come ABC transporter-mediated drug resistance in cancer: A short review. IJERT 13(6)
Objective: To examine innovative multifunctional nanoparticle strategies designed to overcome transporter-mediated efflux mechanism.
Summary: Authors analyze nanocarrier platforms enabling targeted co-delivery of chemotherapeutics with transporter inhibitors, gene-silencing approaches suppressing efflux pump expression, and stimuli-responsive systems exploiting tumor microenvironment features. The work highlights emerging technologies including nanobots and AI-designed nanoparticles while addressing translational challenges like protein corona formation and manufacturing scalability.
Usage: Lipid-SAP (EC16-1/saporin) was mentioned as one of the selected preclinical studies of nanoparticle platforms targeting ABC transporter-mediated drug resistance.
Noble EE, Harris RBS (2025) Leptin in the VMH contributes to the initial overconsumption of palatable diets by rats. Am J Physiol Endocrinol Metab 329(1):E1-E17. doi: 10.1152/ajpendo.00090.2025 PMID: 40418155
Objective: To determine whether leptin receptor–expressing neurons in the ventromedial hypothalamus (VMH) contribute to the initial overconsumption of a high-fat diet in rats.
Summary: Ablation of VMH leptin receptor–expressing neurons using Leptin-SAP prevented the early hyperphagic response to a high-fat diet in male rats but had no long-term impact on energy intake, body weight, or glucose clearance. These findings suggest VMH leptin signaling plays a key role in initiating, but not maintaining, diet-induced hyperphagia.
Usage: Leptin-SAP (IT-47) or Blank-SAP (IT-21) was stereotaxically injected into the VMH of male and female rats (20 ng in 80 nL) to ablate leptin receptor–expressing neurons. This targeted lesion confirmed the role of VMH leptin signaling in mediating early-phase overeating in response to a high-fat diet.
Bai J, Cheng K, Zhang N, Chen Y, Ni J, Wang Z (2025) Research advances in dysphagia animal models. Animal Model Exp Med doi: 10.1002/ame2.70054 PMID: 40566744
Objective: To summarize the establishment and evaluation of dysphagia animal models in stroke, Parkinson’s disease, and ALS, in three kinds of experimental animals, providing a basis for the selection of appropriate animal models of dysphagia.
Summary: There are very few studies of diseases such as stroke, PD, and ALS using other mammal and NHP dysphagia models. Moreover, there are substantial labor, cost, time, and ethics-related issues that limit the widespread use of these animal models in research. Nevertheless, large animal models serve as a crucial intermediary between rodent studies and clinical trials, significantly enhancing the translational potential of preclinical research findings.
Usage: Lori et al. used an ALS model created by intralingual injection of cholera toxin B conjugated to saporin (CTB-SAP) to induce apoptosis of sublingual motoneurons to study the effect of hypoglossal motor neuron death without many complications.
Szysiak N, Kosior-Korzecka U, longo V, Patkowski K, Greguła-Kania M, Nowakiewicz A, Bochniarz M,Junkuszew A (2025) Influence of neurokinin b, dynorphin a and kisspeptin-10 on in vitro gonadotropin secretion by anterior pituitary cells isolated from pubescent ewes. J Vet Res doi: 10.2478/jvetres-2025-0003
Objective: The aim of the study was to analyze the direct effect of the hypothalamic neuropeptides kisspeptin-10, neurokinin B, and dynorphin A on gonadotropin secretion by pituitary cells isolated from pubescent ewes.
Summary: Puberty is a multifactorial and complex process in animal development and in the case of livestock, timely attainment of sexual maturity contributes to increased reproductive efficiency, which leads to higher profitability. Studies revealed that kisspeptin, neurokinin B and dynorphin neuropeptides, collectively referred to as KNDy neuropeptides, are recognized as the key neuropeptides produced and secreted by the arcuate nucleus of the hypothalamus (ARC), and involved in the endocrine regulation of the onset of puberty. They all play roles in the endocrine regulation of the hypothalamic-pituitary-ovarian (HPO) axis in puberty. Kisspeptin-10, NKB and Dyn A had a direct impact on gonadotropin secretion by ovine pituitary cells. However, a detailed explanation of their role in gonadotropin secretion by the anterior pituitary gland in sheep and their impact on the regulation of the HPO axis during sexual maturation or in the pathomechanism of delayed puberty requires further studies.
Usage: Prepubertal ewes received 1 μL (0.7 μg) of NKB-SAP (NK3-SAP) [IT-63] or Blank-SAP (IT-21) injections aimed at the arcuate (ARC) nucleus to ablate neurons expressing NK3R.
Orciani C, Foret MK, Cuello AC, Carmo SD (2025) Long-term nucleus basalis cholinergic lesions alter the structure of cortical vasculature, astrocytic density and microglial activity in Wistar rats. Neurobiology of Aging 150:132-145. doi: 10.1016/j.neurobiolaging.2025.03.006 PMID: 40121723
Objective: To investigate the effects of the Basal forebrain cholinergic neurons (BFCNs) input on neurovascular unit (NVU) components.
Summary: To address this issue, the authors immunolesioned the nucleus basalis by administering injections of the cholinergic immunotoxin 192-IgG-SAP. Authors observed a significant reduction in cortical vesicular acetylcholine transporter-immunoreactive synapses. This was accompanied by changes in the diameter of cortical capillaries and precapillary arterioles, as well as lower levels of vascular endothelial growth factor A (VEGF-A). Additionally, the cholinergic immunolesion increased the density of cortical astrocytes and microglia in the cortex. The loss of nucleus basalis cholinergic input negatively impacts cortical blood vessels, NVU components, and microglia phenotype.
Usage: 192-IgG-SAP (2.6 mg/ml, IT-01) was injected at 0.5 μg/μl (1.0 μl/ hemisphere).
Kim P, Kumar V, Garner N, Jayasingh O, Roman G, Walters S, Vo T, Nguyen Q, Bowles J, Woodruff T, Inder W, Hunt J, Heyde I, Oster H, Rawashdeh O (2025) A systemic clock brake: Period1 stabilizes the circadian network under environmental stress. bioRxiv 2025.06.12.659230. doi: 10.1101/2025.06.12.659230
Objective: To investigate the role of the core circadian clock gene Period1 (Per1) in regulating light-induced circadian realignment and systemic physiological stability across central and peripheral tissues.
Summary: Per1-deficient mice showed accelerated behavioral, hormonal, and metabolic re-entrainment to shifted light-dark cycles, highlighting Per1’s role as a buffer that stabilizes circadian responses. Despite faster adaptation, Per1 deletion compromised SCN network coherence and increased peripheral metabolic phase instability.
Usage: Melanopsin (OPN4) was detected using Anti-Melanopsin (AB-N38) at a 1:2000 dilution to quantify ipRGCs in the retina and confirm that Per1-deficiency did not affect melanopsin-positive cell abundance.
Lauer LT, Decarie-Spain L, Hayes AMR, Suarez AN, Bashaw A, Klug ME, Kao AE, Cheng R, Rea JJ, Subramanian KS, Nourbash A, Donohue KN, Schier LA, Myers K, Kanoski SE (2025) The vagus nerve promotes memory via septo-hippocampal acetylcholine: Implications for obesity-induced cognitive dysfunction. bioRxiv 2025.06.11.659206.
Objective: To demonstrate that nutrient consumption promotes hippocampal-dependent memory function via vagus nerve-mediated acetylcholine (ACh) release in the dorsal hippocampus (HPCd) from medial septum (MS) neurons.
Summary: Results identify a neurobiological mechanism whereby nutrient consumption promotes memory function, and reveals that disruption of this vagal-brain signaling system mediates Western Diet-associated memory impairments.
Usage: The antineuronal immunotoxin 192-IgG-SAP, which has been validated as an agent that selectively ablates neurons that produce acetylcholine was injected in the medial septum; 200nL (0.16mg/mL) were infused in three different coordinates that span the medial septum.
Tolymbekova A, Lezina L (2025) CD320 receptor and vitamin B12 as potential targets for anti-cancer therapy. Int J Mol Sci 26(12):5652. doi: 10.3390/ijms26125652 PMID: 40565117
Objective: To develop therapies discriminating between healthy and cancerous cells to prevent unwanted toxicity of anticancer agents.
Summary: One possible method is to target proteins overexpressed in cancer but not in normal cells. CD320 is a receptor responsible for the uptake of the transcobalamin-bound fraction of vitamin B12 (cobalamin), which is necessary for DNA synthesis, and thus, cell proliferation. CD320 was shown to be overexpressed in many cancers and its potential role as an early cancer biomarker was confirmed in several studies.
Usage: In CD320-overexpressing HEK293 cells, a 2.5 nM concentration of monoclonal antibodies conjugated with saporin was enough to induce a 100% inhibition of proliferation, while in normal HEK293 cells, the same concentration of antibodies induced approximately 50% inhibition of cell proliferation.
Zhu M, Wu Y, Gao H, Qi F, Zhang X, Ran Y (2025) Differential regulation of mTOR activity in retinal ganglion cells underlies their distinct susceptibility to ischemia/reperfusion. Commun Biol 8(1):911. doi: 10.1038/s42003-025-08314-2 PMID: 40500296
Objective: To explore why intrinsically photosensitive retinal ganglion cells (ipRGCs) are more resistant to ischemia/reperfusion (I/R) injury than other RGC subtypes and to examine the role of mTOR signaling in this differential vulnerability.
Summary: ipRGCs exhibited higher mTOR activity and greater resistance to I/R injury compared to other RGCs. Rapamycin had cell-type–specific effects: it protected non-ipRGCs by increasing mTOR activity but suppressed mTOR in ipRGCs unless light was removed, revealing that light conditions critically influence mTOR-mediated neuroprotection.
Usage: Melanopsin (OPN4) was detected using Anti-Melanopsin (AB-N39) at a 1:2000 dilution to identify and quantify ipRGCs in retinal whole-mounts following ischemic injury.
