DOCSLIB.ORG

5986.Full.Pdf

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
5986.Full.Pdf The Journal of Neuroscience, October 1994, 14(10): 59885995 Behavioral, Biochemical, Histological, and Electrophysiological Effects of 192 IgG-Saporin Injections into the Basal Forebrain of Rats Gary L. Wenk,’ James D. Stoehr,’ George Quintana,’ Sherri Mobley,’ and Ronald G. Wiley2 ‘Arizona Research Laboratories, Division of Neural Systems, Memory and Aging, University of Arizona, Tucson, Arizona 85724 and 2DVAMC and Vanderbilt University, Nashville, Tennessee 37232 The behavioral, biochemical, histological, and electrophys- layed-alternation or passive avoidance tasks were not im- iological effects of a basal forebrain injection of saporin, a paired by the lesions. ribosome-inactivating protein, coupled to a monoclonal an- These data confirm the effectiveness and specificity of tibody against the low-affinity NGF receptor (192 IgG) were this novel lesioning tool and suggest that selective loss of investigated in adult rats. Within the basal forebrain region, NBM cholinergic cells is not sufficient to impair performance the low-affinity NGF receptor is exclusively expressed by in these behavioral tasks. cholinergic neurons in the medial septal area, diagonal band, [Key words: 192 IgG-saporin, nucleus basalis, EEG, basal and nucleus basalis magnocellularis (NBM). The presence forebrain, lesion, T-maze alternation, rats] of this receptor upon these cells confers a degree of spec- ificity to the 192 IgG-saporin that could not previously be Loss of neurons in the basal forebrain may play an important achieved by previous lesioning techniques, such as excit- role in the cognitive deficits associatedwith aging and Alzhei- atory amino acids. mer’s disease(Bartus et al., 1982). The basal forebrain region Rats with unilateral injections of different amounts of 192 includes the medial septum, vertical and horizontal limbs of the IgG-saporin were prepared to determine the optimal con- diagonalband of Broca, and the nucleusbasalis magnocellularis ditions in order to produce a lesion restricted to the NBM (NBM) (Mesulam et al., 1983). Basal forebrain lesionsproduce that would not destroy cholinergic afferents to hippocampus behavioral deficits in both monkeys and rats (for a review, see or nearby regions. Electroencephalographic (EEG) record- Olton and Wenk, 1987) and increasethe incidence of slow-wave ings were taken from these lesioned rats before and during activity in neocortical EEG recordings in rats (Stewart et al., treatment with scopolamine (1 mg/kg, i.p.). 1984; Buzsaki et al., 1988; Riekkinen et al., 1990a,b). These Another group of rats received bilateral NBM injections of lesionsare frequently produced by microinjection of glutamate 192 IgG-saporin and were behaviorally tested using a re- analogssuch as ibotenic (IBO) or quisqualic (QUIS) acid. Both warded, delayed-alternation task on a T-maze and a passive IBO and QUIS effectively destroy cholinergic cells within the avoidance task. Finally, histological and biochemical inves- basalforebrain, but only IBO impairs performancein behavioral tigations confirmed the effectiveness and specificity of the tasks that require learning and memory (Dunnett et al., 1987; 192 IgG-saporin. Etherington et al., 1987; Everitt et al., 1987; Markowska et al., The results showed that the 192 IgG-saporin did not de- 1990; Riekkinen et al., 1991; Wenk et al., 1992). This suggests stroy neurotensin, galanin, somatostatin, NADPH-diaphor- that the memory impairments are not related to the cholinergic ase, or neuropeptide Y neurons within the NBM. Also, biom- cell loss but may be related to the loss of noncholinergic cells arkers of cholinergic function were significantly decreased within the basal forebrain (Riekkinen et al., 1990a,b; Dunnett throughout the neocortex and within the NBM, but not in the et al., 1991; Sarter and Dudchenko, 1991; Wenk et al., 1992) olfactory bulbs, hippocampus, or dorsal caudate nucleus. such as GABA, substanceP (SP), neurotensin (NT), leu-en- lntraperitoneal injections of scopolamine, but not NBM in- kephalin (LE), somatostatin (SOM), galanin, or neuropeptideY jections of 192 IgG-saporin, increased total power across (NPY) (Candy et al., 1985). The resolution of the role of basal all frequency bands; however, slow-wave frequencies forebrain cholinergic cellsin mnemonic processeshas been ham- showed a greater increase in power as compared to fast- pered by the lack of a specific method for selectively destroying wave frequencies. Acquisition, and performance of the de- cholinergic neurons. The lack of specificity of IBO and QUIS has precluded a clear interpretation of the behavioral data. Received Nov. 29, 1993; revised Feb. 28, 1994; accepted Mar. 29, 1994. The nonspecificity of neurotoxins has also prevented a clear This work is dedicated to the memory of Dr. David S. Olton, whose guidance interpretation of the effects of basal forebrain lesionsupon neo- and inspiration made these studies possible. G.L.W. thanks Drs. Carol Barnes cortical EEG activity. Recent reports found a pronounced in- and Cindy Erickson for their valuable assistance with the EEG studies. and Ms. creasein large-amplitude, slow-wave activity predominating in Jennifer durney and Mr. Chad Livdahl for their excellent assistance in conducting the behavioral testing. This work was supported by The National Science Foun- the neocortex ipsilateral to a unilateral lesion (Stewart et al., dation, BNS 89-14941. 1984; Buzsaki et al., 1988; Riekkinen et al., 1990a,b; Vander- Correspondence should be addressed to Dr. Gary L. Wenk, Division of Neural Systems, Memory and Aging, 384 Life Sciences North, University of Arizona, wolf et al., 1993). While correlations were found between the Tucson, AZ 85724. increasein slow-waveactivity and cortical ChAT loss,the amount Copyright 0 1994 Society for Neuroscience 0270-6474/94/145986-10$05.00/O of incidental and nonspecific damage to noncholinergic fore- The Journal of Neuroscience, October 1994, f4(10) 5987 brain neurons may also underlie the changes in EEG. Therefore, struments, Tujunga, CA) with the incisor bar set 2.0 mm below the ear the use of a toxin that is specific for cholinergic neurons is bars (flat skull). The scalp was incised and retracted and holes were drilled in the skull with a dental drill. required in order to determine whether the loss of NBM cho- Unilateral lesion study. The results of these unilateral lesion studies linergic neurons is solely responsible for cortical EEG slowing. guided the choice of lesion coordinates for the behavioral studies that A powerful new approach to selective neural lesioning uses used bilateral lesions. The purpose of this study was to determine the antineuronal immunotoxins (Wiley, 1992). In the present study, optimal concentration of 192 IgG-saporin that would destroy the great- est number of NBM cholineraic cells with the least amount of cell loss basal forebrain cholinergic cells were selectively destroyed by in the horizontal limb of thediagonal band of Broca, the dorsal and injection of an antineuronal immunotoxin recently developed ventral aspects of the basal ganglia, and other surrounding structures by Wiley et al. (199 1). This compound consists of a monoclonal that may receive cholinergic innervation, for example, the olfactory antibody (192 IgG) to the low-affinity nerve growth factor (NGF’) bulbs. Our preliminary studies had already investigated the optimal receptor that has been armed with the ribosome-inactivating volume and anterior-posterior coordinates that would preclude exten- sive spread of the 192 IgG-saporin and injury to medial septal and protein saporin. Injections of this 192 IgG-saporin complex diagonal band neurons, respectively (data not shown). The unilateral produce long-lasting depletions in cholinergic markers through- lesion coordinates were as follows: 0.9 mm posterior to bregma, 2.8 out the forebrain (Book et al., 1992). The actual selectivity of mm lateral from the midline (left or right side), and 6.9 mm below the this molecule remains to be fully defined. In the only published dura for all rats. The amounts of 192 IgG-sanorin tested were 5, 10, and 22.5 ng injected in a constant total ;olume of 0.03 ~1. study, a single high-dose intracerebroventricular injection (4.69 Behavioral study. Bilateral lesion coordinates for the behavioral study pg of 192 IgG-saporin in 7.0 ~1) reduced ChAT activity through- were 0.9 mm posterior to dregma, 2.8 mm lateral from the midline, out the neocortex, hippocampus, and septal area (Nilsson et al., and 6.9 mm below the dura. The 192 IgG-saporin (10 ng in 0.03 ~1) 1992). Endogenous levels of dopamine were unaffected by in- was injected slowly (taking approximately 2 min) and the needle was jections of 192 IgG-saporin, while endogenous levels of nor- left in mace for 5 min to limit diffusion dorsal to the iniection site. This concentration of 192 IgG-saporin was chosen because the preliminary epinephrine were elevated in the hippocampus and striatum. unilateral lesion study outlined above found that this concentration The lesion significantly impaired acquisition of the Morris water provided the greatest loss of cholinergic cells that project to cortex, maze task. These data do not address the issue of which cho- without loss of hippocampal afferents, and with the most limited loss linergic neurons within the basal forebrain were responsible for of striatal cholinergic cells. Control rats had the injection cannula low- ered to the same coordinates but no injection was made. the behavioral impairments, for example, the NBM, which in- The following
Load more

Recommended publications
  • The Effect of Rosmarinic Acid on Apoptosis and Nnos Immunoreactivity Following Intrahippocampal Kainic Acid Injections in Rats
    Basic and Clinical January, February 2020, Volume 11, Number 1 Research Paper: The Effect of Rosmarinic Acid on Apoptosis and nNOS Immunoreactivity Following Intrahippocampal Kainic Acid Injections in Rats Safoura Khamse1* , Seyed Shahabeddin Sadr1,2 , Mehrdad Roghani3* , Mina Rashvand1 , Maryam Mohammadian4 , Narges Mare- fati1 , Elham Harati1 , Fatemeh Ebrahimi1 1. Department of Physiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. 2. Electrophysiology Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran. 3. Neurophysiology Research Center, Shahed University, Tehran, Iran. 4. Department of Physiology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran. Use your device to scan and read the article online Citation: Khamse, S., Sadr, Sh., Roghani, M., Rashvand, M., Mohammadian, M., & Marefati, N., et al. The Effect of Ros- marinic Acid on Apoptosis and nNOS Immunoreactivity Following Intrahippocampal Kainic Acid Injections in Rats Basic and Clinical Neuroscience, 11(1), 41-48. http://dx.doi.org/10.32598/bcn.9.10.340 http://dx.doi.org/10.32598/bcn.9.10.340 A B S T R A C T Introduction: Kainic Acid (KA) is an ionotropic glutamate receptor agonist. KA can induce neuronal overactivity and excitotoxicity. Rosmarinic Acid (RA) is a natural polyphenolic Article info: compound with antioxidant, anti-apoptotic, anti-neurodegenerative, and anti-inflammatory Received: 12 Apr 2018 properties. This study aimed to assess the effect of RA on apoptosis, nNOS-positive neurons First Revision: 10 May 2018 number, as well as Mitogen-Activated Protein Kinase (MAPK) and Cyclooxygenase-2 (COX- Accepted: 27 Oct 2018 2) immunoreactivity, following intrahippocampal Kainic acid injection in rats.
    [Show full text]
  • Kainic Acid-Induced Neurotoxicity: Targeting Glial Responses and Glia-Derived Cytokines
    388 Current Neuropharmacology, 2011, 9, 388-398 Kainic Acid-Induced Neurotoxicity: Targeting Glial Responses and Glia-Derived Cytokines Xing-Mei Zhang1 and Jie Zhu1,2, 1Department of Neurobiology, Care Sciences and Society, Karolinska Institute, Stockholm, Sweden; 2Department of Neurology, The First Hospital of Jilin University, Changchun, China Abstract: Glutamate excitotoxicity contributes to a variety of disorders in the central nervous system, which is triggered primarily by excessive Ca2+ influx arising from overstimulation of glutamate receptors, followed by disintegration of the endoplasmic reticulum (ER) membrane and ER stress, the generation and detoxification of reactive oxygen species as well as mitochondrial dysfunction, leading to neuronal apoptosis and necrosis. Kainic acid (KA), a potent agonist to the -amino- 3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate class of glutamate receptors, is 30-fold more potent in neuro- toxicity than glutamate. In rodents, KA injection resulted in recurrent seizures, behavioral changes and subsequent degeneration of selective populations of neurons in the brain, which has been widely used as a model to study the mechanisms of neurode- generative pathways induced by excitatory neurotransmitter. Microglial activation and astrocytes proliferation are the other characteristics of KA-induced neurodegeneration. The cytokines and other inflammatory molecules secreted by activated glia cells can modify the outcome of disease progression. Thus, anti-oxidant and anti-inflammatory treatment could attenuate or prevent KA-induced neurodegeneration. In this review, we summarized updated experimental data with regard to the KA-induced neurotoxicity in the brain and emphasized glial responses and glia-oriented cytokines, tumor necrosis factor-, interleukin (IL)-1, IL-12 and IL-18.
    [Show full text]
  • Hypocretin-2-Saporin Lesions of the Lateral Hypothalamus Produce Narcoleptic-Like Sleep Behavior in the Rat
    The Journal of Neuroscience, September 15, 2001, 21(18):7273–7283 Hypocretin-2-Saporin Lesions of the Lateral Hypothalamus Produce Narcoleptic-Like Sleep Behavior in the Rat Dmitry Gerashchenko,1 Matthew D. Kohls,4 MaryAnn Greco,1 Nahid S. Waleh,3 Rafael Salin-Pascual,1,2 Thomas S. Kilduff,3 Douglas A. Lappi,4 and Priyattam J. Shiromani1 1West Roxbury Veterans Affairs Medical Center and Harvard Medical School, West Roxbury, Massachusetts 02132, 2Facultad de Medicina, Universidad Nacional Auto´nomadeMe´ xico, Mexico City, Mexico 04510, 3SRI International, Menlo Park, California 94025, and 4Advanced Targeting Systems, San Diego, California 92121 Hypocretins (Hcrts) are recently discovered peptides linked to fected with the substance P (neurokinin-1) receptor. Adminis- the human sleep disorder narcolepsy. Humans with narcolepsy tration of the toxin to the LH, in which the receptor is known to have decreased numbers of Hcrt neurons and Hcrt-null mice be present, eliminated some neurons (Hcrt, melanin- also have narcoleptic symptoms. Hcrt neurons are located only concentrating hormone, and adenosine deaminase-containing in the lateral hypothalamus (LH) but neither electrolytic nor neurons) but not others (a-melanocyte-stimulating hormone), pharmacological lesions of this or any other brain region have indicating specificity of the toxin in vivo. When the toxin was produced narcoleptic-like sleep, suggesting that specific neu- administered to the LH, rats had increased slow-wave sleep, rons need to be destroyed. Hcrt neurons express the Hcrt rapid-eye movement (REM) sleep, and sleep-onset REM sleep receptor, and to facilitate lesioning these neurons, the endog- periods. These behavioral changes were negatively correlated enous ligand hypocretin-2/orexin B (Hcrt2) was conjugated to with the loss of Hcrt-containing neurons but not with the loss of the ribosome-inactivating protein saporin (SAP).
    [Show full text]
  • Advanced Targeting Systems Is Not Liable for Any Damages Resulting from the Misuse Or Handling of This Product
    ADVANCED 11175-A FLINTKOTE AVENUE, SAN DIEGO, CA 92121 TELEPHONE (858) 642-1988 • FAX (858) 642-1989 TARGETING WWW.ATSBIO.COM • [email protected] SYSTEMS mu p75-SAP (murine p75NTR-saporin) TARGETED TOXIN Catalog Number: IT-16 Quantity: 50 micrograms, 100 micrograms, 250 micrograms Format: PBS (0.14 M Sodium Chloride; 0.003 M Potassium Chloride; 0.002 M Potassium Phosphate; 0.01 M Sodium Phosphate; pH 7.4), no preservative. Sterile-filtered. Background: Targeted toxins are powerful and specific lesioning agents used in the technique known as Molecular Neurosurgery. The ribosome-inactivating protein, saporin (from the seeds of the plant, Saponaria officinalis) is bound to a targeting agent (antibody, growth factor, peptide, ligand, or cytokine). The targeted toxin is administered to the cells (in vitro or in vivo). The targeting agent seeks out and binds to its target on the cell surface. The conjugate is internalized, saporin breaks away from the targeting agent, and inactivates the ribosomes which causes protein inhibition and, ultimately, cell death. Cells which do not have the cell surface marker are not affected. Specificity and Preparation: This targeted toxin (molecular weight 210kDa) recognizes p75NTR-bearing cells in mouse. mu p75-SAP is a chemical conjugate of the rat monoclonal antibody p75NTR and the ribosome-inactivating protein, saporin (Saponaria officinalis). This product is routinely tested by cytotoxicity assay. Usage and Storage: mu p75-SAP specifically eliminates p75NTR-positive cells. Note that the concentration of this material is extremely high. This is necessary to be able to deliver, in a small volume, a sufficient amount. Due to this high concentration, the material should be handled with extreme care.
    [Show full text]
  • Strategies to Improve the Clinical Utility of Saporin-Based Targeted Toxins
    toxins Review Strategies to Improve the Clinical Utility of Saporin-Based Targeted Toxins Francesco Giansanti 1 ID , David J. Flavell 2, Francesco Angelucci 1, Maria Serena Fabbrini 3 and Rodolfo Ippoliti 1,* 1 Department of Life, Health and Environmental Sciences, University of L’Aquila, I-67100 L’Aquila, Italy; [email protected] (F.G.); [email protected] (F.A.) 2 The Simon Flavell Leukaemia Research Laboratory (Leukaemia Busters), Southampton General Hospital, Southampton, SO16 8AT, UK; [email protected] 3 Italian Ministry of Education, I-20100 Milano, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-086-243-3286 Received: 16 December 2017; Accepted: 11 February 2018; Published: 13 February 2018 Abstract: Plant Ribosome-inactivating proteins (RIPs) including the type I RIP Saporin have been used for the construction of Immunotoxins (ITxs) obtained via chemical conjugation of the toxic domain to whole antibodies or by generating genetic fusions to antibody fragments/targeting domains able to direct the chimeric toxin against a desired sub-population of cancer cells. The high enzymatic activity, stability and resistance to conjugation procedures and especially the possibility to express recombinant fusions in yeast, make Saporin a well-suited tool for anti-cancer therapy approaches. Previous clinical work on RIPs-based Immunotoxins (including Saporin) has shown that several critical issues must be taken into deeper consideration to fully exploit their therapeutic
    [Show full text]
  • Electrophysiological Mechanisms of Kainic Acid- Induced Epileptiform Activity in the Rat Hippocampal Slice’
    0270-6474/84/0405-1312$02,00/O The Journal of Neuroscience Copyright 0 Society for Neuroscience Vol. 4, No. 5, pp. 1312-1323 Printed in U.S.A. May 1984 ELECTROPHYSIOLOGICAL MECHANISMS OF KAINIC ACID- INDUCED EPILEPTIFORM ACTIVITY IN THE RAT HIPPOCAMPAL SLICE’ ROBERT S. FISHER’ AND BRADLEY E. ALGER Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland 21201 Received August 29, 1983; Revised December 28, 1983; Accepted January 4, 1984 Abstract Depression of GABA-mediated IPSPs has been proposed to be a crucial factor in the onset of epileptiform activity in most models of epilepsy. To test this idea, we studied epileptiform activity induced by bath application of the excitatory neurotoxin kainic acid (KA) in the rat hippocampal slice. Repetitive field potential firing, spontaneous or evoked, occurred during exposure to KA. Intracellular records from 52 CA1 pyramidal cells during changes from control saline to saline containing i PM KA indicated that KA depolarized cells an average of about 5 mV and caused a 15% decrease in input resistance. Action potentials and current-induced burst afterhyperpolariza- tions did not change significantly. In several cells the tonic effects of KA were preceded by a transient phase of sporadic, spontaneous depolarizations of 2 to 10 mV and 50 to 200 msec duration. These phasic depolarizations were blocked by hyperpolarization. The major effect of 1 PM KA was a depression of synaptic potentials. Initially, KA depressed fast GABA-mediated IPSPs and slow, non-GABA-mediated late hyperpolarizing potentials to 23% and 40% of control values, respectively. IPSP depression correlated closely with onset of burst potential firing in response to synaptic stimulation.
    [Show full text]
  • Neuronal Lesioning with Axonally Transported Toxins
    Journal of Neuroscience Methods 103 (2000) 73–82 www.elsevier.com/locate/jneumeth Neuronal lesioning with axonally transported toxins Ronald G. Wiley a,*, Robert H. Kline IV b a Departments of Neurology and Pharmacology, Vanderbilt Uni6ersity, VAMC Nash6ille, TN 37212-2637, USA b Laboratory of Experimental Neurology, VAMC Nash6ille, TN 37212-2637, USA Received 17 March 2000; accepted 5 July 2000 Abstract Axonally transported toxins can be used to make selective lesions of the nervous system. Collectively, these techniques are termed ‘molecular neurosurgery’ because they exploit the surface molecular identity of neurons to selectively destroy specific types of neurons. Suicide transport, is anatomically selective but not type-selective. The most widely used suicide transport agents are the toxic lectins (ricin, volkensin) and the immunotoxin, OX7-saporin. The toxic lectins and saporin are ribosome inactivating proteins that irreversibly inhibit protein synthesis. The toxic lectins have binding subunits but saporin requires a targeting vector to gain entrance into cells. Immunolesioning uses monoclonal anti-neuronal antibodies to deliver saporin selectively into neurons that express a particular target surface antigen. Neuropeptide–saporin conjugates selectively destroy neurons expressing the appropriate peptide receptors. Notable experimental uses of these agents include analysis of the function of the cholinergic basal forebrain (192-saporin) and pain research (anti-DBH-saporin, substance P-saporin). It is likely that more immunolesioning and neuropeptide-toxin conjugates will be developed in the near future. © 2000 Published by Elsevier Science B.V. Keywords: Suicide transport; Immunolesioning; Saporin; Ribosome inactivating proteins; Molecular neurosurgery Functional neuroanatomy research has long relied on been used to produce selective lesions of neurons based analysis of the effects of lesions to infer the function of on the neurotransmitters secreted, but each of these neural structures.
    [Show full text]
  • NMDA Agonists Using ALZET Osmotic Pumps
    ALZET® Bibliography References on the Administration of NMDA Agonists Using ALZET Osmotic Pumps Aspartic Acid P7453: M. Domercq, et al. Excitotoxic oligodendrocyte death and axonal damage induced by glutamate transporter inhibition. Glia 2005;52(1):36-46 ALZET Comments: Oligonucleotide, antisense; oligonucleotide sense; kainate, dihydro-; aspartic acid, DL-threo-B-benzyloxy-; Saline, sterile; CSF/CNS (optic nerve); Rabbit; 1003D; 3 days; Controls received mp w/ vehicle, or contralateral nerves; antisense (glutamate transporters GLAST + GLT-1); animal info (adult, male, white New Zealand). P6888: J. Darman, et al. Viral-induced spinal motor neuron death is non-cell-autonomous and involves glutamate excitotoxicity. Journal of Neuroscience 2004;24(34):7566-7575 ALZET Comments: Aspartic acid, dl-threo-B-hydroxy; spermine, 1-naphthyl acetyl; CSF/CNS (intrathecal, subarachnoid space); Rat; 1007D; 7 days; Controls received mp w/ saline; enzyme inhibitors (GLT-1, GluR2). P3908: A. Hirata, et al. AMPA receptor-mediated slow neuronal death in the rat spinal cord induced by long-term blockade of glutamate transporters with THA. Brain Research 1997;771(37-44 ALZET Comments: Aspartic acid, dl-threo-B-hydroxy; Glutamate, l-; CSF, artificial;; CSF/CNS (subarachnoid space, intrathecal); Rat; 2ML1; no duration posted; dose-response; cannula position verified. P0289: R. M. Mangano, et al. Chronic infusion of endogenous excitatory amino acids into rat striatum and hippocampus. Brain Res. Bull 1983;10(47-51 ALZET Comments: Aminobutyric acid, Y-; Aspartic acid, dl-threo-B-hydroxy; Aspartic acid, l-; Cysteine sulfinic acid; Glutamic acid, l-; Radio-isotopes; 3H tracer; Acetate; Saline; CSF/CNS (corpus striatum); CSF/CNS (hippocampus); Rat; 2002; 2 weeks; comparison of injec.
    [Show full text]
  • CD7 Immunotoxins in Human T-Cell Acute Lymphoblastic Leukaemia Cells
    Antibodies 2013, 2, 50-65; doi:10.3390/antib2010050 OPEN ACCESS antibodies ISSN 2073-4468 www.mdpi.com/journal/antibodies Article Dissecting the Entry Route of Saporin-based -CD7 Immunotoxins in Human T-Cell Acute Lymphoblastic Leukaemia Cells Francesco Giansanti 1, Valeria Giordani 1, Riccardo Vago 2, David J. Flavell 3, Sopsamorn U. Flavell 3, Maria Serena Fabbrini 4,* and Rodolfo Ippoliti 1,* 1 Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, I-67100, Italy 2 Dibit-HSR Scientific Institute and Università Vita-Salute San Raffaele, Milano, I-20132, Italy 3 The Simon Flavell Leukaemia Research Laboratory, Southampton General Hospital, Southampton, Hampshire SO16 6YD, UK 4 Istituto Nazionale di Genetica Molecolare-INGM, Milano, I-20122, Italy * Authors to whom correspondence should be addressed; E-Mails: [email protected] (M.S.F.); [email protected] (R.I.); Tel.: +39-086-243-3286 (R.I.); Fax: +39-086-243-3273 (R.I.). Received: 26 November 2012; in revised form: 31 December 2012 / Accepted: 7 January 2013 / Published: 16 January 2013 Abstract: Elucidating the intracellular fate(s) of targeted toxins is of fundamental importance for their optimal use as anticancer drugs, since the biochemical targets of their enzymatic activity reside in the cell cytoplasm, as in the case of the plant ribosome inactivating proteins (RIP) saporin, ricin and of bacterial toxins. In this paper, we compared the cell surface binding and cytotoxic properties of the model RIP ricin to an immunotoxin constructed with a monoclonal antibody directed against the human T-cell marker CD7 covalently linked to saporin (CD7-SAP).
    [Show full text]
  • Neurons Coexpressing Cholecystokinin-2 and Μ
    Pain Facilitatory Cells in Rostral Ventromedial Medulla: Neurons Coexpressing Cholecystokinin-2 and Mu-Opioid Receptors Item Type text; Electronic Dissertation Authors Zhang, Wenjun Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 04/10/2021 18:17:21 Link to Item http://hdl.handle.net/10150/195291 1 PAIN FACILITATORY CELLS IN ROSTRAL VENTROMEDIAL MEDULLA: NEURONS COEXPRESSING CHOLECYSTOKININ-2 AND μ-OPIOID RECEPTORS By Wenjun Zhang A Dissertation Submitted to the Faculty of the COMMITTEE ON PHARMACOLOGY AND TOXICOLOGY (GRADUATE) In Partial Fulfillment of the Requirement For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College THE UNIVERSITY OF ARIZONA 2005 2 THE UNIVERSITY OF ARIZONA GRADUATE COLLEGE As members of the Dissertation Committee, we certify that we have read the dissertation prepared by Wenjun Zhang entitled “Pain facilitatory cells in rostral ventromedial medulla: neurons coexpressing cholecystokinin-2 and mu-opioid receptors” and recommend that it be accepted as fulfilling the dissertation requirement for the Degree of Doctor of Philosophy Josephine Lai, Ph.D __________________________________________________Date: Nov. 28, 2005 Edward French, Ph.D _________________________________________________Date: Nov. 28, 2005 Frank Porreca, Ph.D___________________________________________________ Date: Nov. 28, 2005 Naomi Rance, M.D, Ph.D_____________________________________________Date: Nov. 28, 2005 Todd Vanderah, Ph.D__________________________________________________Date: Nov. 28, 2005 Final approval and acceptance of this dissertation is contingent upon the candidate’s submission of the final copies of the dissertation to the Graduate College.
    [Show full text]
  • Subconvulsive Dose of Kainic Acid Transiently Increases the Locomotor Activity of Adult Wistar Rats
    Physiol. Res. 64: 263-267, 2015 https://doi.org/10.33549/physiolres.932793 SHORT COMMUNICATION Subconvulsive Dose of Kainic Acid Transiently Increases the Locomotor Activity of Adult Wistar Rats V. RILJAK1, D. MAREŠOVÁ1, J. POKORNÝ1, K. JANDOVÁ1 1Institute of Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic Received March 28, 2014 Accepted September 1, 2014 On-line October 15, 2014 Summary Kainic acid (KA) is agonist for kainate subtype Kainic acid (KA) is a potent neurotoxic substance valuable in receptors of excitatory amino acids (Monaghan and research of temporal lobe epilepsy. We tested how subconvulsive Cotman 1982) and its administration was revealed as a dose of KA influences spontaneous behavior of adult Wistar rats. good strategy to model the clinical and neuropathological Animals were treated with 5 mg/kg of KA and tested in Laboras features of temporal lobe epilepsy (Olney et al. 1974). open field test for one hour in order to evaluate various Most commonly, a systemic administration of KA in behavioral parameters. Week after the KA treatment animals convulsive dose (10 mg/kg) is used to induce status were tested again in Laboras open field test. Finally, rat’s brains epilepticus leading to massive excitotoxic damage of were sliced and stained with Fluoro-Jade B to detect possible neuronal tissue (Doble 1999, Riljak et al. 2007). The KA neuronal degeneration. Treatment with KA increased the time induced status epilepticus is followed by latent period and spent by locomotion (p<0.01), exploratory rearing (p<0.05) and occurrence of spontaneous recurrent seizures (Albala et animals traveled longer distance (p<0.01).
    [Show full text]
  • Neuroprotective Effect of Citicoline on Retinal Cell Damage Induced by Kainic Acid in Rats
    Neuroprotective Effect of Citicoline on Retinal Cell Damage Induced by Kainic Acid in Rats Yong Seop Han, MD, In Young Chung, MD, Jong Moon Park, MD, PhD, Ji Myeong Yu, MD, PhD Department of Ophthalmology, College of Medicine, Gyeongsang National University, Jinju, Korea Purpose: To examine whether citicoline has a neuroprotective effect on kainic acid (KA)-induced retinal damage. Methods: KA (6 nmol) was injected into the vitreous of rat eyes. Citicoline (500mg/kg, i.p.) was administered to the rats once before and twice a day after KA-injection for 3- and 7-day intervals. The neuroprotective effects of citicoline were estimated by measuring the thickness of the various retinal layers using hematoxylin-eosin (H&E) staining. In addition, immunohistochemistry was conducted to elucidate the expression of endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase (nNOS). Results: Morphometric analysis of retinal damage in KA-injected eyes showed significant cell loss in the inner nuclear layer (INL) and inner plexiform layer (IPL) of the retinas at 3 and 7 days after KA injection, but not in the outer nuclear layer (ONL). At 3 days after citicoline treatment, no significant changes were detected in the retinal thickness and immunoreactivities of eNOS and nNOS. The immunoreactivities of eNOS and nNOS increased in the retina at 7 days after the KA injection. However, prolonged treatment for 7 days significantly attenuated the immunoreactivities and the reduction of thickness. Conclusions: The results indicate that citicoline
    [Show full text]