Safety Studies Begin

Early last month (March 2001), Advanced Targeting Systems (ATS) received funding from the National Institute of Mental Health (NIMH) to begin toxicology/safety studies of Substance P-Saporin (SP-SAP), a potential therapeutic for the treatment of chronic pain. The studies will be completed under the direction of three scientists who are experts in their respective fields.

Dr. Douglas Lappi (President and Chief Scientific Officer of ATS) is principal investigator for the project and will oversee the various aspects of the studies. He is an expert in the design, construction and testing of targeted toxins. His laboratory will be producing the drug and performing quality control assays throughout the project.

Dr. Tony Yaksh (Professor of Anesthesiology and Pharmacology at the University of California, San Diego School of Medicine) will direct the administration of the drug. He is the leading expert in spinal cord delivery of experimental agents. The dog is one of the species routinely used to satisfy most regulatory requirements for drug safety evaluation. The studies will assess safety from four points: 1) intrathecal dose ranging to determine the maximum tolerated dose, 2) kinetics of cerebral spinal fluid to determine how the drug penetrates spinal tissue, is redistributed and eliminated, 3) histopathology to determine impact of drug on organs and tissue, and 4) spinal GLP safety studies to determine physiological (heart and respiratory rate, blood pressure) and behavioral (arousal, muscle tone, coordination) impacts of drug administration (4).

Dr. Patrick Mantyh (Professor, University of Minnesota, Minneapolis) has established the efficacy of SP-SAP in rats and is internationally acclaimed for his immunohistochemical analysis. His laboratory will measure parameters involving the efficacy and specificity of the SP-SAP treatment. Immunohistochemistry will help in determining where the drug travels and what impact, if any, it has on spinal cord neurons (See Figure).

spsap_dog

Figure Legend: This figure shows the staining of NeuN, CGRP and IB4 in the canine spinal cord. Immunostaining is one of the tests that will be used for determination of specificity and evaluation of bystander effects by SP-SAP. Confocal photomicrographs show the pattern of immunohistochemical labeling of the neuronal nuclear marker, NeuN, a peptidergic sensory nerve fiber marker, CGRP, and the non-peptidergic sensory nerve fiber marker, IB4, in the dog spinal cord. The NeuN staining is distributed throughout the entire gray matter, while the staining for the sensory fibers (CGRP, IB4) is localized to the dorsal horn (Photo supplied by Dr. Patrick Mantyh).

The development of SP-SAP was first published in 1997 (1) by Drs. Ronald G. Wiley and Douglas Lappi, two of the founders of ATS. Their collaboration with Dr. Patrick Mantyh led to two publications in the journal Science (2, 3). These three articles describe the results of experiments with SP-SAP in the rat.

SP-SAP is a targeted toxin that permanently eliminates cells that bear the Substance P receptor. This receptor is one of many involved in the transmission of pain signals to the brain. There are two general categories of pain to be considered in this process: 1) acute pain, a physiologically important survival tool (rose thorn pricking finger, cat claws scratching cheek), and 2) chronic or noxious pain (pain that persists beyond normal healing time), often the cause of severe pathological states. The scientists used standard models of chronic pain to determine that the perception of noxious pain in the models was greatly reduced in those animals who received injections of SP-SAP. But probably just as important, the perception of acute pain was left intact. This was an extraordinarily important finding and led ATS to the decision to begin the process of developing SP-SAP as a drug.

Over the next few months, ATS will be interacting with the Center for Drug Evaluation and Research. This organization is part of the U.S. Food and Drug Administration and will evaluate the drug development plan and make determinations about the composition and guidelines for the initial clinical trials in humans. Their preliminary feedback has been a recommendation to begin clinical trials in patients with terminal cancer whose pain is no longer treatable with opioid-based drugs such as morphine. The size of this patient population may qualify SP-SAP for development as an orphan drug.

ATS is optimistic about the therapeutic possibilities of SP-SAP. The funding from NIMH is an important first step in getting the drug development process under way. The process has already begun to obtain the additional funding needed to complete the toxicology tests required by the FDA. The present goal is to be able to begin the first clinical trials in humans before the end of 2002. Progress reports will be printed in our newsletter and on the ATS website.

References

  1. Wiley RG, Lappi DA (1997) Destruction of neurokinin-1 receptor expressing cells in vitro and in vivo using substance P-saporin. Neurosci Lett 230:97-100.
  2. Mantyh PW, Rogers S, Honore P, Allen B, Ghilardi JR, Li J, Daughters RS, Vigna SR, Lappi DA, Wiley RG, Simone DA (1997) Inhibition of hyperalgesia by ablation of lamina I spinal neurons expressing the substance P receptor. Science 278:275-279.
  3. Nichols ML, Allen BJ, Rogers SD, Ghilardi JR, Honore P, Li J, Lappi DA, Simone DA, Mantyh PW (1999) Transmission of chronic nociception by spinal neurons expressing the substance P receptor. Science 286:1558-1561.
  4. Yaksh TL, Rathbun ML, Provencher JC. (1999) Preclinical safety evaluation for spinal drugs. In: Spinal Drug Delivery, Yaksh TL (ed.), Elsevier Science B.V., Amsterdam, pp. 417-437.

Targeting Trends Newsletter 01q1

Below are links to view the quarterly newsletter Targeting Trends. If you would like to be added to the mailing list, please complete the information on our contact page.


Newsletter Highlights

  • Merging of technology makes everyone a winner (page 1-2)
  • Targeted Toxins: Safe, effective in vivo administration (page 5)
  • IB4-Saporin, Neuroscience Antibodies (page 7)

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Immunolesioning: From Spinal Cord to Brain

Dr. Ann Schreihofer, University of Virginia, contributes this issue’s article from the laboratories of ATS customers. Dr. Schreihofer summarizes her research with anti-DBH-SAP (Cat. #IT-03) to immunolesion specific rostral ventral medulla neurons that project to the spinal cord by injection of anti-DBH-SAP into the rat spinal cord. The toxin is taken up and retrogradely transported to the cell bodies, eliminates protein synthesis and causes cell death. She examines the effect of neuronal loss on sympathetic nerve activity and arterial pressure.

ATS Joins Forces with Cytometry Research

Jennifer Martin wins “ATS Abstract of the Year Award”

Recent Scientific References

Targeting Talk: In vivo Use of Targeted Toxins

  • Can you use targeted toxins in vivo?
  • How do you recommend administration of the targeted toxin ?

Targeting Ticklers (Jokes)

Targeting Teaser Winners from last issue

Targeting Tools: New Products

Targeting Technology Tutorial

Targeting Teaser (Jumble)

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Targeting Tools: IB4-SAP

IB4, or the B4 lectin isoform from Bandeiraea simplicifolia (Griffonia simplicifolia), has played an important role in the delineation of the pathways of pain transmission. One of the two major groups of primary afferents that target the spinal cord dorsal horn neurons are labeled by IB4; the other group by TrkA and peptides such as CGRP and substance P and TrkA (Snider and McMachon, Tackling pain at the source: New ideas about nociceptors. Neuron 20:629-632, 1998).

ATS has designed a reagent that can eliminate in vivo the IB4- labeled neurons by conjugating the lectin to saporin. This reagent has begun to yield important information about pain pathways.

According to work presented at the Society for Neuroscience meeting, IB4-SAP specifically eliminates the IB4-positive neurons, while sparing the peptidergic neurons (see Vulchanova et al. Role of IB4- binding sensory neurons in the effects of intradermal capsaicin injection. Soc Neurosci Mtg, New Orleans LA, 2000 Abstract #212.7 and Tarpley et al. Contribution of IB-4-positive sensory neurons to NGF-induced hyperalgesia in the rat. Soc Neurosci Mtg, New Orleans LA, 2000 Abstract #633.18).

Upon binding to the alpha- glactosyl group expressed on the cell surface, IB4-SAP becomes internalized and saporin inhibits protein synthesis, resulting in the elimination of the neurons. The cytotoxin is extremely potent, with an ED50 of 80 pM for alpha- galactosyl-expressing cells in vitro. For an excellent discussion of these two classes of primary afferents, see Basbaum AI. Distinct neurochemical features of acute and persistent pain. Proc Natl Acad Sci USA 96:7739-7743, 1999.

Targeting Talk: In Vivo Use of Targeted Toxins

Can you use targeted toxins in vivo?

Yes, Molecular Neurosurgery is designed as a tool for in vivo use.

How do you recommend administration of the targeted toxin?

There are several ways to administer the toxins depending on the cells being targeted:

  1. Direct intraparenchymal pressure microinjection can be used to deliver the targeted toxin directly to target cells. This approach has been used successfully with several toxins, including SP-Saporin (SP-SAP, Cat. #IT-07), in the striatum to kill striatal interneurons that express the NK-1 receptor. Long slow infusions (0.1 µl/min) are probably the best way to do intraparenchymal injections.
    Wiley RG and Lappi DA. Neurosci Lett 230:97-100, 1997.
  2. Targeted toxins can also be injected into terminal fields and retrogradely transported to the cell bodies. This approach has been used successfully to selectively destroy locus coeruleus noradrenergic neurons that project to the olfactory bulb by injecting anti-DBH-saporin (Anti-DBH-SAP, Cat. #IT-03) into the olfactory bulb.
    Blessing WW, Lappi DA, Wiley RG. Neurosci Lett 243:85-88, 1998.

    Intracortical injections of 192-Saporin (192-IgG-SAP, Cat. #IT-01) also have been used to destroy cholinergic basal forebrain neurons projecting to the injected patch of cortex.

    192-IgG-SAP, Cat. #IT-01) in rats can reliably destroy the cholinergic basal forebrain.
    Wiley RG, Oeltmann TN, Lappi DA. Brain Res 562:149-53, 1991.

    Lumbar subarachnoid injections of SP-Saporin (SP-SAP, Cat. #IT-07) can destroy lamina I neurons in the dorsal horn that express the NK-1 receptor.

    Mantyh PW, Rogers SD, Honore P, Allen BJ, Ghilardi JR, Li J, Daughters RS, Lappi DA, Wiley RG, Simone DA. Science 278:239-40, 1997.
  3. Lastly, SP-Saporin (SP-SAP, Cat. #IT-07) has also been applied directly to the surface of the spinal cord to kill lamina I neurons expressing NK-1 receptor. In all cases, pilot studies to determine optimal toxin dose and injection parameters are recommended.

See also: Targeted Toxins Catalog

Targeting Topics 01q1

Loss of nerve: a molecular approach to better treatment of chronic pain.

Friedrich MJ.

JAMA 283(2):187-188, 2000. PMID: 10634324

The use of SP-SAP (Cat. #IT-07) as a promising new method for chronic pain relief is discussed in this review article. Chronic pain has classically been treated in ways that frequently have adverse effects on the patient’s quality of life. Friedrich touches on recently developed toxins that are useful in techniques of molecular neurosurgery. These techniques allow the dissection of pain pathways in the brain and spinal cord which will provide not only a greater understanding of these pathways but also potential therapies for chronic pain and other pain conditions.

Mab-ZAP: a tool for evaluating antibody efficacy for use in an immunotoxin.

Kohls MD, Lappi DA.

Biotechniques 28(1):162-165, 2000. PMID: 10649788

Immunotoxins are useful tools for elimination of specific cell populations in vitro and in vivo for research and therapeutic applications. One of the factors limiting the use of immunotoxins is the selection of an appropriate antibody. Advanced Targeting Systems has created a reagent that allows researchers to select antibodies with the desired characteristics before an immuntoxin is made, purified, and assayed. Using a goat anti-murine IgG coupled to the ribosome-inactivating protein saporin (Mab-ZAP, Cat. #IT-04), researchers can screen hundreds of antibodies in a time and cost-effective manner.

Selective impairment of corticotropin-releasing factor1 (CRF1) receptor-mediated function using CRF coupled to saporin.

Maciejewski-Lenoir D, Heinrichs SC, Liu XJ, Ling N, Tucker A, Xie Q, Lappi DA, Grigoriadis DE.

Endocrinology 141(2):498-504, 2000. PMID: 10650928

Corticotropin-releasing factor 1 (CRF1) is a 41-amino acid peptide which mediates many of the body’s behavioral, autonomic, immune, and endocrine responses to stress. Reduced activation of the CRF systems plays a role in a variety of psychiatric and metabolic disease states. Maciejewski-Lenoir et al. have developed a CRF-SAP targeted toxin that can eliminate ce (Cat. #IT-13) lls expressing the CRF1 but not CRF2α receptors. These data indicate that CRF-SAP may be useful as a tool to examine receptor- selective impairment of CRF system function.

Antibody for human p75 LNTR identifies cholinergic basal forebrain of non-primate species.

Tremere LA, Pinaud R, Grosche J, Hartig W, Rasmusson DD.

Neuroreport 11(10):2177-2183, 2000. PMID: 10923666

192-SAP (Cat. #IT-01) is a highly successful reagent for eliminating cholinergic neurons in rats. Because the targeting antibody only recognizes rat p75, it is unable to be used in other species. Tremere et al. have stained basal forebrain sections with ME20.4, a monoclonal antibody to human p75 (Cat. #AB-N07) and found excellent cross- reactivity in dog, raccoon, cat, pig and rabbit. The authors state that an ME20.4- saporin conjugate could be used to produce cholinergic basal forebrain lesions in several species. Last quarter, ATS highlighted the use of ME20.4-SAP in the rabbit (Targeting Trends 1:1, 2000).

Combined lesions of cholinergic and serotonergic neurons in the rat brain using 192 IgG-saporin and 5,7-dihydroxytryptamine: neurochemical and behavioural characterization.

Lehmann O, Jeltsch H, Lehnardt O, Pain L, Lazarus C, Cassel JC.

Eur J Neurosci 12(1):67-79, 2000. PMID: 10651861

Lesioning of septohippocampal pathways has often been used as a model for Alzheimer’s disease because these lesions alter cognitive capabilities such as spatial memory. Recent work in the behavioral neurosciences has shown that other neurotransmitter systems such as GABAergic, noradrenergic, and serotonergic systems also play a role in learning and memory. Lehmann et al. combined the effects of the cholinergic immunotoxin 192-SAP (Cat. #IT-01) and the serotonergic toxin 5,7-dihydroxytryptamine to examine interactions between these two pathways. The effects of lesioning these two pathways in concert indicate that they both play roles in cognitive functions related to working memory.

Effects of cholinergic depletion on neural activity in different laminae of the rat barrel cortex.

Herron P, Schweitzer JB.

Brain Res 872(1-2):71-76, 2000. PMID: 10924677

192-SAP (Cat. #IT-01) 8.0 μg/300 g body weight, nucleus basalis of Meynert

Sustained effect of metrifonate on cerebral glucose metabolism after immunolesion of basal forebrain cholinergic neurons in rats.

Bassant MH, Poindessous-Jazat F, Schmidt BH.

Eur J Pharmacol 387(2):151-162, 2000. PMID: 10650155

192-SAP (Cat. #IT-01) 134 ng in 0.2 μl, basal forebrain

Development of a method for intraparenchymal infusions of 192 IgG-saporin: a comment on Pizzo et al. (1999).

Sarter M, Bruno JP, Miner LA, McGaughy J.

J Neurosci Methods 96(2):169-170, 2000. PMID: 10720682

Letter pertaining to use of 192-SAP (Cat. #IT-01)

Nerve growth factor (NGF) augments cortical and hippocampal cholinergic functioning after p75NGF receptor-mediated deafferentation but impairs inhibitory avoidance and induces fear-related behaviors.

Winkler J, Ramirez GA, Thal LJ, Waite JJ.

J Neurosci 20(2):834-844, 2000. PMID: 10632613

192-SAP (Cat. #IT-01) 1.0 or 2.7 μg in 10 μl, intracerebroventricular

Preserved olfactory short-term memory after combined cholinergic and serotonergic lesions using 192 IgG-saporin and 5,7-dihydroxytryptamine in rats.

Wirth S, Lehmann O, Bertrand F, Lazarus C, Jeltsch H, Cassel JC.

Neuroreport 11(2):347-350, 2000. PMID: 10674484

192-SAP (Cat. #IT-01) 2 μg, intracerebroventricular

Pain control: breaking the circuit.

Hunt SP.

Trends Pharmacol Sci 21(8):284-287, 2000. PMID: 10918627

Review and analysis of the value of SP-SAP (Cat. #IT-07) in research and as a therapeutic.

In vivo [125I]-iodobenzovesamicol binding reflects cortical cholinergic deficiency induced by specific immunolesion of rat basal forebrain cholinergic system.

Sorger D, Schliebs R, Kampfer I, Rossner S, Heinicke J, Dannenberg C, Georgi P.

Nucl Med Biol 27(1):23-31, 2000. PMID: 10755642

192-SAP (Cat. #IT-01), 2 μg into each lateral ventricle

The role of cortical cholinergic afferent projections in cognition: impact of new selective immunotoxins.

McGaughy J, Everitt BJ, Robbins TW, Sarter M.

Behav Brain Res 115(2):251-263, 2000. PMID: 11000424

Review and comparison of immunolesioning techniques using 192-SAP (Cat. #IT-01) and ME20.4-SAP (Cat. #IT-15)

Brainstem noradrenergic control of nociception is abnormal in the spontaneously hypertensive rat.

Taylor BK, Roderick RE, Basbaum AI.

Neurosci Lett 291(3):139-142, 2000. PMID: 10984626

anti-DBH-SAP (Cat. #IT-03), 5 μg

Increased susceptibility to generalized seizures after immunolesions of the basal forebrain cholinergic neurons in rats.

Silveira DC, Holmes GL, Schachter SC, Geula C, Schomer DL.

Brain Res 878(1-2):223-227, 2000. PMID: 10996157

192-SAP (Cat. #IT-01), 4 μg intracerebroventricular injection

Cover Article: Immunolesioning – From Spinal Cord to Brain

Contributed by Dr. Ann Schreihofer, University of Virginia, contributes this issue’s article from the laboratories of ATS customers. Dr. Schreihofer summarizes her research with anti-DBH-SAP (Cat. #IT-03) to immunolesion specific rostral ventral medulla neurons that project to the spinal cord by injection of anti-DBH-SAP into the rat spinal cord. The toxin is taken up and retrogradely transported to the cell bodies, eliminates protein synthesis and causes cell death. She examines the effect of neuronal loss on sympathetic nerve activity and arterial pressure.

The rostral ventrolateral medulla (RVLM) is an essential structure for the generation of the sympathetic tone that maintains arterial pressure (AP) and for the generation of many sympathetic reflexes. Spinally-projecting C1 neurons, whose firing characteristics resemble those of recorded sympathetic nerves, are located in the RVLM. These cells have been speculated to be the critical presympathetic neurons. However, the RVLM also contains non-catecholaminergic neurons whose properties suggest they may have an important role in the generation of sympathetic vasomotor tone. The relative roles of the C1 and non-C1 bulbospinal RVLM neurons have been difficult to determine. Until recently the selective depletion of either class of presympathetic neuron was not possible because the C1 cells are insensitive to classic catecholaminergic neurotoxins, such as 6-hydroxydopamine, and a marker for the non-C1 cells has not been identified. The recent development of the immunolesioning tool, saporin conjugated to an antibody for dopamine beta hydroxylase (anti-DBH-SAP), has provided an effective tool for examining the effects of the selective elimination of the catecholaminergic bulbospinal RVLM neurons.

We depleted bulbospinal C1 cells in rats by microinjection of anti-DBH-SAP bilaterally into two levels of the upper thoracic spinal cord (21 ng/100 nl/injection; 4 injections/rat). To directly examine the depletion of the bulbospinal RVLM neurons we also injected the retrograde tracer Fast Blue into two alternate levels of the spinal cord in some rats. This protocol produced an average depletion of >74% of bulbospinal C1 cells (range, 50-95%) and several other bulbospinal catecholaminergic cell groups (84% of C3 cells and 98% of A5 cells).[1,2,3] After 3-5 weeks these rats have a normal mean AP, and a sympathetic nerve activity (SNA) that continues to be modulated by baroreceptor inputs,[1,3] although the range of this reflex is reduced.[1] Stimuli that inhibit SNA and decrease AP, such as intravenous phenyl biguanide1 or clonidine,[2] appear to be unaffected by treatment with anti-DBH-SAP. In contrast, stimuli that increase SNA and AP, such as intravenous cyanide1 or electrical stimulation of the RVLM itself,[3] appear to be markedly reduced or absent after treatment with anti-DBH- SAP. These data suggest that the non-C1 bulbospinal RVLM neurons may be sufficient to maintain the basal SNA that maintains resting AP; however the C1 cells may be critical for the full expression of sympatho-excitatory responses mediated by the RVLM.

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Figure Legend: Depletion of bulbospinal C1 adrenergic neurons with intraspinal injection of anti-DBH-SAP. (A) Phenylethanolamine-N-methyltransferase-immunoreactive (PNMT-ir) neurons in the RVLM from a rat after intraspinal injection of a control toxin, saporin conjugated to a mouse IgG, showing an abundance of C1 cells. (B) Same area of section in A showing many C1 neurons retrogradely labeled from intraspinal injection of Fast Blue (Arrows). (C) PNMT-ir neurons in the RVLM from a rat treated with anti-DBH-SAP showing a massive depletion of bulbospinal C1 neurons. Arrowheads indicate C1 cells with no Fast Blue, suggesting these are not bulbospinal. (D) Same area of section in C showing neurons retrogradely labeled with Fast Blue. Asterisks indicate bulbospinal neurons that are not C1 cells, which are spared by treatment with anti-DBH- SAP. The ventral surface of the medulla is at the bottom left of each photomicrograph.

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References

  1. Schreihofer AM and Guyenet PG. Am J Physiol Regulatory Integrative Comp Physiol 279:R729-R742, 2000.
  2. Schreihofer AM and Guyenet PG. Am J Physiol Regulatory Integrative Comp Physiol 279:R1753-R1762, 2000.
  3. Schreihofer AM, Stornetta RL and Guyenet PG. J Physiol (Lond.), in press, 2000.

 

SFN Poster of the Year 2000

Awarded by ATS at Society for Neuroscience (SFN) New Orleans, LA • November 4-9, 2000

389.13 Focal Hippocampal Hyperexcitability After Focal Interneuron Ablation In The Rat By Substance P-Saporin.
J.L. Martin; R.S. Sloviter*, U. Arizona Col. of Med., Tucson, AZ, USA
featuring IT-11 SSP-SAP (Poster)

Hyperexcitability after prolonged seizures or head trauma may result from interneuron malfunction or loss; but a causal relationship is in doubt because global insults produce widespread brain damage and other effects. We have therefore sought to destroy interneurons selectively using stable Substance P-saporin (SSPsap; ATS); a neurotoxin internalized by SP receptor (SPR)-expressing neurons. Improved immunofluorescent methods revealed that most GABA-; parvalbumin (PV)-; and somatostatin (SS)-positive (+) cells of all hippocampal regions (dentate gyrus and areas CA1-CA3) are SPR+; but that granule cells; mossy cells; and CA1-3 pyramidal cells are not. Intrahippocampal injections of SSPsap or vehicle were made under urethane anesthesia in 3 sites (20nl/site) of the dorsal hippocampus of 6 male Sprague-Dawley rats/group. After 5-90 days; rats were blindly evaluated in two sites for CA1 pyramidal cell and dentate granule cell responses to perforant path stimulation (PPS). SSPsap-treated rats exhibited relatively normal responses in some sites; but pathophysiology at other sites that was virtually identical to that seen after prolonged PPS or kainate (multiple population spikes and paired-pulse disinhibition in response to 0.1-2.0Hz perforant path (PP) stimuli). Abnormal responses were observed at the earliest time tested (5 days); and at 90 days. Anatomical analysis revealed selective loss of SPR+; PV+; SS+; and GABA+ neurons; and survival of principal cells and extrinsic afferents. Importantly; "epileptic" pathophysiology was observed exclusively in areas of interneuron loss. These data indicate that the pathophysiology produced by status epilepticus or head trauma can be replicated focally by selective interneuron loss alone; and provide the first direct evidence that highly focal interneuron loss per se is capable of replicating "epileptic" disinhibition and hyperexcitability. In addition; the pathophysiology is restricted to the region of the affected interneuron somata; suggesting a highly localized influence of inhibitory interneurons. Supported by: NIH grant NS18201.

Abstracts from Society for Neuroscience (SFN) – New Orleans, LA | November 4-9, 2000

41.7 IMMUNOHISTOCHEMICAL DETECTION OF ALPHA-7 NICOTINIC RECEPTOR EXPRESSION BY TWO DISTINCT CELL TYPES IN THE DORSAL RAPHE AND LOCUS COERULEUS OF RAT.

R.S. Bitner*; A.L. Nikkel; M.W. Decker

featuring IT-03 Anti-DBH-SAP (Poster)

51.22 SENSORY TRAINING IMPROVES THE ABILITY TO PROCESS STIMULI IN BARREL CORTEX AFTER BASAL FOREBRAIN LESION.

O. Rahimi*; D. Tatham; S.L. Juliano

featuring IT-01 192-IgG-SAP (Poster)

77.13 LOCAL IMMUNOTOXIN TREATMENT PREVENTS TRANSNEURONAL LABELING OF THE INTERMEDIOLATERAL COLUMN; BUT NOT THE VENTRAL HORN; OF THE SPINAL CORD AFTER TRACER INJECTION INTO LUMBAR EPAXIAL MUSCLE.

D. Daniels1*; R.R. Miselis2;3; L.M. Flanagan-Cato1;3

featuring IT-03 Anti-DBH-SAP (Poster)

119.3 DEPLETION OF CHOLINERGIC AMACRINE CELLS DOES NOT PERTURB THE SEGREGATION OF ON AND OFF CONE BIPOLAR CELL PROJECTIONS.

E. Gunhan-Agar; P. Choudary; T.-E. Landerholm; L.-M.- Chalupa*

featuring CUSTOM (Poster)

205.8 CROSSED UNILATERAL LESIONS OF THE CHOLINERGIC BASAL FOREBRAIN (BY ME20.4IGG-SAPORIN) AND FORNIX FROM INFERIOR TEMPORAL CORTEX PRODUCE SEVERE LEARNING IMPAIRMENTS IN RHESUS MONKEYS.

A. Easton1*; R.M. Ridley2; H.F. Baker2; D. Gaffan1

featuring IT-15 ME20.4-SAP (Slide)

212.7 ROLE OF IB4-BINDING SENSORY NEURONS IN THE EFFECTS OF INTRADERMAL CAPSAICIN INJECTION.

L. Vulchanova1*; T.H. Olson1;2; R. Elde1;2; C.N. Honda1;2

featuring IT-10 IB4-SAP (Slide)

212.8 INTRATHECAL DERMORPHIN-SAPORIN DECREASES MORPHINE EFFECT IN HOTPLATE ALGESIA TESTING.

S.A. Miller1;2; D.A. Lappi3; R.G. Wiley1;2*

featuring IT-12 Dermorphin-SAP (Slide)

238.1 DISTRIBUTION OF MU-OPIOID RECEPTORS AND ACTIVATED G-PROTEINS IN RAT CINGULATE CORTEX AND ALTERATIONS FOLLOWING REMOVAL OF NORADRENERGIC AFFERENTS.

L.J. Vogt1;2*; L.J. Sim-Selley3; S.R. Childers2; R.G. Wiley4; B.A. Vogt1;2

featuring IT-03 Anti-DBH-SAP (Poster)

243.6 DERMORPHIN-SAPORIN TARGETS TONIC DESCENDING FACILITATION IN THE ROSTRAL VENTROMEDIAL MEDULLA TO BLOCK AND REVERSE NEUROPATHIC PAIN.

S.E. Burgess1; T.W. Vanderah1; P.W. Mantyh2; T.P. Malan; Jr.1; M.H. Ossipov1; D. Lappi3; J. Lai1*; F. Porreca1

featuring IT-12 Dermorphin-SAP (Poster)

243.7 CHRONIC NORADRENERGIC SPINAL DENERVATION IN RATS DOES NOT PRODUCE LONG-TERM HYPERALGESIA.

L. Jasmin1*; P. Arsenault3; P.T. Ohara2; S. Marchand3

featuring IT-03 Anti-DBH-SAP (Poster)

247.10 ALTERED OPERANT AND REFLEX RESPONSES TO NOXIOUS HEAT IN RATS WITH CENTRAL NORADRENERGIC LESIONS USING ANTIDbH-SAPORIN.

C.J. Vierck1*; P.M. Belford3; M.A. Iqbal3; C. Camara3; R.H. Kline3; D.A. Lappi2; R.G. Wiley3

featuring IT-03 Anti-DBH-SAP (Poster)

310.2 INTRATHECAL INJECTIONS OF SAPORIN CONJUGATED SUBSTANCE-P ALTER THE DEVELOPMENT OF RENAL HYPERTENSION.

J. Ciriello*; M.P. Rosas-Arellano; L.P. Solano-Flores

featuring IT-07 SP-SAP (Slide)

310.7 REGULATION OF SYMPATHETIC VASOMOTOR TONE AND ARTERIAL PRESSURE BY THE ROSTRAL VENTROLATERAL MEDULLA AFTER ELIMINATION OF C1 NEURONS IN RAT.

A.M. Schreihofer*; R.L. Stornetta; P.G. Guyenet

featuring IT-03 Anti-DBH-SAP (Slide)

354.7 DIFFERENTIAL INPUT TO A NOCICEPTIVE SPECIFIC REFLEX BY TRKA-POSITIVE AND TRKA-NEGATIVE SMALL DIAMETER DRG AFFERENTS.

J.C. Petruska1*; R.D. Johnson1;2

featuring IT-01 192-IgG-SAP (Poster)

388.16 FEEDBACK HPA AXIS TO STRESS IS IMPAIRED IN RATS WITH SELECTIVE REMOVAL OF HIPPOCAMPAL CHOLINERGIC INPUT.

J.S. Han*; J.L. Bizon; H.J. Chun; C.E. Maus; M. Gallagher

featuring IT-01 192-IgG-SAP (Poster)

389.13 FOCAL HIPPOCAMPAL HYPEREXCITABILITY AFTER FOCAL INTERNEURON ABLATION IN THE RAT BY SUBSTANCE P-SAPORIN.

J.L. Martin; R.S. Sloviter*

featuring IT-11 SSP-SAP (Poster)

502.8 ANTI-DbH-SAPORIN INJECTION INTO THE PARAVENTRICULAR NUCLEUS OF THE HYPOTHALAMUS SELECTIVELY ABOLISHES 2DG-INDUCED FEEDING WITHOUT CAUSING NONSPECIFIC TISSUE DESTRUCTION.

S. Ritter*; T.T. Dinh; K. Bugarith

featuring IT-03 Anti-DBH-SAP (Slide)

507.2 ELIMINATION OF MICROGLIA SUGGESTS THEIR INVOLVEMENT IN NEURONAL PLASTICITY.

M.M. Siddiq1,2; S.E. Tsirka2,3*

featuring IT-06 MAC-1-SAP (Poster)

516.8 SCHWANN CELLS ARE REMOVED FROM THE RAT SPINAL CORD AFTER EFFECTING RECOVERY FROM PARAPLEGIA.

G. Janni1*; T. Moallem1; D.A. Lappi3; P.T. Ohara1; L. Jasmin2

featuring IT-14 CTB-SAP (Poster)

563.1 THE BEHAVIORAL AND NEUROCHEMICAL EFFECTS OF ACUTE AND INCREMENTAL CHOLINERGIC LESIONS ON VISUAL ATTENTION DURING A 5-CHOICE SERIAL REACTION TIME TASK.

J.A. McGaughy*; J.W. Dalley; T.W. Robbins; B.J. Everitt

featuring IT-01 192-IgG-SAP (Poster)

563.13 A SPECIFIC CHOLINERGIC IMMUNOTOXIN IN MICE.

J.E. Berger-Sweeney1; S.L. Murg2; M.G. Baxter2; N.A. Stearns1; D.A. Lappi3*

featuring IT-16 mu p75-Saporin (Poster)

563.2 COMBINED CHOLINERGIC DENERVATION OF THE HIPPOCAMPUS AND POSTERIOR CINGULATE CORTEX FAILS TO IMPAIR WORKING MEMORY PERFORMANCE BUT MAY PRODUCE DEFICITS IN BEHAVIOURAL FLEXIBILITY IN THE RAT.

B.D. Winters1;2*; S.B. Dunnett1;2

featuring IT-01 192-IgG-SAP (Poster)

563.3 SELECTIVE DESTRUCTION OF BASAL FOREBRAIN CHOLINERGIC NEURONS IMPAIRS ACQUISITION OF A SPATIAL MEMORY TASK.

D.A. Johnson1*; N.J. Zambon1; R.B. Gibbs2

featuring IT-01 192-IgG-SAP (Poster)

563.4 PRESERVATION OF REACTIVITY TO SPATIAL NOVELTY IN ADULT RATS AFTER SPECIFIC BASAL FOREBRAIN 192 IGG-SAPORIN LESIONS.

L. Ricceri1; M.G. Baxter2; K.M. Frick3; J. Berger-Sweeney3

featuring IT-01 192-IgG-SAP (Poster)

563.5 A TEST OF NEGATIVE PATTERNING REVEALS SELECTIVE IMPAIRMENT IN CONFIGURAL ASSOCIATION LEARNING IN RATS WITH 192 IGG-SAPORIN LESIONS OF THE NUCLEUS BASALIS MAGNOCELLULARIS.

A.E. Butt*; K. Allen; K. Arthur; C. Cole; S. Cook; A. Gerth; M. Hoichi; C. Long; M. Noble; T. Rea; J. Rogers

featuring IT-01 192-IgG-SAP (Poster)

563.6 THE ROLE OF MEDIAL SEPTAL CHOLINERGIC AND GABAERGIC NEURONS IN SOCIAL MEMORY.

A.J. Secor1;2; J. Bishop1;2; K.C.H. Pang1;2*

featuring IT-01 192-IgG-SAP (Poster)

563.7 192 IGG-SAPORIN IMMUNOLESIONING CAUSES MARKED FACILITATION OF DOPAMINE-MEDIATED LOCOMOTOR ACTIVITY IN ADULT RATS.

A. Mattsson; S.O. Ögren; L. Olson*

featuring IT-01 192-IgG-SAP (Poster)

563.8 CONTRIBUTION OF THE CHOLINERGIC BASAL FOREBRAIN TO PROACTIVE INTERFERENCE BETWEEN STORED ODOR MEMORIES DURING ASSOCIATIVE LEARNING IN RATS: 192 IGG-SAPORIN IMMUNOTOXIC LESIONS.

E. De Rosa1*; M.E. Hasselmo2; M.G. Baxter1

featuring IT-01 192-IgG-SAP (Poster)

566.17 DBH-SAPORIN LESIONS THE LOCUS COERULEUS; BUT DOES NOT PRODUCE CATAPLEXY OR ABNORMAL REM SLEEP TRIGGERING.

C.A. Blanco-Centurion1*; R. Salin-Pascual1; D. Gerashchenko1; M.A. Greco1; D.A. Lappi2; T.S. Kilduff3; P.J. Shiromani1

featuring IT-03 Anti-DBH-SAP (Poster)

566.27 OREXIN-B CONJUGATED TO SAPORIN LESIONS LH AND TMN NEURONS AND PRODUCES NARCOLEPTIC-LIKE SLEEP IN RATS.

D. Gerashchenko1; M.-A. Greco1; R. Salin-Pascual1; T.S. Kilduff2; D.A. Lappi3; P.J. Shiromani1*

featuring IT-20 Orexin-SAP (Poster)

571.2 REDUCED ANXIETY RELATED BEHAVIOR FOLLOWING ABLATION OF AMYGDALA NEURONS EXPRESSING SUBSTANCE P RECEPTOR.

S.D. Rogers1; J.L. Salak-Johnson1;2*; M.J. Schwei1; J.D. Pomonis1; P.W. Mantyh1

featuring IT-07 SP-SAP (Poster)

633.18 CONTRIBUTION OF IB4-POSITIVE SENSORY NEURONS TO NGF-INDUCED HYPERALGESIA IN THE RAT.

J.W. Tarpley; W.J. Martin*; B.S. Baldwin; M.J. Forrest; D.E. MacIntyre

featuring IT-10 IB4-SAP (Poster)

635.13 RESPONSES OF SPINAL DORSAL HORN NEURONS TO CAPSAICIN FOLLOWING INTRATHECAL PRETREATMENT WITH SUBSTANCE P-SAPORIN TOXIN.

S.G. Khasabov1; S.D. Rogers1; P.W. Mantyh1;3; D.A. Simone1;2*

featuring IT-07 SP-SAP (Poster)

639.8 INTRACORTICAL INJECTION OF DBH-SAPORIN TARGETS NORADRENERGIC AXONS IN THE MEDIAL PREFRONTAL CORTEX OF THE RAT.

M.S. Mazei1; R.G. Wiley2; A.Y. Deutch1*

featuring IT-03 Anti-DBH-SAP (Poster)

722.6 NGF-INDUCED RAPID FUNCTIONAL PLASTICITY IN THE ADULT RAT SOMATOSENSORY CORTEX IS MEDIATED BY FIBERS ORIGINATING IN THE BASAL FOREBRAIN CHOLINERGIC SYSTEM.

N. Prakash1;2*; S.C. Cohen-Cory3; R.D. Frostig1

featuring IT-01 192-IgG-SAP (Poster)

733.4 ANATOMICAL EVIDENCE FOR GLIAL ACTIVATION AFTER INTRATHECAL LUMBOSACRAL HIV-1 GLYCOPROTEIN; GP120-INDUCED ALLODYNIA.

A. Holguin1*; C.B. Armstrong1; C.M. Twining1; E.D. Milligan1; M.K. Hansen1; M. McGorry1; K.A. O’Connor1; N. Quan2; D. Martin3; D.A. Lappi4; S.F. Maier1; L.R. Watkins1

featuring IT-06 MAC-1-SAP (Poster)

733.9 PREVENTION AND TREATMENT OF A SPONTANEOUS PAIN-LIKE BEHAVIOR FOLLOWING EXCITOTOXIC SPINAL CORD INJURY (SCI) BY ABLATION OF NEURONS EXPRESSING THE SUBSTANCE P RECEPTOR.

R.P. Yezierski1*; C.G. Yu1; R.G. Wiley2

featuring IT-11 SSP-SAP (Poster)

837.10 EFFECTS OF INTRA-BASALIS INFUSION OF D-CYCLOSERINE UPON SUSTAINED ATTENTION PERFORMANCE IN RATS.

M. Sarter*; J. Turchi

featuring IT-01 192-IgG-SAP (Poster)

837.3 NUCLEUS BASALIS MAGNOCELLULARIS AND ENRICHED HOUSING: PARTNERS IN NEURAL PATTERNS OF ATTENTION?

C. Westhead; R.K. Saari; P. Morrison; P.T. Williams; M.J. Saari*

featuring IT-01 192-IgG-SAP (Poster)

837.6 ATTENTIONAL DEMAND-RELATED ALTERATIONS IN MEDIAL PREFRONTAL NEURAL ACTIVITY OF AGED RATS DURING SUSTAINED VISUAL ATTENTION.

T.M. Gill*; M.M. Yurrita; B. Givens

featuring IT-01 192-IgG-SAP (Poster)

Targeting Trends Newsletter 00q4

Below are links to view the quarterly newsletter Targeting Trends. If you would like to be added to the mailing list, please complete the information on our contact page.


Newsletter Highlights

  • Toxicology/Safety studies with SP-SAP to begin (page 1-2)
  • Second immunotoxins: use your monoclonal or polyclonal to target cells (page 5)
  • Mouse p75 immunotoxin to be released at 2000 Society for Neuroscience (SFN) Meeting (page 7)

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CBF Lesioning in Rabbits

contributed by Dr. Thomas Beach, Sun Health Research Institute, Sun City, AZ

Product information related to cover article: ME20.4-SAP, Cat. #IT-15

Chronic Pain Therapeutic

NIDA funds ATS research on Galanin

Recent Scientific References

Targeting Talk: Second Immunotoxins

  • What is a second immunotoxin?
  • How does a second immunotoxin target?
  • What happens when the second immunotoxin gets inside the cell?
  • Are there different types of second immunotoxins available?
  • What is the ratio of antibody to second immunotoxin for in vitro testing?

Targeting Ticklers (Jokes)

Targeting Teaser Winners from last issue

Targeting Tools: New Products

Targeting Technology Tutorial

Targeting Teaser (Jumble)

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Targeting Tools: ME20.4 Monoclonal and Immunotoxin

The ME20.4 monoclonal antibody (Cat. #AB-N07) was derived from immunization of mice with WM245 melanoma cells. The antibody can be used in Western blot, immunoprecipitation, immunohistochemistry, FACS analysis, and electron microscopy. The known species reactivity includes human, primate, rabbit, raccoon, dog, cat, and sheep.

The ME20.4-SAP immunotoxin (Cat. #IT-15) is a chemical conjugation between the p75NTR monoclonal antibody and saporin, a ribosome-inactivating protein. This immunotoxin specifically eliminates p75NTR neurons in multiple species (see cover
article by Dr. Beach
).