Targeting Topics 10q2

Enrichment of xenograft-competent genetically modified pig cells using a targeted toxin, isolectin BS-I-B4 conjugate

Akasaka E, Watanabe S, Himaki T, Ohtsuka M, Yoshida M, Miyoshi K, Sato M

Xenotransplantation 17(1):81-89, 2010.

Genetically-modified pigs lacking the gala1-3gal epitope may be suitable for production of organs that could be transplanted to humans. The ability to select for a homozygous population of donor somatic cells would accelerate the process of generating these animals, which would otherwise take approximately two years. The authors incubated a heterozygous population of 107 porcine embryonic fibroblasts with 1.6 µg of IB4-SAP (Cat. #IT-10). Even after six months the treated cells were negative for the agal epitope.

Severe scene learning impairment, but intact recognition memory, after cholinergic depletion of inferotemporal cortex followed by fornix transection

Browning PG, Gaffan D, Croxson PL, Baxter MG

Cereb Cortex 20(2):282-293, 2010.

In order to directly test depletion of cholinergic neurons in the inferotemporal cortex on learning and memory the authors lesioned the inferotemporal cortex, the rostral entorhinal cortex, and the perirhinal cortex of monkeys with ME20.4-SAP (Cat. #IT-15), 56-64 injections of 0.02 µg per injection). The data suggest that episodic memory is in part controlled by interactions between the fornix and cholinergic input to the inferotemporal cortex.

Unique Contributions of Distinct Cholinergic Projections to Motor Cortical Plasticity and Learning

Conner JM, Kulczycki M, Tuszynski MH

Cereb Cortex 20(11):2739-2748, 2010.

This work mapped the basal cholinergic forebrain system associations with skilled motor learning and motor function recovery after cortical injury. Rats were lesioned with 192-IgG-SAP (Cat. #IT-01). The animals received either two rostrocaudal injections of 75-112 ng; two 19-ng injections into the “prefrontal depletion site”; or two 19-ng injections into the “motor cortex depletion site.” Loss of motor cortex cholinergic systems disrupts map plasticity and skilled motor behavior, indicating that control of these systems rests within the motor cortex.

Depleting Syndecan-4+ T Lymphocytes Using Toxin-Bearing Dendritic Cell-Associated Heparan Sulfate Proteoglycan-Dependent Integrin Ligand: A New Opportunity for Treating Activated T Cell-Driven Disease

Akiyoshi H, Chung JS, Tomihari M, Cruz PD, Jr., Ariizumi K

J Immunol 184(7):3554-3561, 2010.

The dendritic cell-associated heparin sulfate proteoglycan-dependent integrin ligand (DC-HIL) exclusively associates with syndecan-4 (SD-4), which is expressed on some activated T-cells. The authors biotinylated DC-HIL and combined it with streptavidin-ZAP (Cat. #IT-27) and used in culture at a concentration of 10 µg/ml. Only activated T cells were bound and eliminated. (See cover article.)

Transplant of GABAergic precursors restores hippocampal inhibitory function in a mouse model of seizure susceptibility

Zipancic I, Calcagnotto ME, Piquer-Gil M, Mello LE, Alvarez-Dolado M

Cell Transplant 9(5):549-564, 2010.

Although medial ganglionic eminence-derived cells can be grafted into the neonatal brain and become functionally mature GABAergic neurons, it is not clear whether the grafted cells can rescue loss of function. The authors injected mice with 1.6-2.0 ng of SSP-SAP (Cat. #IT-11) into the anterior and posterior hippocampus to eliminate GABAergic interneurons. Neuron function in mice receiving the grafts returned to near normal.

Arcuate nucleus destruction does not block food deprivation-induced increases in food foraging and hoarding

Dailey MJ, Bartness TJ

Brain Res 1323:94-108, 2010.

While some aspects of food intake are understood, mechanisms that control hoarding of food have not been identified. This work investigates the role of NPY in the arcuate nucleus (Arc) in hoarding. Siberian hamsters received 48-ng injections of NPY-SAP (Cat. #IT-28) into the Arc; Blank-SAP (Cat. #IT-21) was used as a control. In lesioned animals food deprivation-induced hoarding was increased 100%, but baseline foraging and food hoarding were unchanged.

Dorsal horn neurons expressing NK-1 receptors mediate scratching in rats

Carstens EE, Carstens MI, Simons CT, Jinks SL

Neuroreport 21(4):303-308, 2010.

The itch signal is passed through the superficial dorsal horn. The authors investigated whether ablation of NK-1 receptor-expressing neurons in this area would affect itch-related scratching behavior. Rats received 20 µl of 2.27-µM SSP-SAP (Cat. #IT-11) as an intracisternal injection. The reduction in itch response to intradermal 5-hydroxytryptamine indicates that NK-1 receptor-expressing superficial dorsal horn neurons are important for spinal itch transmission.

Toxin-Coupled MHC Class I Tetramers Can Specifically Ablate Autoreactive CD8+ T Cells and Delay Diabetes in Nonobese Diabetic Mice

Vincent BG, Young EF, Buntzman AS, Stevens R, Kepler TB, Tisch RM, Frelinger JA, Hess PR

J Immunol 184(8):4196-4204, 2010.

MHC class I tetramers have been used to identify antigen-specific cells. In this work the authors used a biotinylated tetramer in conjunction with streptavidin-ZAP (Cat. #IT-27) to eliminate a specific subset of reactive T cells associated with islets in vivo. NOD mice received three 4.36-µg intravenous injections of the tetramer/saporin complex over 12 days. The onset of type I diabetes in the treated mice was significantly delayed.

An early sympathetic nervous system influence exacerbates collagen-induced arthritis via CD4+CD25+ cells

Harle P, Pongratz G, Albrecht J, Tarner IH, Straub RH

Arthritis Rheum 58(8):2347-2355, 2008.

The sympathetic nervous system can play conflicting roles in collagen-induced arthritis (CIA). CD4+CD25+ T cells can play an immunoregulatory effect in this system depending on the expression of the FoxP3 transcription factor. Mice received 5-µg ip injections of anti-DBH-SAP (Cat. #IT-03) to induce an early sympathectomy. The results indicate that the sympathetic nervous system increases disease severity in CIA by stimulating some of the proinflammatory aspects of CD4+CD25+ T cells.

An opposing time-dependent immune-modulating effect of the sympathetic nervous system conferred by altering the cytokine profile in the local lymph nodes and spleen of mice with type II collagen-induced arthritis

Harle P, Mobius D, Carr DJ, Scholmerich J, Straub RH

Arthritis Rheum 52(4):1305-1313, 2005.

In this work the authors examined the role of the sympathetic nervous system (SNS) in late stages of chronic arthritis. 5-µg intraperitoneal injections of anti-DBH-SAP (Cat. #IT-03) were given in mice. The results demonstrate that the SNS supports inflammation during the asymptomatic phase of arthritis, but inhibits inflammation during the chronic symptomatic phase.

The brainstem noradrenergic systems in stress, anxiety, and depression

Itoi K, Sugimoto N

J Neuroendocrinol 22(5):355-361, 2010.

In this review the authors examine the relationship between the central noradrenergic system, fear/anxiety states, and depression. The use of anti-DBH-SAP (Cat. #IT-03) to investigate the function of the noradrenergic system in these paradigms is described.

Evaluation of side effects through selective ablation of the mu opioid receptor expressing descending nociceptive facilitatory neurons in the rostral ventromedial medulla with dermorphin-saporin

Cao F, Chen SS, Yan XF, Xiao XP, Liu XJ, Yang SB, Xu AJ, Gao F, Yang H, Chen ZJ, Tian YK

Neurotoxicology 30(6):1096-1106, 2009.

It has been shown that injection of dermorphin-SAP (Cat. #IT-12) into the rostral ventromedial medulla (RVM) can abolish descending facilitation. In this work side effects produced by a 3-pmol injection of dermorphin-SAP into the RVM of rats were assessed (Saporin, Cat. #PR-01, was used as a control). Following select physiological functions over a three-month period post-lesion demonstrated that treatment with this targeted toxin produces no long-standing adverse toxicity.

Cardiovascular and behavioural responses to conditioned fear and restraint are not affected by retrograde lesions of A5 and C1 bulbospinal neurons

Vianna DM, Carrive P

Neuroscience 166(4):1210-1218, 2010.

To investigate the role of A5 neurons in some forms of psychological stress the authors injected 22 or 44 ng of anti-DBH-SAP (Cat. #IT-03) into the spinal cord of rats. Mouse IgG-SAP (Cat. #IT-18) was used as control. The data show that A5 presympathetic neurons are not essential for the expression of the tachycardic and pressor responses to conditioned fear and restraint.

Distinct Neural Pathways Mediate {alpha}7 Nicotinic Acetylcholine Receptor-Dependent Activation of the Forebrain

Thomsen MS, Hay-Schmidt A, Hansen HH, Mikkelsen JD

Cereb Cortex 20(9):2092-2102, 2010.

a7 nicotinic acetylcholine receptor (nAChR) agonists are potential treatments for some aspects of schizophrenia. The authors examine whether cholinergic neurons in the horizontal limb of the diagonal band of Broca (HDB) are a target for this treatment. Rats received 300-ng injections of 192-IgG-SAP (Cat. #IT-01) into the HDB. The results demonstrate that cholinergic neurons in the HDB are essential for a7 nAChR agonist activation of the medial prefrontal cortex.

Targeted Ablation of Cardiac Sympathetic Neurons Reduces the Susceptibility to Ischemia-Induced Sustained Ventricular Tachycardia in Conscious Rats

Lujan HL, Palani G, Zhang L, Dicarlo SE

Am J Physiol Heart Circ Physiol 298(5):H1330-9, 2010.

Reduction of cardiac sympathetic activity protects against ventricular tachy-arrhythmias, which are the leading cause of death in industrially-developed countries. Rats received 10-µg injections of CTB-SAP (Cat. #IT-14) into each stellate ganglion. Using comparison of ventricular tachycardia onset times after coronary artery occlusion it was found that lesioned rats were less susceptible to tachycardia events.

The hyperalgesic effects induced by the injection of angiotensin II into the caudal ventrolateral medulla are mediated by the pontine A(5) noradrenergic cell group

Marques-Lopes J, Pinho D, Albino-Teixeira A, Tavares I

Brain Res 1325:41-52, 2010.

Injection of angiotensin II into the caudal ventrolateral medulla (CVLM) has been shown to induce angiotensin type 1 receptor-mediated hyperalgesia. Here the authors lesioned the pontine A5 cell group with anti-DBH-SAP (Cat. #IT-03) to evaluate the role of these neurons in this model. Rats received a 1.1-µg injection of anti-DBH-SAP into the CVLM. Behavioral responses indicate that loss of noradrenergic neurons in the CVLM partially prevented angiotensin II-induced hyperalgesia.

Serotonin transport and metabolism in the mammary gland modulates secretory activation and involution

Marshall AM, Nommsen-Rivers LA, Hernandez LL, Dewey KG, Chantry CJ, Gregerson KA, Horseman ND

J Clin Endocrinol Metab 95(2):837-846, 2010.

This work begins to examine the role of the serotonin reuptake transporter (SERT) in the regulation of lactation homeostasis. The SERT monoclonal antibody (Cat. #AB-N09) was used for immunohistochemistry.

The cerebellum harbors a circadian oscillator involved in food anticipation

Mendoza J, Pevet P, Felder-Schmittbuhl MP, Bailly Y, Challet E

J Neurosci 30(5):1894-1904, 2010.

The authors report on a circadian oscillator in the cerebellum that is sensitive to feeding cues. Mice received icv injections of 0.12, 0.25, or 0.50 µg of OX7-SAP (Cat. #IT-02). Lesioned animals displayed attenuated food-anticipatory activity, and less locomotor activity after fasting.

Does Age Matter? Behavioral and Neuro-anatomical Effects of Neonatal and Adult Basal Forebrain Cholinergic Lesions

De Bartolo P, Cutuli D, Ricceri L, Gelfo F, Foti F, Laricchiuta D, Scattoni ML, Calamandrei G, Petrosini L

J Alzheimers Dis 20(1):207-227, 2010.

The authors characterized the differences caused by age on the effect of cholinergic lesions of the basal forebrain. Seven-day-old rats received 210-ng bilateral icv injections of 192-IgG-SAP (Cat. #IT-01). Eighty-day-old rats received 4-µg bilateral icv injections. Both experimental groups displayed similar behavior, indicating that development of a depleted cholinergic system yields similar results to cholinergic dysfunction in adulthood.

Recent Progress in Research on Ribosome Inactivating Proteins

Ng TB, Wong JH, Wang H

Curr Protein Pept Sci 11(1):37-53, 2009.

Brief descriptions of research done using 192-IgG-SAP (Cat. #IT-01), OX7-SAP (Cat. #IT-02), dermorphin-SAP (Cat. #IT-12), anti-SERT-SAP (Cat. #IT-23), SSP-SAP (Cat. #IT-11), anti-DBH-SAP (Cat. #IT-03), CTB-SAP (Cat. #IT-14), and other conjugates are provided.

Methylphenidate-induced impulsivity: pharmacological antagonism by beta-adrenoreceptor blockade

Milstein JA, Dalley JW, Robbins TW

J Psychopharmacol 24(3):309-321, 2010.

In this work bilateral 20-ng intracortical injections of anti-DBH-SAP (Cat. #IT-03) were used to examine the role of noradrenergic neurons in the control of psychostimulant-induced impulsivity. Although β-adrenoreceptor blockade abolished this impulsivity, lesioning noradrenergic neurons in the cortex had no effect.