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Download the Full PDF Here Oct-Nov-Dec 2015 Volume 16, Issue 4 Targeting Trends Reporting the latest news in Molecular Surgery A specific immunotoxin elucidates a causal role of striatal cholinergic system in behavioral flexibility by Sho Aoki and Jeffery R. Wickens Neurobiology Research Unit, Okinawa Institute of Science and Technology, 1919-1 Tancha, Onna, Kunigami, Okinawa 904-0495, Japan Behavioral flexibility is broadly defined as the ability to change behavioral strategy, Inside this issue: according to a change of governing rules. Accumulated evidence suggests the involvement of particular brain areas such as prefrontal cortex and striatum in this function, in which Targeting Topics Scientific References 3-5 specific brain regions play their own roles. An extension of understanding on neural substrates mediating behavioral flexibility needs a next step beyond the specificity of Targeting Talk brain regions: the specific role of different neuronal subtypes. A method utilizing specific Questions & Answers 5 neurotoxins enabled us to target and elucidate the role of neurochemically-specific Targeting Tools 7 neurons in this ability. In our recent study, 1 we demonstrated a causal role of rat Targeting Teaser cholinergic interneurons in the striatum in behavioral flexibility, using a new specific Word Quiz 8 immunotoxin targeting neurons containing choline acetyltransferase (ChAT). Comparing non-selective neuronal labeling and specific immunostaining of ChAT neurons indicated that local injections of the immunotoxin successfully and selectively damaged cholinergic neurons (Fig. 1). This result is consistent with a previous study that used Anti-ChAT-SAP Highlights to study the medial prefrontal cortex (Cat. #IT-42). 2 ◆ NEW! (pg 2) Using the selective lesion, we compared intact rats injected with saline and rats Mono-Biotin Saporin without cholinergic interneurons of either dorsomedial or ventral striatum in a set-shifting 3 ◆ Teaser Winners (pg 2) task. This task required animals to shift their attention from one stimulus dimension to ◆ Retrograde Transport Suicide Transport and Immunolesioning (pg 5) ◆ Anti-ChAT-SAP. (pg 7) NEW! PACAP-SAP Beta Product ◆ Targeting Teaser (pg 8) Denise Higgins, Editor Fig. 1: Representative coronal sections of the rat striatum show intact nuclei (NeuN) staining but clear ablation of the cholinergic interneurons with ChAT staining in lesioned cases (DMS or VS). Abbreviations; DMS: dorsomedial striatum, VS: ventral striatum, LV: lateral ventricle, CC: corpus callosum, AC, anterior commissure. Scale bar: 1 mm. Reprinted from Aoki et al. (2015). (continued on page 6) Targeting Trends , Page 2 Volume 16, Issue 4 New Product: Mono-Biotin Saporin The conjugation specialists at Advanced Targeting Now, there is Mono-Biotin Saporin , a chemical Systems are proud to announce a new addition to the conjugate between saporin and biotin. Each lot is catalog: specifically manufactured and analyzed to have an Mono-Biotin Saporin average molecular ratio of one biotin per one saporin . Cat. #BT-PR01 The assurance of consistency as well as the controlled labeling potential between lots makes this a valuable new Researchers now have more freedom in selecting research tool. Use this new tool in vitro or in vivo with conjugation strategies. Users have taken advantage of the your targeting agent to broaden your targeted toxin ability to use biotinylated targeting agents with possibilities. 1 Streptavidin-ZAP (Cat. #IT-27) a chemical conjugate between saporin and streptavidin. Streptavidin-ZAP converts biotinylated materials into targeted toxins. Saporin is a ribosome- Streptavidin is a tetrameric protein (molecular weight 53 inactivating protein, molecular kDa in its recombinant form), with each subunit able to weight 30 kDa, from seeds of bind a single biotin molecule. The bond between the plant Saponaria officinalis . streptavidin and biotin is rapid and essentially non- • Saporin is safe for reversible, unaffected by most extremes of pH, organic laboratory use under normal solvents, and denaturing reagents. It is the strongest safety conditions known noncovalent biological interaction • LD 50 in mice is 4 mg/kg (Ka = 10 15 M-1 ) between protein and ligand. The • Saporin does not have a streptavidin used to make Streptavidin-ZAP contains no method of cell entry on its carbohydrate group and has a neutral isoelectric point, own which therefore reduces the nonspecific binding as compared to avidin. A variety of molecules, including lectins, proteins, and antibodies, can be biotinylated and Reference reacted with streptavidin-labeled probes or other Minami SS, Sun B, Popat K, Kauppinen T, Pleiss M, Zhou Y, Ward ME, detection reagents for use in biological assays. Floreancig P, Mucke L, Desai T, Gan L. (2012) Selective targeting of microglia by quantum dots. J Neuroinflammation 9:22. Society for Neuroscience Experimental Biology October 17-21, 2015 April 2-6, 2016 Chicago, IL San Diego, CA Booth #662 Booth #TBA Upcoming Events Targeting Teaser Solution The solution to the puzzle was: Solve this quarter’s teaser at www.ATSbio.com/news/15q4_teaser.html Jumbles: SCLEROTOME CATHETER LUMBAR FLUID Congratulations to the INACTIVATING puzzle solvers from last What the students did when school was out for the summer. quarter. Each winner will receive a $100 Answer: They studied the SCIENCE of VACATION! ATS product credit . LAST QUARTER’S WINNERS: Seto Chice, SUNY HSC at Brooklyn * Corinne Kiessling, Binghamton University, NY Volume 16, Issue 4 Targeting Trends , Page 3 Targeting Topics: Recent Scientific Reference s Reviewed by Matthew Kohls including the mediation of positive and Alterations in the rostral ventromedial negative feedback effects of estradiol on medulla after the selective ablation of luteinizing hormone (LH). In the rat, two mu-opioid receptor expressing kisspeptin neuronal populations exist; one in neurons. the anteroventral periventricular nucleus (AVPV), and the KNDy (kisspeptin/ Harasawa I, Johansen JP, Fields HL, Porreca neurokinin B/dynorphin) neurons in the F, Meng ID. arcuate nucleus. In this work the authors Pain Epub2015. examine the role of KNDy neurons in The rostral ventromedial medulla (RVM) has estradiol positive feedback effects by both excitatory and inhibitory control over administering 10-ng bilateral injections of nociceptive neurons in the medullary dorsal Anti-EFNA4 Calicheamicin NK3-SAP (Cat. #IT-63) into the arcuate horn and spinal cord. Previous work has Conjugates Effectively Target Triple- nucleus of rats. The results indicate that demonstrated that elimination of mu-opioid Negative Breast and Ovarian Tumor- KNDy neurons use dynorphin to inhibit receptor-expressing neurons in the RVM Initiating Cells to Result in Sustained AVPV neurons, establishing a regulatory reduces stress and injury-induced behavioral Tumor Regressions. mechanism for the amplitude of steroid- induced LH surges. hypersensitivity, but the effect of losing Damelin M, Bankovich A, Park A, Aguilar J, these cells on the descending inhibitory Anderson W, Santaguida M, Aujay M, Fong S, Membrane associated cancer-oocyte system has not been examined. The authors Khandke K, Pulito V, Ernstoff E, Escarpe P, neoantigen SAS1B/ovastacin is a administered 1.2 pmol of Dermorphin-SAP Bernstein J, Pysz M, Zhong W, Upeslacis E, Lucas (Cat. #IT-12) to each side of the RVM of candidate immunotherapeutic target J, Lucas J, Nichols T, Loving K, Foord O, Hampl J, for uterine tumors. rats. Saporin (Cat. #PR-01) was used as a Stull R, Barletta F, Falahatpisheh H, Sapra P, Gerber Pires ES, D’Souza RS, Needham MA, Herr control. Characterization of RVM neurons in HP, Dylla SJ. lesioned animals showed a reduction in on- AK, Jazaeri AA, Li H, Stoler MH, Clin Cancer Res 21(18):4165-4173, 2015. and off-cells, but no change in the number of Anderson-Knapp KL, Thomas T, Mandal A, neutral cells. These data indicate that mu- Triple-negative breast cancer (TNBC) is Gougeon A, Flickinger CJ, Bruns DE, Pollok opioid receptor-expressing cells in the RVM characterized by tumors lacking HER2, BA, Herr JC. are not needed for analgesia produced by estrogen receptor, and progesterone receptor. Oncotarget Epub2015. activation of RVM neurons. TNBC has proved to be very difficult to treat, Ovastatin is a zinc matrix metallo-proteinase in large part because of the absence of CD103+ Dendritic Cells Elicit CD8+ T thought to play roles in sperm-egg consensus targets on the surface of the tumor interaction and the prevention of polyspermy Cell Responses to Accelerate Kidney cells. In this work the authors empirically in eutherians. This protein is not found in Injury in Adriamycin Nephropathy. established a set of surface markers normal adult tissues, but is expressed by Cao Q, Lu J, Li Q, Wang C, Wang XM, Lee associated with TNBC tumor initiating cells, uterine carcinosarcomas. The authors VW, Wang C, Nguyen H, Zheng G, Zhao Y, as produced by patient-derived xenografts. investigated the possibility of targeting Alexander SI, Wang Y, Harris DC. Ephrin-A4 was selected as a therapeutic ovastatin as a tumor surface neoantigen for J Am Soc Nephrol Epub2015. target, and a cell line transfected with the therapeutic purposes. SNU539 cells, a ephrin-A4 gene was challenged with two uterine malignant mixed Müllerian tumor- Although it is known that dendritic cells versions of biotinylated anti-ephrin-A4 derived cell line, were challenged with 1 (DCs) are involved in chronic kidney coupled to Streptavidin-ZAP (Cat. #IT-27). μM, 0.1 μM, and 0.01 μM rabbit polyclonal disease, it is not well understood how they Both the mouse monoclonal and the anti-ovastatin coupled to 5.42 nM Fab-ZAP either resolve or aggravate the condition. humanized antibodies reach an EC 50 of rabbit (Cat. #IT-57). Rabbit IgG-SAP (Cat. CD103+ dendritic cells in particular, are 10 ng/ml, indicating that ephrin-A4 has #IT-35) was used as a control. The results known to maintain tolerance through promise as a therapeutic target for TNBC. interaction with regulatory T cells, as well as indicate that for this form of uterine cancer, protect against infection through interactions KNDy neurons modulate the ovastatin is a viable therapeutic target.
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