targeted-toxins

Burjanadze M, Mataradze S, Rusadze K, Chkhikvishvili N, Dashniani M (2015) Selective lesion of GABA-ergic neurons in the medial septum by GAT1-saporin impairs spatial learning in a water-maze. Georgian Med News 240:59-64.

Summary: The authors investigated the role of GABAergic neurons in the medial septum on spatial learning using a Morris water maze test. Rats received bilateral injections totaling 162 ng of GAT-1-SAP (Cat. #IT-32) into the medial septum. Saporin (Cat. #PR-01) was used as a control. The lesioned animals displayed significant deficits during the water maze task, indicating that GABAergic neurons in the medial septum are intrinsic to organization of spatial map-driven behavior.

Related Products: GAT1-SAP (Cat. #IT-32), Saporin (Cat. #PR-01)

Nguyen H, Huppé-Gourgues F, Vaucher E (2015) Activation of the mouse primary visual cortex by medial prefrontal subregion stimulation is not mediated by cholinergic basalo-cortical projections. Front Syst Neurosci 9:1. doi: 10.3389/fnsys.2015.00001

Summary: Mice received 1 μg icv injections of mu p75-SAP (Cat. #IT-16) to eliminate NGFr-positive cells. The results indicate a link between the prelimbic and infralimbic cortices and the primary visual cortex.

Related Products: mu p75-SAP (Cat. #IT-16)

Dashniani M, Kruashvili L, Rusadze K, Matatradze S, Beselia G (2015) Effects of immunotoxic and electrolytic lesions of medial septal area on spatial short-term memory in rats. Georgian Med News 239:98-103.

Summary: In this work the authors investigated how essential septohippocampal projections are in a spatial working memory model. Rats received bilateral injections of 192-IgG-SAP (Cat. #IT-01, 600 ng total) or GAT-1-SAP (Cat. #IT-32, 195 ng total) into the medial septum. Saporin (Cat. #PR-01) was used as a control.

Related Products: 192-IgG-SAP (Cat. #IT-01), GAT1-SAP (Cat. #IT-32), Saporin (Cat. #PR-01)

Liu W, Zhang S, Gu S, Sang L, Dai C (2015) Mesenchymal stem cells recruit macrophages to alleviate experimental colitis through TGFβ1. Cell Physiol Biochem 35:858-865. doi: 10.1159/000369743

Usage: For in vivo depletion of macrophages, mice received i.v. injection of Mac-1-SAP 20 µg, twice per week.

Related Products: Mac-1-SAP mouse/human (Cat. #IT-06)

Reichel JM, Nissel S, Rogel-Salazar G, Mederer A, Käfer K, Bedenk BT, Martens H, Anders R, Grosche J, Michalski D, Härtig W, Wotjak CT (2015) Distinct behavioral consequences of short-term and prolonged GABAergic depletion in prefrontal cortex and dorsal hippocampus. Front Behav Neurosci 8:452. doi: 10.3389/fnbeh.2014.00452 PMID: 25628548

Objective: To study the effects of GABAergic interneurons on exploratory and cognitive behavior and their involvement in schizophrenia-related symptoms.

Summary: The authors demonstrated that GABAergic lesions result in distinct behavioral phenotypes specific to the target site and modulated by incubation time. They were able to simulate a pathological disease state and the consequences of prolonged GABAergic depletion.

Related Products: Anti-vGAT-SAP (Cat. #IT-71)

See Also:

• Antonucci F et al. Cracking down on inhibition: Selective removal of GABAergic interneurons from hippocampal networks. J Neurosci 32(6):1989-2001, 2012.

Kucinski A (2015) Featured Article: Impairments in gait, posture and complex movement control in rats modeling the multi-system, cholinergic-dopaminergic losses in Parkinson’s Disease. Targeting Trends 16(1)

Related Products: 192-IgG-SAP (Cat. #IT-01), Anti-ChAT-SAP (Cat. #IT-42)

Read the featured article in Targeting Trends.

See Also:

• Phillips K et al. Impairments in gait, posture and complex movement control in rats modeling the multi-system, cholinergic-dopaminergic losses in PD. Neuroscience 2014 Abstracts 692.21, 2014. Society for Neuroscience, Washington, DC

Xu M, Kobets A, Du J, Lennington J, Li L, Banasr M, Duman R, Vaccarino F, DiLeone R, Pittenger C (2015) Targeted ablation of cholinergic interneurons in the dorsolateral striatum produces behavioral manifestations of Tourette syndrome. Proc Natl Acad Sci U S A 112:893-898. doi: 10.1073/pnas.1419533112

Summary: Postmortem studies of Tourette syndrome patients has revealed a reduction in the number of specific striatal interneurons. The authors explored the hypothesis that this neuronal deficit is enough to produce the symptoms of Tourette syndrome in mice. Animals received 90-ng injections of Anti-ChAT-SAP (Cat. #IT-42) into the striatum. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The data suggest that loss of the striatal interneurons is enough to produce some, but not all, of the symptoms caused by Tourette syndrome.

Related Products: Anti-ChAT-SAP (Cat. #IT-42), Rabbit IgG-SAP (Cat. #IT-35)

Q: Our lab is getting ready to begin a project using one of your targeted toxins. We already did a preliminary experiment to try out the material, but we have a couple of questions before we start the larger project. First, do you have any protocols or references for injecting intrathecally?

A: Thank you for your inquiry. We appreciate the opportunity to get involved in projects before they begin. At Advanced Targeting Systems, we do not do any in vivo work, just in vitro, however we have collaborated with many fine laboratories that have good experience with intrathecal injections. If you search PubMed with the keywords ‘saporin’ and ‘intrathecal’ you will be able to view references that will give you good information on techniques and protocols. Prior to beginning your project you will want to submit your animal care guidelines to your IACUC committee. Turner et al. published an article that will be helpful regarding intrathecal injections. [1]

Q: The second question is in two parts: 1) how do we determine the appropriate dose, and 2) how do we know saporin is not killing indiscriminately at that dose?

A: You should always use a control when determining the appropriate dose. A basic premise of the ATS targeting technology is that if a control (saporin alone or a control conjugate) evokes a response, then the dose is too high. Whenever a new shipment of targeted toxin is received, the proper working dilution should be ascertained before beginning a project. The targeted toxin data sheet states:

“There may be lot-to-lot variation in material; working dilutions must be determined by end user. If this is a new lot, assess the proper working dilution before beginning a full experimental protocol.”

If you search on the ATS website for the species and route of administration you plan to use, you can look through the publication summaries and see the dose that was used for that particular study. That will give you a ballpark range in which to start your dose titration. Just keep in mind: if the control kills cells, the dose is too high.

References

1. Turner et al. Administration of Substances to Laboratory Animals: Routes of Administration and Factors to Consider. J Am Assoc Lab Anim Sci 50(5): 600–613, 2011.

Devesa I, Ferrándiz-Huertas C, Mathivanan S, Wolf C, Luján R, Changeux J, Ferrer-Montiel A (2014) αCGRP is essential for algesic exocytotic mobilization of TRPV1 channels in peptidergic nociceptors. Proc Natl Acad Sci U S A 111:18345-18350. doi: 10.1073/pnas.1420252111

Summary: The sensitization of transient receptor potential vanilloid 1 (TRPV1) can lead to the development and maintenance of chronic pathological pain conditions. In this work the authors determined that TRPV1 receptors use membrane insertion mechanisms in order to potentiate neuronal excitability. In order to specifically link this activity to peptidergic neurons the authors treated rat primary dorsal root ganglion cultures with 10 mM rIB4-SAP (Cat. #IT-10) to deplete the non-peptidergic neurons.

Related Products: IB4-SAP (Cat. #IT-10)

Keimpema E, Zheng K, Barde SS, Berghuis P, Dobszay MB, Schnell R, Mulder J, Luiten PG, Xu ZD, Runesson J, Langel U, Lu B, Hokfelt T, Harkany T (2014) GABAergic terminals are a source of galanin to modulate cholinergic neuron development in the neonatal forebrain. Cereb Cortex 24(12):3277-3288. doi: 10.1093/cercor/bht192

Summary: In this work the authors sought to clarify the role of galanin during brain development. Several different techniques were used including the use of Galanin-SAP (Cat. #IT-34) on primary cell cultures from the fetal forebrains of rats. Cultured basal forebrain neurons were exposed to 5 ng/ml of Galanin-SAP for 8 hours, and cell death was assessed after 72 hours. Cholinergic cells were killed by Galanin-SAP, indicating that these neurons can use extracellular galanin-2 receptors to facilitate development.

Related Products: Galanin-SAP (Cat. #IT-34)