saporin

Ono K, Ye Y, Viet C, Dang D, Schmidt B (2015) TRPV1 expression level in isolectin B₄-positive neurons contributes to mouse strain difference in cutaneous thermal nociceptive sensitivity. J Neurophysiol 113:3345-3355. doi: 10.1152/jn.00973.2014

Summary: In order to determine whether IB4-positive trigeminal sensory neurons affect pain sensitivity, the authors administered 2 μg of rIB4-SAP (Cat. #IT-10) to the right infraorbital foramen. Saporin (Cat. #PR-01) was used as a control.

Related Products: IB4-SAP (Cat. #IT-10), Saporin (Cat. #PR-01)

Wu Y, Song S, Liu H, Xing D, Wang X, Fei Y, Li G, Zhang C, Li Y, Zhang L (2015) Role of adrenomedullin in the cerebrospinal fluid-contacting nucleus in the modulation of immobilization stress. Neuropeptides 51:43-54. doi: 10.1016/j.npep.2015.03.007

Summary: The CSF-contacting nucleus (CSF-CN) is a brain structure containing neurons that can bidirectionally transmit signals between the brain parenchyma and the CSF. In order to better understand what regulatory peptides modulate this organ, the authors eliminated the CSF-CN of rats with a 500-ng icv injection of CTB-SAP (Cat. #IT-14). Saporin (Cat. #PR-01) was used as a control. The elimination of the CSF-CN worsened the response to chronic immobilization stress; with other data this information suggests that the CSF-CN uses adrenomedullin as a stress-related peptide.

Related Products: CTB-SAP (Cat. #IT-14), Saporin (Cat. #PR-01)

Dashniani M, Burjanadze M, Naneishvili T, Chkhikvishvili N, Beselia G, Kruashvili L, Pochkhidze N, Chighladze M (2015) Exploratory behavior and recognition memory in medial septal electrolytic, neuro- and immunotoxic lesioned rats. Physiol Res 64:755-767. doi: 10.33549/physiolres.932809

Summary: To investigate recognition memory that incorporates a spatial or temporal component, the authors lesioned the medial septum of rats using several techniques. For specific lesioning of cholinergic neurons rats received bilateral injections of 192-IgG-SAP (Cat. #IT-01, 500 ng total) into the medial septum. Saporin (Cat. #PR-01) was used as a control. While electrolytic lesions produced disruptions of spatial recognition memory, immunotoxin lesions did not, indicating that the cholinergic neurons of the septohippocampal pathway are not essential to processing this type of learning.

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

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)

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)

Holmes SE, Bachran C, Fuchs H, Weng A, Melzig MF, Flavell SU, Flavell DJ. (2015) Triterpenoid saponin augmention of saporin-based immunotoxin cytotoxicity for human leukaemia and lymphoma cells is partially immunospecific and target molecule dependent. Immunopharmacol Immunotoxicol 37(1):42-55. doi: 10.3109/08923973.2014.971964 PMID: 25347443

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.

Liu H, Yan W, Lu X, Zhang X, Wei J, Wang X, Wang T, Wu T, Cao J, Shao C, Zhou F, Zhang H, Zhang P, Zang T, Lu X, Cao J, Ding H, Zhang L (2014) Role of the cerebrospinal fluid-contacting nucleus in the descending inhibition of spinal pain transmission. Exp Neurol 261:475-485. doi: 10.1016/j.expneurol.2014.07.018

Summary: The first synapse in the pain pathway is in the spinal dorsal horn, and several sites are involved in the descending control of pain. Previous studies have suggested that cerebrospinal fluid-contacting neurons may facilitate signal transmission and substance transport between the brain parenchyma and the CSF, including processes that modulate pain transmission. The authors administered CTB-SAP (Cat. #IT-14) into the right lateral ventricle of rats. Saporin (Cat. #PR-01) was used as a control. The results indicate that the 5-HT pathway contacting the CSF is an important piece in the descending inhibitory system controlling spinal transmission of pain.

Related Products: CTB-SAP (Cat. #IT-14), Saporin (Cat. #PR-01)

Flak J, Myers B, Solomon M, McKlveen J, Krause E, Herman J (2014) Role of paraventricular nucleus-projecting norepinephrine/epinephrine neurons in acute and chronic stress. Eur J Neurosci 39:1903-1911. doi: 10.1111/ejn.12587

Summary: Chronic stress can cause dysregulation of the paraventricular nucleus (PVN) of the hypothalamus, resulting in structural and function changes in the neurons involved. There are data indicating that post-stress enhancement of norepinephrine is involved in the processing of chronic stress. In this work the authors investigated the hypothesis that PVN-projecting norepinephrine/epinephrine (NE/E) neurons are necessary for chronic stress-induced drive of the hypothalamic-pituitary-adrenocortical (HPA) axis. Rats received bilateral 8.82 ng injections of anti-DBH-SAP (Cat. #IT-03) into the PVN. Saporin (Cat. #PR-01) was used as a control. Lesioned animals displayed attenuated peak ACTH, indicating that NE/E neurons are required for ACTH release in the HPA axis during chronic stress.

Related Products: Anti-DBH-SAP (Cat. #IT-03), Saporin (Cat. #PR-01)

Li AJ, Wang Q, Dinh TT, Powers BR, Ritter S (2014) Stimulation of feeding by three different glucose-sensing mechanisms requires hindbrain catecholamine neurons. Am J Physiol Regul Integr Comp Physiol 306(4):R257-R264. doi: 10.1152/ajpregu.00451.2013

Summary: The glucoregulatory system of the brain requires catecholamine neurons in the hindbrain. he sensory mechanisms and connected circuitry controlling the response to glucose deficit are not well understood. In order to investigate different drugs that stimulate food intake but interfere with cellular glucose metabolism and transport the authors administered 82 ng of anti-DBH-SAP (Cat. #IT-03) into the paraventricular nucleus as bilateral injections. Saporin (Cat. #PR-01) was used as a control. Lesioned animals did not increase food intake in response to any of the drugs, indicating that stimulation of food intake is activated through a catecholamine-dependent pathway.

Related Products: Anti-DBH-SAP (Cat. #IT-03), Saporin (Cat. #PR-01)