Targeting Topics 14q1

BMP9 ameliorates amyloidosis and the cholinergic defect in a mouse model of Alzheimer’s disease.

Burke RM, Norman TA, Haydar TF, Slack BE, Leeman SE, Blusztajn JK, Mellott TJ.

Proc Natl Acad Sci U S A 110(48):19567-19572, 2013.

During development bone morphogenetic protein 9 (BMP9) induces the cholinergic phenotype in the basal forebrain. The authors investigated the use of BMP9 as a treatment of basal forebrain cholinergic degeneration, such as is seen in Alzheimer’s disease (AD). Transgenic mice displaying AD phenotypes and expressing GFP in cholinergic neurons received icv infusions of BMP9, and several cholinergic markers were assessed. Anti-p75NTR (Cat. #AB-N01) was used in immunoblotting at a 1:3000 dilution to measure p75 levels. The results demonstrate the protective and therapeutic activity of BMP9 on AD symptoms.

Modeling fall propensity in Parkinson’s disease: deficits in the attentional control of complex movements in rats with cortical-cholinergic and striatal-dopaminergic deafferentation.

Kucinski A, Paolone G, Bradshaw M, Albin RL, Sarter M.

J Neurosci 33(42):16522-16539, 2013.

Parkinson’s disease produces a range of symptoms, some of which are unresponsive to therapies such as levodopa. These nonmotor symptoms include cognitive impairments and deficiencies in gait and balance. Here the authors develop a system to assess fall propensity in rats and examine the interaction between loss of cortical cholinergic and striatal dopaminergic afferents. Rats received 160-ng injections of 192-IgG-SAP (Cat. #IT-01) into the nucleus basalis and substantia innominata of the basal forebrain. The results indicate that the dual lesions result in diminished striatal control of complex movement.

Intrathecal substance p-saporin in the dog: efficacy in bone cancer pain.

Brown DC, Agnello K.

Anesthesiology 119(5):1178-1185, 2013.

This work demonstrates the use of naturally occurring bone cancer in dogs as a model for pain therapy. Companion dogs with bone cancer received 20-60 μg intrathecal injections of SP-SAP (currently in human clinical trials, see cover article) depending on the size of the dog. Significantly more dogs in the control group required unblinding and adjustment of pain care than in the SP-SAP group, indicating the efficacy of SP-SAP in pain control. This study also demonstrates the validity of the dog model for testing analgesic protocols.

Intrathecal substance p-saporin in the dog: distribution, safety, and spinal neurokinin-1 receptor ablation.

Wiese AJ, Rathbun M, Butt MT, Malkmus SA, Richter PJ, Osborn KG, Xu Q, Veesart SL, Steinauer JJ, Higgins D, Lappi DA, Russell B, Yaksh TL.

Anesthesiology 119(5):1163-1177, 2013.

Here the authors investigate the safety parameters of SP-SAP on purpose-bred beagles (currently in human clinical trials, see cover article). The dogs received 1.5, 15, or 150 μg intrathecal injections of the conjugate. SP-SAP pharmacology and physiological effects were assessed by behavioral and functional observations, immunohistochemistry, ELISA, blood and urine analysis, histopathology, and in situ hybridization. The general conclusions include that neurokinin-1 receptor (NK1r) positive neuron loss is detectable as soon as 7 days after administration of SP-SAP, the neuron loss is permanent, toxicity is specific to NK1r-positive neurons, and, other than the 150 μg dose, NK1r neuron loss was restricted to the superficial dorsal horn.

Substance P-Saporin for Bone Cancer Pain in Dogs: Can Man’s Best Friend Solve the Lost in Translation Problem in Analgesic Development?

Hayashida K.

Anesthesiology 119(5):999-1000, 2013.

This editorial describes the SP-SAP papers in this latest issue of Anesthesiology (See summaries above). The results of the paper are discussed, and the potential in using companion dogs for pain models is emphasized. While most pain models have been rodent-based, companion dogs provide models for chronic pain due to natural causes such as cancer and arthritis, along with frequent opportunity for behavioral assessments by the owner. Such assessments can be done without stress to the animal.

Concordance between in vivo and postmortem measurements of cholinergic denervation in rats using PET with [18F]FEOBV and choline acetyltransferase immunochemistry.

Parent MJ, Cyr M, Aliaga A, Kostikov A, Schirrmacher E, Soucy JP, Mechawar N, Rosa-Neto P, Bedard MA.

EJNMMI Res 3(1):70, 2013.

Positron emission tomography (PET) imaging agents have been developed for the quantitative evaluation of cholinergic systems in vivo, and in this work the authors examine the concordance between the in vivo use of PET and post-mortem analysis of cholinergic damage. Rats received unilateral 0.2-0.25 μg injections of 192-IgG-SAP (Cat. #IT-01) into the nucleus basalis magnocellularis. Animals were scanned using [18F]fluoroethoxybenzovesamicol, then sacrificed for cholineacetyltransferase immunohistochemistry. The results support the use of PET as an in vivo method for analyzing the loss of cholinergic neurons.

Hindbrain Catecholamine Neurons Control Rapid Switching of Metabolic Substrate Use during Glucoprivation in Male Rats.

Li AJ, Wang Q, Dinh TT, Wiater MF, Eskelsen AK, Ritter S.

Endocrinology 154(12):4570-4579, 2013.

Previous work has shown that corticosterone secretion in response to glucoprivation is at least in part controlled by hindbrain catecholamine neurons in the paraventricular nucleus of the hypothalamus (PVH). In this work the authors investigate the metabolic consequences of lesioning these neurons. Rats received bilateral 82-ng infusions of Anti-DBH-SAP (Cat. #IT-03) into the PVH. Saporin (Cat. #PR-01) was used as a control. Although lesioned animals had the same energy expenditure and locomotor activity as controls, they also had a higher respiratory exchange ratio, indicating a reduced ability to switch from carbohydrate to fat metabolism in response to glucoprivation.

Selective lesioning of nucleus incertus with corticotropin releasing factor-saporin conjugate.

Lee LC, Rajkumar R, Dawe GS.

Brain Res 1543:179-190, 2014.

In this work the authors used CRF-SAP (Cat. #IT-13) to eliminate CRF1 receptor-expressing cells from the nucleus incertus (NI). Rats received bilateral CRF-SAP injections of 21.5 to 86 ng into the NI. Blank-SAP (Cat. #IT-21) was used as a control. Lesioned animals displayed a significant loss of CRF1 receptor-expressing cells, along with a decrease in relaxin-3 and GAD65 expression.

Selective potentiation of (alpha4)3(beta2)2 nicotinic acetylcholine receptors augments amplitudes of prefrontal acetylcholine- and nicotine-evoked glutamatergic transients in rats.

Grupe M, Paolone G, Jensen AA, Sandager-Nielsen K, Sarter M, Grunnet M.

Biochem Pharmacol 86(10):1487-1496, 2013.

Nicotinic acetylcholine receptors (nAChR) are involved in a wide range of processes in the central nervous system, many having to do with higher cognitive functions. In order to better understand how these receptors mediate attentional performance, the authors investigated glutamate release under varying conditions. In one series of experiments rats received a 160-ng injection of 192-IgG-SAP (Cat. #IT-01) into the right medial prefrontal cortex. The resulting decrease in glutamate release after the cholinergic lesion adds to the data indicating that positive modulation of nAChR may help alleviate attentional impairments caused by some brain disorders.

Hindbrain noradrenergic input to the hypothalamic PVN mediates the activation of oxytocinergic neurons induced by the satiety factor oleoylethanolamide.

Romano A, Potes CS, Tempesta B, Cassano T, Cuomo V, Lutz T, Gaetani S.

Am J Physiol Endocrinol Metab 305(10):E1266-73, 2013.

Feeding behavior and energy balance are in part controlled by signals from the gut. Oleoylethanolamide (OEA) is an acylethanolamide that is thought to play a role in this network. Since peripheral administration of OEA has effects on the nucleus of the solitary tract (NTS) and paraventricular nucleus (PVN) the authors investigated the role of noradrenergic afferent input to these areas. Rats received bilateral 84-ng injections of Anti-DBH-SAP (Cat. #IT-03) into the PVN. Mouse IgG-SAP (Cat. #IT-18) was used as a control.

Oscillatory coupling within neonatal prefrontal-hippocampal networks is independent of selective removal of GABAergic neurons in the hippocampus.

Bitzenhofer SH, Hanganu-Opatz IL.

Neuropharmacology 77C:57-67, 2013.

During cognitive tasks neuronal networks are entrained by oscillatory electrical rhythms with different frequencies. It has been proposed that GABAergic neurons in the prefrontal-hippocampal networks control this processing. The authors administered 252 ng of Anti-vGAT-SAP (Cat. #IT-71) into the ventral hippocampus of rats to examine how the GABAergic neurons could be involved. Unconjugated anti-vGAT (Cat #AB-N44) was used as a control. Hippocampal sharp waves were impaired during neonatal development, but the data indicate that oscillatory coupling between the neonatal prefrontal cortex and hippocampus is not controlled by GABAergic hippocampal interneurons.

Effects of intrathecal SNC80, a delta receptor ligand, on nociceptive threshold and dorsal horn substance p release.

Kouchek M, Takasusuki T, Terashima T, Yaksh TL, Xu Q.

J Pharmacol Exp Ther 347(2):258-264, 2013.

In this work the authors utilized three different preclinical pain models to examine the effects of intrathecal administration of the DOR agonist SNC80. One strategy was to assess NK-1 receptor (NK1r) internalization using anti-NK1r (Cat. #AB-N33AP) in IHC at a 1:3000 dilution on cryosections. The data indicate that the transmitter release from small peptidergic afferents is an effect mediated by DOR in the spinal cord.

A1 noradrenergic neurons lesions reduce natriuresis and hypertensive responses to hypernatremia in rats.

da Silva EF, Freiria-Oliveira AH, Custodio CH, Ghedini PC, Bataus LA, Colombari E, de Castro CH, Colugnati DB, Rosa DA, Cravo SL, Pedrino GR.

PLoS One 8(9):e73187, 2013.

Using bilateral 63-ng injections of Anti-DBH-SAP (Cat. #IT-03) into two levels of the caudal ventrolateral medulla, the authors assessed several pressor responses to infusion of hypertonic saline. Saporin (Cat. #PR-01) was used as a control. The results suggest that medullary noradrenergic A1 neurons are involved in the regulation of some responses to acute changes in body fluid composition.

Lesion of the commissural nucleus of the solitary tract/A2 noradrenergic neurons facilitates the activation of angiotensinergic mechanisms in response to hemorrhage.

Freiria-Oliveira AH, Blanch GT, De Paula PM, Menani JV, Colombari DS.

Neuroscience 254:196-204, 2013.

Previous work has generated conflicting data on the role of catecholaminergic A2 neurons in the nucleus of the solitary tract (NTS) in control of arterial pressure lability. The authors used Anti-DBH-SAP (Cat. #IT-03) to lesion these neurons in a hypotensive hemorrhage model. Rats received two injections of 12.6 ng into the commissural NTS. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The lesioned animals quickly recovered from hypotension, but were impaired by the icv administration of losartan.

Impaired hippocampal acetylcholine release parallels spatial memory deficits in Tg2576 mice subjected to basal forebrain cholinergic degeneration.

Laursen B, Mork A, Plath N, Kristiansen U, Frank Bastlund J.

Brain Res 1543:253-262, 2014.

The Tg2576 mouse strain provides a limited model for Alzheimer’s disease because they do not display degeneration of cholinergic neurons in the basal forebrain – the other main hallmark of Alzheimer’s disease in humans. Using 0.9 μg icv injections of mu p75-SAP (Cat. #IT-16) the authors evaluated mice that had both Aβ deposition and cholinergic depletion. The data show that these mice display cognitive decline and compromised cholinergic levels, creating a viable model for Alzheimer’s disease.

Characterization of cultured multipotent zebrafish neural crest cells.

Kinikoglu B, Kong Y, Liao EC.

Exp Biol Med (Maywood) 239(2):159-168, 2013.

This work details the isolation of neural crest cells (NCCs) from transgenic zebrafish embryos expressing GFP and flow cytometry; the authors analyzed lineage markers and differentiation of the NCCs. Anti-mu p75 (Cat. #AB-N01AP) was used in immunocytochemistry at a 1:20 dilution on fixed cells.

Selective immunotoxic lesions of basal forebrain cholinergic cells: effects on learning and memory in rats.

Baxter MG, Bucci DJ, Gorman LK, Wiley RG, Gallagher M.

Behav Neurosci 127(5):619-627, 2013.

In this reprint of a 1995 article, 192-IgG-SAP (Cat. #IT-01) was used to separate the depletion of cortical cholineacetyltransferase and behavioral impairment – which had previously been linked by research using less specific lesioning methods. Since the original 1995 publication, hundreds of papers have been published using a variety of lesioning techniques and a wide range of ATS products.

Immunoablation of cells expressing the NG2 chondroitin sulphate proteoglycan.

Leoni G, Rattray M, Fulton D, Rivera A, Butt AM.

J Anat 224(2):216-227, 2014.

In this work the authors use an antibody against the NG2-glia marker chondroitin sulphate proteoglycan (CSPG) along with Mab-ZAP (Cat. #IT-04) on cell lines and brain slices to eliminate cells expressing CSPG. The results demonstrate selective and effective killing, providing a method to study the function of these cells.

Deletion of naive T cells recognizing the minor histocompatibility antigen HY with toxin-coupled peptide-MHC class I tetramers inhibits cognate CTL responses and alters immunodominance.

Hess SM, Young EF, Miller KR, Vincent BG, Buntzman AS, Collins EJ, Frelinger JA, Hess PR.

Transpl Immunol 29(1-4):138-145, 2013.

The authors utilized biotinylated peptide-MHC class I tetramers with Streptavidin-ZAP (Cat. #IT-27) to selectively delete a specific population of alloreactive T cells in mice. Animals received iv 33-pmol injections of the toxic tetramer, and the data indicate that these toxic tetramers can prevent the induction of donor-specific responses that result in organ rejection.