Targeting Topics 14q3

Role for monocyte chemoattractant protein-1 in the induction of chronic muscle pain in the rat.

Alvarez P, Green PG, Levine JD.

Pain 155(6):1161-1167, 2014.

In order to better understand where monocyte chemoattractant protein 1 (MCP-1) fits in the chronic pain landscape the authors performed a series of experiments using antisense and mismatch oligodeoxynucleotides against the MCP-1 receptor in rats. Some animals also received 3.2 μg intrathecal injections of IB4-SAP (Cat. #IT-10). IB4-SAP treatment removed water avoidance stress-induced muscle hyperalgesia, as well as preventing stress-induced hyperalgesic priming that is a usual response to administration of MCP-1. The data indicate that MCP-1 takes action through its receptors on IB4+ nociceptors.

P2Y1 receptor-mediated potentiation of inspiratory motor output in neonatal rat in vitro.

Alvares TS, Revill AL, Huxtable AG, Lorenz CD, Funk GD.

J Physiol 592(Pt 14):3089-3111, 2014.

P2YR’s are metabotrophic purinergic receptors found in some parts of the CNS.
A subtype of this receptor excites rhythm generating networks in the preBötzinger complex. In order to better understand the role of these receptors in modulation of motor output the authors used brainstem-spinal cord and medullary slice preparations from neonatal rats to investigate P2Y1R signaling on specific neurons that innervate diaphragm and airway muscles. Anti-NK1r (Cat. #AB-N33AP) at a 1:1000 dilution was used during the immunohistochemistry. The data suggest that loss of purinergic modulation contributes to motoneuron excitability.

Orexin A activates hypoglossal motoneurons and enhances genioglossus muscle activity in rats.

Zhang GH, Liu ZL, Zhang BJ, Geng WY, Song NN, Zhou W, Cao YX, Li SQ, Huang ZL, Shen LL.

Br J Pharmacol Epub2014.

Orexin neurons are restricted to the lateral hypothalamus (LH) and are involved in functions such as feeding behavior, energy homeostasis, sleep/wake cycles, and many others. Here the authors investigate orexin control of the genioglossus – the largest upper airway dilator muscle. Rats received bilateral 172 ng injections of orexin-SAP* into the LH. Lesioned animals displayed a significant decrease in genioglossus muscle electromyograms, indicating that orexin neurons are vital to the control of this muscle.

Noradrenaline neuron degeneration contributes to motor impairments and development of L-DOPA-induced dyskinesia in a rat model of Parkinson’s disease.

Shin E, Rogers JT, Devoto P, Bjorklund A, Carta M.

Exp Neurol 257C:25-38, 2014.

Although Parkinson’s disease is usually associated with loss of dopaminergic neurons in the substantia nigra, post-mortem studies have shown that noradrenergic neurons in the locus coeruleus also degenerate. In this work the authors develop a new Parkinson’s disease model by double lesioning with both 6-OHDA into the striatum and 2.5 μg bilateral injections of anti-DBH-SAP (Cat. #IT-03) into the lateral ventricles of rats. Double-lesioned animals performed worse on tests evaluating Parkinson’s disease symptoms than those lesioned only with 6-OHDA. The data suggest that Parkinson’s disease symptoms reflect the loss of both dopaminergic and noradrenergic neurons in the midbrain.

Role of paraventricular nucleus-projecting norepinephrine/epinephrine neurons in acute and chronic stress.

Flak JN, Myers B, Solomon MB, McKlveen JM, Krause EG, Herman JP.

Eur J Neurosci 39(11):1903-1911, 2014.

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.

The cholinergic basal forebrain in the ferret and its inputs to the auditory cortex.

Bajo VM, Leach ND, Cordery PM, Nodal FR, King AJ.

Eur J Neurosci Epub2014.

The ferret has become a more common animal model in auditory neuroscience. Unlike rodent models, however, anatomical data describing the organization of the basal forebrain cholinergic system and its projections to the auditory cortex have not been well characterized. Using a variety of methods the authors mapped the architecture of the ferret basal forebrain. IHC was done with several antibodies including anti-ChAT (Cat. #AB-N34AP; 1:1000) and anti-NGFr (Cat. #AB-N07; 1:500). Animals also received 17 μg of ME20.4-SAP (Cat. #IT-15) in a total of 17 injections into the ectosylvian gyrus. The results indicate that acetylcholine is most likely involved in modulation of auditory processing.

Role of lateral hypothalamus in two aspects of attention in associative learning.

Wheeler DS, Wan S, Miller A, Angeli N, Adileh B, Hu W, Holland PC.

Eur J Neurosci Epub2014.

The lateral hypothalamic (LH) region contains both orexin and melanin-concentrating hormone (MCH) neurons. These neurons are unique to the LH but project throughout the brain. In this work the authors examined the role of the LH in specific attentional aspects of associative learning. Rats received unilateral 500 ng injections of orexin-SAP* into the LH and were tested in several learning tasks. The lesioned animals displayed impaired behavior that was correlated to the loss of orexin but not MCH neurons.

Effects of hypocretin/orexin cell transplantation on narcoleptic-like sleep behavior in rats.

Arias-Carrion O, Murillo-Rodriguez E.

PLoS One 9(4):e95342, 2014.

In this work the authors examined the effect of orexin cell grafts into the lateral hypothalamus (LH) on narcoleptic-like sleep behavior. Rats received bilateral 490-ng injections of orexin-SAP* into the LH. 21 days post-lesion, the animals then received a graft consisting of dissociated cells from a 8-10 day old rat brain. The narcoleptic-like behavior was reduced in animals receiving the graft, indicating that restoration of some orexin levels may help resolve neurodegeneration.

Effects of noradrenergic denervation by anti-DBH-saporin on behavioral responsivity to l-DOPA in the hemi-parkinsonian rat.

Ostock CY, Lindenbach D, Goldenberg AA, Kampton E, Bishop C.

Behav Brain Res 270C:75-85, 2014.

Dopamine loss is central to Parkinson’s disease and is often accompanied by noradrenergic denervation of the locus coeruleus. In this work the authors examined the role this loss plays in L-DOPA therapy using a rat Parkinson’s disease model. The rats received 10 μg of anti-DBH-SAP (Cat. #IT-03) into the left lateral ventricle. Loss of norepinephrine (NE) neurons did not affect behavior, but lesioned animals were less responsive to the pro-motor therapeutic effects of L-DOPA.

Descending controls modulate inflammatory joint pain and regulate CXC chemokine and iNOS expression in the dorsal horn.

Carr FB, Geranton SM, Hunt SP.

Mol Pain 10:39, 2014.

Peripheral joint pathology in conditions such as osteoarthritis does not always correlate to the amount of pain experienced, indicating that chronic pain is present. The role of descending facilitation in this form of chronic pain has not been investigated. The authors examined the role of mu opioid receptor-expressing cells in the rostral vental medulla (RVM) in behavioral hypersensitivity seen in joint pain models. Rats received 1.5 pmol of Dermorphin-SAP (Cat. #IT-12) into the RVM. Lesioned animals displayed prolonged attenuation of hypersensitivity, and altered expression of several genes was detected by qPCR, indicating that descending facilitation in the RVM is involved in joint pain behavior.

Cholinergic immunotoxin 192 IgG-SAPORIN alters subicular theta-gamma activity and impairs spatial learning in rats.

Rastogi S, Unni S, Sharma S, Laxmi TR, Kutty BM.

Neurobiol Learn Mem 114C:117-126, 2014.

The authors investigated the role of the subiculum in spatial informational processing, specifically cholinergic modulation of subicular theta-gamma activity. Rats received 50-ng injections of 192-IgG-SAP (Cat. #IT-01) into the ventral subiculum. Lesioned animals displayed altered theta and gamma activity as well as impaired spatial learning. The hippocampal cholinergic innervations remained intact, indicating that cholinergic modulation of theta-gamma activity in the subiculum plays an important role in spatial information processing.

Depletion of inflammatory dendritic cells with anti-CD209 conjugated to saporin toxin.

Alonso MN, Gregorio JG, Davidson MG, Gonzalez JC, Engleman EG.

Immunol Res 58(2-3):374-377, 2014.

Monocyte activity is critical in inflammatory responses, however the accumulation of inflammatory dendritic cells (DC) that are derived from monocytes can cause pathogenesis and persistence of some inflammatory diseases. The authors used a mouse model in which injection of lipopolysaccharides cause an abundance of inflammatory DC’s. The mice then received a biotinylated antibody against CD209 coupled to streptavidin-ZAP (Cat. #IT-27). Mice receiving the conjugate had lower levels of CD209-positive DC’s, indicating the potential for this strategy in reducing the DC effect during inflammatory disease.

Armodafinil-induced wakefulness in animals with ventrolateral preoptic lesions.

Vetrivelan R, Saper CB, Fuller PM.

Nat Sci Sleep 6:57-63, 2014.

Excessive daytime sleepiness is often treated with modafinil. Armodafinil, the R-isomer of modafinil, has been introduced for clinical use, but little is known about the cellular pathway targeted by these drugs. The authors examined whether armodafinil inhibits the ventrolateral preoptic nucleus (VLPO). Rats received 200-ng injections of orexin-SAP* into the VLPO, followed by administration of armodafinil. Lesioned animals displayed increased wakefulness similar to control animals, indicating that armodafinil and modafinil do not act along the VLPO neurons.

The combinational use of CRISPR/Cas9-based gene editing and targeted toxin technology enables efficient biallelic knockout of the alpha-1,3-galactosyltransferase gene in porcine embryonic fibroblasts.

Sato M, Miyoshi K, Nagao Y, Nishi Y, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S.

Xenotransplantation 21(3):291-300, 2014.

In this work the authors destroyed the function of a porcine gene involved in α-Gal production. α-Gal is involved in hyperacute rejection of pig tissues in humans. After targeting the gene, 105 cells were incubated with 0.5 μg of IB4-SAP (Cat. #IT-10) in order to eliminate any remaining α-Gal expressing cells. Surviving cells lacked α-Gal and are candidates for the creation of knockout cloned piglets.