Fragile-X, Astrocytes and BMC Image of the Month

Dr. Laurie Doering and his team at McMaster University are discovering root causes of Fragile X Syndrome. A disease manifested by cognitive impairment, attention deficit and autistic behaviours.

I wanted to share highlights and links to a recent publication as it contains interesting conclusions and some of the best multiple label staining of combined embryonic rat and mouse neurons-astrocytes cultures I have seen. No wonder that this is a highly accessed Biomed Central Article and includes the image of the month. The featured  image references use of our MAP-2 antibody.

Shelley Jacobs , Meera Nathwani and Laurie C Doering. Fragile X astrocytes induce developmental delays in dendrite maturation and synaptic protein expression. BMC Neuroscience 2010, 11:132doi:10.1186/1471-2202-11-132.

Conclusions: These experiments are the first to establish a role for astrocytes in the delayed growth characteristics and abnormal morphological features in dendrites and synapses that characterize the Fragile X syndrome.

Image: Co-culture of embryonic mouse hippocampal neurons and astrocytes. Primary embryonic hippocampal neurons at 7 days in vitro, were stained with Microtubule Associated Protein-2 (MAP, green) to enable the visualization of the dendritic arbors. These neurons were cultured on top of a monolayer of primary cortical astrocytes, stained with an antibody directed against Glial Fibrillary Acidic Protein (GFAP, red). The cell nuclei were visualized by staining with 4',6-diamidino-2-phenylindole (DAPI, blue).

Related Links:
Neuronal-Glial Markers-Astrocytes, Glia, Microglia, Olidogodendrocytes, Progenitors and Schwann Cell Markers
Neurofilament or NF Antibodies
Stem Cell Research Antibodies
Stem Cell Research Reagents

Primary Neurons and Astrocytes-Primary human, rat and mouse neurons and astrocytes.

Cancer-Induced Bone Pain

Bone crushing pain. This describes pain of the highest order. Our friend, Dr. Joseph Ghilardi, VAMC-Mpls. and his colleague, Dr. Patrick Manthy are finding the root causes of the intense and growing pain suffered by Cancer Victims. Here are highlights of a recent study:

Pain frequently accompanies cancer. What remains unclear is why this pain frequently becomes more severe and difficult to control with disease progression. Here we test the hypothesis that with disease progression, sensory nerve fibers that innervate the tumor-bearing tissue undergo a pathological sprouting and reorganization, which in other nonmalignant pathologies has been shown to generate and maintain chronic pain. Injection of canine prostate cancer cells into mouse bone induces a remarkable sprouting of calcitonin gene-related peptide (CGRP+) and neurofilament 200 kDa (NF200+) sensory nerve fibers. Nearly all sensory nerve fibers that undergo sprouting also coexpress tropomyosin receptor kinase A (TrkA+). This ectopic sprouting occurs in sensory nerve fibers that are in close proximity to colonies of prostate cancer cells, tumor-associated stromal cells and newly formed woven bone, which together form sclerotic lesions that closely mirror the osteoblastic bone lesions induced by metastatic prostate tumors in humans. Preventive treatment with an antibody that sequesters nerve growth factor (NGF), administered when the pain and bone remodeling were first observed, blocks this ectopic sprouting and attenuates cancer pain. Interestingly, reverse transcription PCR analysis indicated that the prostate cancer cells themselves do not express detectable levels of mRNA coding for NGF. This suggests that the tumor-associated stromal cells express and release NGF, which drives the pathological reorganization of nearby TrkA+ sensory nerve fibers. Therapies that prevent this reorganization of sensory nerve fibers may provide insight into the evolving mechanisms that drive cancer pain and lead to more effective control of this chronic pain state.

Image: Image:Shows rat mixed neuron/glial cultures stained with mouse monoclonal antibody to neurofilament subunit NF-L clone 7D1 (green) and chicken antibody to neurofilament NF-H. This antibody binds primarily to the phosphorylated axonal forms of NF-H, in contrast to the NF-L antibody which stains both axonal and dendritic/perikaryal neurofilaments. The NF-L antibody therefore reveals a prominent cell body in green, while the surrounding axonal profiles are orange, since the are bound by both NF-L and the chicken NF-H antibody. Blue is a DNA stain. Protocol on data sheet.

 Juan M. Jimenez-Andrade, Aaron P. Bloom, James I. Stake, William G. Mantyh, Reid N. Taylor, Katie T. Freeman, Joseph R. Ghilardi, Michael A. Kuskowski, and Patrick W. Mantyh Pathological Sprouting of Adult Nociceptors in Chronic Prostate Cancer-Induced Bone Pain. J. Neurosci., Nov 2010; 30: 14649 - 14656 ; doi:10.1523/JNEUROSCI.3300-10.2010
Here're several other pubs referencing use of our antibodies in studying bone cancer pain:

Kyle G. Halvorson, BA, Molly A. Sevcik, BA, Joseph R. Ghilardi, BS, BA, Lucy J. Sullivan, BA, Nathan J. Koewler, BS, Frieder Bauss, PhD, and Patrick W. Mantyh, PhD. Intravenous Ibandronate Rapidly Reduces Pain, Neurochemical Indices of Central Sensitization, Tumor Burden, and Skeletal Destruction in a Mouse Model of Bone Cancer. Published online 2008 April 14. doi: 10.1016/j.jpainsymman.2007.10.005
...pro-dynorphin (DYN, polyclonal guinea pig anti-rat, 1:1,000; Neuromics, Minneapolis, MN)...

Timothy K. Y. Kaan, Ping K. Yip, Sital Patel, Meirion Davies, Fabien Marchand, Debra A. Cockayne, Philip A. Nunn, Anthony H. Dickenson, Anthony P. D. W. Ford, Yu Zhong, Marzia Malcangio, and Stephen B. McMahon Systemic blockade of P2X3 and P2X2/3 receptors attenuates bone cancer pain behaviour in rats. Brain, September 2010; 133: 2549 - 2564.
......Slides were then incubated with rabbit anti-P2X3 (1:2000, Neuromics) and sheep anti-calcitonin gene-related peptide (1:1000, Biomol...anti-beta-III-tubulin (1:4000, Promega) and guinea pig anti-P2X3 (1:100, Neuromics). The next day, after three washes with phosphate-buffered......

I will keep you posted on this important topic.

Our TRPV1 Antibodies Rock

We now have 40 publications referencing use of our TRP Antibodies in multiple applications. Here are the October-November 2010 publications referencing use of these antibodies:

VR1 N-Terminus (TRPV1)-Rabbit

T. Wu, L. Song, X. Shi, Z. Jiang, J. Santos-Sacchi and A.L. Nuttal. Effect of capsaicin on potassium conductance and electromotility of guinea pig outer hair cell. doi:10.1016/j.heares.2010.10.010
...anti-TRPV1 (rabbit polyclonal, RA10110, Neuromics, Edina, MN, USA) diluted to 1:500 with 1% BSA-PBS...antibody (TRPV-1) and its blocking peptide (104 M) (Neuromics, Edina, MN, USA)...

(TRPV1) - mouse specific
Julie A. Christianson, Klaus Bielefeldt, Sacha A. Malin and Brian M. Davis. Neonatal colon insult alters growth factor expression and TRPA1 responses in adult mice. Pain Volume 151, Issue 2, November 2010, Pages 540-549.
...primary antiserum to TRPV1 (1:4000; Neuromics, Minneapolis, MN; cat# RA14113)...

VR1 C-terminus (TRPV1)-Guinea Pig
N. Schuelert, C. Zhang, A.J. Mogg, L.M. Broad, D.L. Hepburn, E.S. Nisenbaum, M.P. Johnson and J.J. McDougal. Paradoxical effects of the cannabinoid CB2 receptor agonist GW405833 on rat osteoarthritic knee joint pain. Osteoarthritis and Cartilage Volume 18, Issue 11, November 2010, Pages 1536-1543.
...primary antiserum to TRPV1 (1:4000; Neuromics, Minneapolis, MN; cat# RA14113)...

Mariusz Mucha, Lezanne Ooi, John E. Linley, Pawel Mordaka, Carine Dalle, Brian Robertson, Nikita Gamper, and Ian C. Wood Transcriptional Control of KCNQ Channel Genes and the Regulation of Neuronal Excitability.
J. Neurosci., Oct 2010; 30: 13235 - 13245 ; doi:10.1523/JNEUROSCI.1981-10.2010
...1:1000 guinea pig anti-TRPV1 (Neuromics)...

Related Reagents:
All TRP Antibodies
Pain and Inflammation Research Antibodies
Neurotransmission -Neurotransmission Research Antibody Categories