Saturday, February 28, 2009

TRPV1 with a Twist

Kudos to Dr. Marna Erickson and her team at the University of Minnesota. They recently published excellent work demonstrating a relationship between TRPV1 -EGFR signaling and skin cancer.

Ann M. Bode, Yong-Yeon Cho, Duo Zheng, Feng Zhu, Marna E. Ericson, Wei-Ya Ma, Ke Yao, and Zigang Dong. Transient Receptor Potential Type Vanilloid 1 Suppresses Skin Carcinogenesis.Cancer Res., Feb 2009; 69: 905 - 913.

"TRPV1 interacts with EGFR, leading to EGFR degradation. Notably, the absence of TRPV1 in mice results in a striking increase in skin carcinogenesis. The TRPV1 is the first membrane receptor shown to have a tumor-suppressing effect associated with the down-regulation of another membrane receptor."

...dorsal skin samples (100 mum) were processed and immunostained. For the human skin cancer tissue array, we used anti- TRPV1 -(Neuromics), anti-EGFR (Cell Signaling), and Alexa Fluor 488 and 647-conjugated secondary antibodies. Mouse skin samples were immunostained...

Sunday, February 22, 2009

TRPV1 and Retinal Ganglion Cell Apoptosis

The publications referencing our TRPV (Vanilloid); TRPM; TRPA and TRPC Antibodies keep on coming.

In this February 2009 publication, Dr. David Calkins and his team at Vanderbilt demontrate that Retinal Ganglian Cells express the TRPV1 channel and that TRPV1 activation contributes to their death with exposure to hydrostatic pressure . We also demonstrated that activation of TRPV1 alone was sufficient to induce apoptosis of RGCs.
Rebecca M. Sappington, Tatiana Sidorova, Daniel J. Long, and David J. Calkins. TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure. Invest. Ophthalmol. Vis. Sci., Feb 2009; 50: 717 - 728.
...we used rabbit anti-mouse TRPV1 IgG (1:500; catalog number RA14113; Neuromics, Edina, MN) against the absolute C terminus of mouse TRPV1 (EDAEVFKDSMAPGEK)...

TRPV1 localization in RGCs of rat retina. (A) Immunocytochemical labeling for TRPV1 shows strong localization in the outer retina and in RGCs (large cell bodies); there is little or no label in smaller displaced amacrine cells of the GCL. Clear examples of amoeboid-shaped cell bodies of microglia cells are indicated (ovals). Right: Control section preabsorbed using the TRPV1 blocking peptide (+BP). (B) Confocal image stack through GCL and NFL showing labeling for TRPV1 in wholemount preparation counterlabeled with antibodies against heavy-chain neurofilaments that recognize broad-field RGCs (SMI32). Image shows punctate localization to dendrites (arrows) as well as intense label to cell bodies (brackets); smaller cell bodies with TRPV1 label are in the background. (C) Confocal image stack through GCL and NFL of peripheral retina shows TRPV1 in RGC cell bodies (bracket) and in small bundles of RGC axons (arrows). (D) Confocal stack from central retina shows TRPV1 in RGC cell bodies (bracket) and in axon bundles in the NFL as they course toward the optic nerve head. Amoeboid-shaped cell bodies of microglia are apparent (ovals). (E) Confocal image in single plane at GCL/NFL border shows Iba-1–labeled microglia processes colocalizing with TRPV1, as we previously demonstrated.51 TRPV1-label RGC cell bodies (brackets) and axons (arrows) are indicated for reference. (F) Immunocytochemical labeling demonstrates strong perinuclear and dendritic localization of TRPV1 in cultured RGCs counterstained with the nuclear label DAPI. Localization to dendritic processes and neurites (dashed circles) includes node-like clusters; right: these regions are shown at higher magnification. (G, top) Western blot against TRPV1 in brain and whole retina from adult rat shows band at expected molecular weight (arrowheads; 100–113 kDa). Retina demonstrates an additional band with a slightly lower molecular weight that probably corresponds to a different glycosylation state for this antibody.79 Bottom: Control Western blot with preabsorption of TRPV1 antibody using the blocking peptide prevents detection of both bands. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; NFL, nerve fiber layer.

Thursday, February 19, 2009

Neurotrophin Expression in Airway Smooth Muscle Cells

Dr. Y.K. Prakash and his team at Mayo Clinic College of Medicine have published a study of Neurotrophin Signaling in Airway Smooth Muscle (ASM) cells. Here they demonstrate that BDNF/TrkB signaling (but not BDNF/p75NTR) in human ASM cells regulates [Ca2+]i responses to agonist in both normal ASM, as well as with inflammation induced by TNFα. These effects suggest an important role of NT signaling in inflammation-induced changes in ASM contractility.

Y.S. Prakash, Michael A Thompson, and Christina M Pabelick. Brain Derived Neurotrophic Factor in TNF Modulation of Ca2+ in Human Airway Smooth Muscle. Am. J. Respir. Cell Mol. Biol., Feb 2009; doi:10.1165/rcmb.2008-0151OC.
... goat polyclonal anti-TrkB-Cat#: GT15080 (Neuromics, Minneapolis, MN; 1:1000 dilution)...

Featured Reagents
TrkB-Cat#: GT15080
TrkB-Cat#: MO15042
Related Reagents:
TrkB-Cat#: MO15025
TrkA for FC
TrkA Cat#: MO15034
TrkA Cat#: GT15153
All Neurotrophins and Growth Factor Antibodies
All Neurotrophins-Neuron/Glial Marker Antibodies

Thursday, February 12, 2009

Substance-P Pub

We re-introduced our Substance P Whole Serum Guinea Pig and Substance P Purified-Guinea Pig Antibodies in 2008.

The feedback has been positive. Related to this, here's a recent reference:

Maureen S. Riedl, Stephen A. Schnell, Aaron C. Overland, Anne-Julie Chabot-Doré, Anna M. Taylor, Alfredo Ribeiro-Da-Silva, Robert P. Elde, George L. Wilcox, Laura S. Stone.
Coexpression of 2A-adrenergic and -opioid receptors in substance P-containing terminals in rat dorsal horn. The Journal of Comparative Neurology Volume 513 Issue 4, Pages 385 - 398 Published Online: 29 Jan 2009 Copyright © 2009 Wiley-Liss, Inc., A Wiley Company.

...guinea pig anti-SP (1:500; Neuromics Antibodies)...

Related Reagents:

Substance P-Rabbit
Substance P-Mouse
Neurokinin-1 (NK 1) (RA25001)
Neurokinin-1 (NK 1) Human (RA25003)
Neurokinin-3 (NK 3) (RA25001)
proNeurokinin B (proNKB or P2)
All Neuropeptides

Image: Immuofluorescent detection of Substance P in rat spinal cord dorsal horn (red fluorescence). DAPI (blue) was used as counter stain.