Cells from Diseased Donors

Focus on Neuro Diseases

We now have the capability to provide cells of the central and peripheral nervous system from donors with Neuro diseases. These include cells from donors with ALS, AD, PD, and Brain Cancer Donors, to name a few.

We have provided cells to virtually all the large Pharmas and many small and mid-size Bio-techs. We have worked with Novartis to gain 21-CFR compliance for cells that they are using for their eye diseases drug discovery programs.

I am at your “beck and call” should you have interested in exploring specific capabilities further. You can e-mail or call me at 612-801-1007

Parkinson's Disease and the Striatum

Excellent Video
Researchers at the Karolinska Institute recently released a video outlining their findings regarding the root causes of Parkinson's.

Highlights

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Striatal sensory responses were studied by whole-cell recordings and optogenetics

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Dopamine (DA) depletion affects intrinsic and sensory properties in direct pathway neurons

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The encoding of bilateral tactile stimuli is impaired following DA depletion

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Administration of L-DOPA can correct sensory deficits caused by DA depletion.

for more see: https://doi.org/10.1016/j.neuron.2017.05.004

This study came to my attention the researchers used our Enkaphalin Antibody as a marker for the Dopaminergic Neurons. 

Image: Mouse striatum stained with D2 cell marker Enkephalin (RA14124) in green and with neuronal marker NeuN in red courtesy of Dr Heike Rebholz of City College of New York

Leptin and Remyelination

Leptin Promotes Proliferation of OPCs

Demyelination occurs in many diseases of the Central Nervous System (CNS) including Multiple Sclerosis and Parkinson's Disease. Here researchers show that Leptin plays a role in the proliferation of Oligodendrocyte Precursor Cells (OPCs): These cells are critical for keeping the myelin sheath on Neurons of the CNS healthy and happy.  Ken Matoba, Rieko Muramatsu & Toshihide Yamashita. Leptin sustains spontaneous remyelination in the adult central nervous system. Scientific Reports 7, Article number: 40397 (2017) doi:10.1038/srep40397.

Our LepRB Antibody is used in this study to stain OPCs.

Figures: (a) Representative image of cultured OPCs stained with antibodies against LepRb (green) and PDGFRα (red). Scale bar: 25 μm. (b) Relative BrdU incorporation into the OPC obtained from the brain (left graph) and spinal cord (right graph). Cells were treated with recombinant leptin for 48 h (n = 4). (Left graph) P = 0.005993 (control vs 10 ng/mL), 0.045616 (control vs 100 ng/mL), (Right graph) P = 0.004456 (control vs 10 ng/mL), 0.017859 (control vs 100 ng/mL). (c) Relative BrdU incorporation into the OPC after leptin stimulation (10 ng/ml) with U0126 (20 μM), a MEK inhibitor (n = 4 for brain OPCs, n = 3 for spinal cord OPCs). (Left graph) P = 0.019753 (control vs leptin), 0.039433 (leptin vs leptin + U0126), (Right graph) P = 0.045545 (control vs leptin), 0.04486 (leptin vs leptin + U0126). (d) Representative images of western blotting (upper panels) and quantitative analysis of ERK phosphorylation (lower graph) are shown. OPCs were treated with leptin (10 ng/ml) under indicated periods (n = 3). P = 0.006352 (2 min), 0.016571 (5 min), 0.017675 (10 min), 0.024100 (15 min), 0.081342 (30 min).

We are in the process of looking for Labs to sponsor Neuromics' to isolate and purify adult human OPCs in return for receiving 2,000,000 cells. Stay tuned.

Human Astroglia and Schwann Cells-BBB Model

Broadening our Capabilities

At the core of our solutions are many options for primary human cells. We are especially pleased that we have growing capabilities to provide new cells to researchers studying autoimmune neuro-degenerative diseases like ALS and MS with the addition of:
Schwann Cells

Human Schwann Cells (HSwC) are isolated from human spinal nerve. HSwC are cryopreserved at passage one and delivered frozen. Each vial contains >5 x 10^5 cells in 1 ml volume. HSwC are characterized by immunofluorescence with antibodies specific to S100, GFAP, and CD90. HSwC are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. HSwC can to further expand for 10 population doublings in our Schwann Growth medium (cat # SGM001).
Human Astrocytes

 Human Brain Astrocytes cultured with AlphaBioCoat.
Human Blood Brain Barrier Model
I will continue to post updates here.

Apoptosis and Neurodegeneration

Towards the Development of Disease Specific Assays
Neurodegenerative diseases are becoming increasingly prevalent, especially in the Western societies, with larger percentage of members living to an older age. They have to be seen not only as a health problem, but since they are care-intensive, they also carry a significant economic burden.

Apoptotic pathways are induced in many of these diseases and are key culprits in disease progression.

Figure: Schematic representation of apoptotic pathways. Apoptosis triggered by internal (intrinsic) or external (extrinsic) stress signals that is activated by binding of ligands (e.g. FasL, APO-2L, TRAIL, TNF) to cell surface receptors (e.g. Fas, DR4, DR5, TNF-R1). The intrinsic apoptosis pathway might be triggered by p53 upon DNA damage following exposure to cellular stress. In the intrinsic pathway, death signal reaches mitochondria, leading to release of cytochrome c, which can binds to Apaf1. The cytochrome c/Apaf1 make a complex with pro-caspase-9 (in the presence of dATP), activates caspase-9, which promotes caspase-3 activation, eventually leading to cell death. The extrinsic pathway is initiated through the stimulation of the members of tumor necrosis factor receptor (TNF-R) family (transmembrane death receptors) by their respective ligands. These receptors activate pro-caspases-8, -10 by recruiting the endogenous adaptor protein FADD. Procaspase-8, -10 cleave themselves to form activated caspase-8 or -10. Ultimately, effector enzymes such as caspase-3, -6, -7 are activated in this cascade to mediate apoptosis. Likewise, there can be cross-talk between the intrinsic and extrinsic pathways. For example caspase-8 may cleave Bid to form tBid that is a strong activator of the intrinsic/mitochondrial apoptotic pathway. The intrinsic pathway is usually activated by the recruitment of BAX and BAK to outer mitochondrial membrane, causing cytochrome c release formation of apoptosome and subsequent activation of caspase-9. Activated caspase-9 proteolytically activates caspases-3, -6, and -7. Moreover, some of the effector caspases also can activate caspase-8, forming a positive amplification loop. doi:10.1016/j.pneurobio.2013.10.004.

Working with the Apoptosis Experts at Immunochemistry Technologies and Human Astroglial-Neuron Biosensors Experts at ArunA Biomedical, we plan on  developing disease specific assays. Here's a map of the general process.

Larger Image

We welcome feedback and input on your interests. You can email rose@neuromics.com. We will continue to post updates on exciting new developments.

GFP Labeled Motor Neurons and MEA

Big Upcoming Webinar

We have been strategically partnering with ArunA Biomedical to improve how we serve Neuro-drug discovery and Neurotox research community. I am especially excited about how well are our GFP Labeled Mouse Neurons are working in many different and unique research applications. They have proved an important tool in the study of neuro-muscular diseases like ALS, Parkinson's and Multiple Sclerosis.

I am pleased to announce a coming Webinar: “GFP+ Motor Neurons: Development and in-vitro Functional Assessment” Wednesday June 10th, 11:30 AM EDT-Register Today!

Download Flyer. I will continue to post unique assays developed for our Astro-glial Neuron solutions.

Solutions for Studying Neuro-degeneration

Data Rich and Frequently Published

The Neuromics' brand is built, in part, by our proven ability to provide solutions for the study of neuro-degeneration. These include:

• Neuronal-Glial Markers
• Primary Neurons and Astrocytes
• Apoptosis Research Reagents
• Transfection Reagents

A recent example shows the use of one of  MAP-2 markers to study hearing decline with age: Radtke-schuller S, Seeler S and Grothe B(2015) Restricted loss of olivocochlear but not vestibular efferent neurons in the senescent gerbil (Meriones unguiculatus). Front. Aging Neurosci. 7:4. doi:10.3389/fnagi.2015.00004.

Figure: Lipofuscin granules in MSO neurons of an aged gerbil. MSO neurons are MAP2 immunostained (Alexa Fluor 647, red). Lipofuscin granules have been excited with the DAPI excitation wavelengths and appear blue. Confocal images show a maximum projection of image stacks in A and a single optical image of 0.3 µm thickness in the enlargement in B. Scale bar in A: 50 µm and 20 µm in B.

We stand ready to serve you. Should you have interest or questions, do not hesitate to contact me directly: Pete Shuster-Owner/CEO-pshuster@neuromics.com or direct phone: 612-801-1007. Thank you.

SMG-1 and Parkinson's Disease (PD)

Absence of SMG1 protein could lead to PD


A new study has suggested that the absence of a protein called SMG1 - identified as a Regulator of Parkinson's disease-associated alpha-Synuclein-could aid in the development of Parkinson's and other related neurological disorders.

In light of these findings, we believe SMG1 will have increasing importance for PD Researchers. We now have a solid marker for this protein.

Image: Immunoperoxidase of monoclonal antibody to SMG1 on formalin-fixed paraffin-embedded human adrenal gland. [antibody concentration 1.5 ug/ml inset: : Western blot of SMG1 expression in HeLa NE.

Please check out our comprehensive catalog of markers for Parkinson's.

Glutaredoxin 2 prevents aggregation of mutant SOD1

Our PTEN-induced kinase, PINK1 or PARK6 Antibody is an excellent marker for Amyotrophic Lateral Sclerosis (ALS) and Parkinson's Disease (PD) researchers.

Here's  new publication referencing use of this antibody:  Alberto Ferri, Paolo Fiorenzo, Monica Nencini, Mauro Cozzolino, Maria Grazia Pesaresi, Cristiana Valle, Sara Sepe, Sandra Moreno, and Maria Teresa Carrì. Glutaredoxin 2 prevents aggregation of mutant SOD1 in mitochondria and abolishes its toxicity.
Hum. Mol. Genet., first published on Sep 20, 2010 as doi: doi:10.1093/hmg/ddq383

Abstract:
Vulnerability of motoneurons in amyotrophic lateral sclerosis (ALS) arises from a combination of several mechanisms, including protein misfolding and aggregation, mitochondrial dysfunction and oxidative damage. Protein aggregates are found in motoneurons in models for ALS linked to a mutation in the gene coding for Cu,Zn superoxide dismutase (SOD1) and in ALS patients as well. Aggregation of mutant SOD1 in the cytoplasm and/or into mitochondria has been repeatedly proposed as a main culprit for the degeneration of motoneurons. It is, however, still debated whether SOD1 aggregates represent a cause, a correlate or a consequence of processes leading to cell death. We have exploited the ability of glutaredoxins (Grxs) to reduce mixed disulfides to protein thiols either in the cytoplasm and in the IMS (Grx1) or in the mitochondrial matrix (Grx2) as a tool for restoring a correct redox environment and preventing the aggregation of mutant SOD1. Here we show that the overexpression of Grx1 increases the solubility of mutant SOD1 in the cytosol but does not inhibit mitochondrial damage and apoptosis induced by mutant SOD1 in neuronal cells (SH-SY5Y) or in immortalized motoneurons (NSC-34). Conversely, the overexpression of Grx2 increases the solubility of mutant SOD1 in mitochondria, interferes with mitochondrial fragmentation by modifying the expression pattern of proteins involved in mitochondrial dynamics, preserves mitochondrial function and strongly protects neuronal cells from apoptosis. The toxicity of mutant SOD1, therefore, mostly arises from mitochondrial dysfunction and rescue of mitochondrial damage may represent a promising therapeutic strategy.

Related Reagents: Parkin Parkin-2 PARK2 Co-regulated (PACRG) PARK7 (DJ-1) LRRK2 (PARK8)                         Neurodegenerative Disease Research Antibodies

Neurodegenerative Disease Research Proteins Neurotransmission -Neurotransmission Research Antibody Categories                          Neurotrophins and Growth Factor Antibodies Neuron-Glial Expressed-Includes Neurotrophin Proteins

Apoptosis Research Reagents-Apoptosis Categories-includes: detection kits, antibodies and proteins Primary Neurons and Astrocytes -Primary human, rat and mouse neurons and astrocytes by Category

Otx2 (Orthodenticle Homeobox 2) and Parkinson's Disease

Our Otx2 Antibody is a potent marker for Human, Mouse and Rat Midbrain Dopamanergic Progenitors.

This is confirmed by a recent publication by Dr. Ole Isaacson et al:

Chee Yeun Chung, Pawel Licznerski, Kambiz N. Alavian, Antonio Simeone, Zhicheng Lin, Eden Martin, Jeffery Vance and Ole Isacson. The transcription factor orthodenticle homeobox 2 influences axonal projections and vulnerability of midbrain dopaminergic neurons. Brain Advance Access published online on June 23, 2010 Brain, doi:10.1093/brain/awq142... anti-Otx2 (Neuromics, 1:500)...

Abstract: Two adjacent groups of midbrain dopaminergic neurons, A9 (substantia nigra pars compacta) and A10 (ventral tegmental area), have distinct projections and exhibit differential vulnerability in Parkinson’s disease. Little is known about transcription factors that influence midbrain dopaminergic subgroup phenotypes or their potential role in disease. Here, we demonstrate elevated expression of the transcription factor orthodenticle homeobox 2 in A10 dopaminergic neurons of embryonic and adult mouse, primate and human midbrain. Overexpression of orthodenticle homeobox 2 using lentivirus increased levels of known A10 elevated genes, including neuropilin 1, neuropilin 2, slit2 and adenylyl cyclase-activating peptide in both MN9D cells and ventral mesencephalic cultures, whereas knockdown of endogenous orthodenticle homeobox 2 levels via short hairpin RNA reduced expression of these genes in ventral mesencephalic cultures. Lack of orthodenticle homeobox 2 in the ventral mesencephalon of orthodenticle homeobox 2 conditional knockout mice caused a reduction of midbrain dopaminergic neurons and selective loss of A10 dopaminergic projections. Orthodenticle homeobox 2 overexpression protected dopaminergic neurons in ventral mesencephalic cultures from Parkinson’s disease-relevant toxin, 1-methyl-4-phenylpyridinium, whereas downregulation of orthodenticle homeobox 2 using short hairpin RNA increased their susceptibility. These results show that orthodenticle homeobox 2 is important for establishing subgroup phenotypes of post-mitotic midbrain dopaminergic neurons and may alter neuronal vulnerability.

Image: Characterization of the human neuroectodermal precursors. Otx2 Staining of forebrain-midbrain rosettes (dilution 1:1000).

All Publications Referencing Neuromics Purified Goat Polyclonal Otx2

Related Reagents to Consider:

• Otx2-Mouse 
• Stem Cell Research Reagents

NSE and TUJ-1 and Parkinson's Disease Research

I would like to thank Meghan Coakley from University College Cork for alerting me to a new publication referencing our Chicken NSE (Neuron-Specific Enolase) and Tuj 1 (Neuron-specific class III beta-tubulin) antibodies.

Here's her feedback: "Just letting you know we published our paper using the beta-III-Tubulin and NSE antibodies you supplied to us – Timmons et al., Neuroscience Letters, Oct 1, 2009 [Epub ahead of print]. The antibodies were excellent and I’m sure we’ll be using Neuromics again in the future."

Timmons S, Coakley MF, Moloney AM, O' Neill C. Akt signal transduction dysfunction in Parkinson's disease. Neurosci Lett. 2009 Oct 1. [Epub ahead of print].

Abstract: Significant attention has been drawn to the potential role of defective PI3-kinase-Akt (PKB) signalling in Parkinson's disease (PD) neurodegeneration and to the possibility that activation of Akt may provide neuroprotection in PD. However, little knowledge exists on the integrity of the Akt system in PD. Results of the present study show diminished levels of both total and active phospho(Ser473)-Akt in the brain in PD. This was evident by western blot analysis of midbrain fractions from PD compared to non-PD control brain, but more specifically by immunofluorescence microscopy of the substantia nigra pars compacta (SNpc). Here, double immunofluorescence microscopy found Akt and phospho(Ser473)-Akt to be expressed at high levels in tyrosine hydroxylase (TH) immunopositive dopaminergic neurons in control human brain. Selective loss of these neurons was accompanied by a marked decrease of Akt and phospho(Ser473)-Akt expression in the PD brain, however Akt and active phospho(Ser473)-Akt are still evident in degenerating dopaminergic neurons in the disease. This suggests that it may be possible to target neuronal Akt in advanced PD. Converse to the marked loss of neuronal Akt in PD, increased Akt and phospho(Ser473)-Akt levels were observed in small non-TH positive cells in PD SNpc, whose increased number and small nuclear size indicate they are glia. These findings implicate defective Akt as a putative signalling pathway linked to loss of dopaminergic neurons in PD.

Other Reagents to Consider:
Tuj 1 (Neuron-specific class III beta-tubulin)-Mouse Monoclonal
Neuron/Glial Marker Antibodies
Neurotrophins-Neuron/Glial Marker Proteins
Neurodegenerative Disease Research Antibodies
Neurodegenerative Disease Research Proteins
Stem Cell Research Reagents