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Recombinant human GDNF protein (Qk051)
Glial cell line-derived neurotrophic factor (GDNF) is a member of neurotrophin family and GDNF family of ligands (GFL). GDNF plays a crucial role in the development, growth, and survival of neurons in particular midbrain dopaminergic neurons. It promotes the axon growth and innervation of dopamine neurons. GNDF is used to maintain neurons and cortical organoids and to differentiate dopaminergic neurons from human pluripotent stem cell-derived neural progenitors. GDNF also facilitates the differentiation of neural progenitors to astrocytes.
Recombinant human GDNF bioactive 30 kDa homodimer. This protein is animal-free (AOF), carrier protein-free, and tag-free to ensure its purity with exceptional lot-to-lot consistency. Qk051 is suitable for the culture of reproducible and high-quality cortical and motor neurons and cortical organoids.
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1mg will be despatched as 2 x 500µg
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Summary
High purity GDNF (Uniprot: P39905)
>98%, by SDS-PAGE quantitative densitometry
15.1 kDa monomer, 30.4 kDa (dimer)
Expressed in E. coli
Animal-free (AOF) and carrier protein-free
Manufactured in Cambridge, UK
Lyophilized from acetonitrile, TFA
Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20 oC or -80 oC
Featured applications
Differentiation of midbrain dopaminergic neurons
Astrocyte-derived trophic factor, ATF, ATF1, ATF2; glial cell line derived neurotrophic factor; glial derived neurotrophic factor; HFB1-GDNF; HSCR3
Human
Bioactivity
GDNF activity is determined using a SH-SY5Y cell proliferation assay. Cells were incubated with different concentrations of GDNF in the presence of retinoic acid and recombinant GFR α1 for 3 days before viable cell measurement using an MTS assay. Data are n=2.
EC50 = 18 ng/ml
Data from Qk051 batch #104372.
Purity
GDNF migrates as a single band at 30 kDa in non-reducing (NR) conditions and 15 kDa upon reduction (R). No contaminating protein bands are visible.
Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk051 batch #104372.
Further quality assays
Mass spectrometry: single species with expected mass
Endotoxin: <0.005 EU/μg protein (below level of detection)
Recovery from stock vial: >95%
We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine. All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.
Comparison with other suppliers
Qkine GDNF (Qk051) bioactivity was compared with an alternative supplier (Supplier 1) using a SH-SY5Y cell proliferation assay (as above). Qkine GDNF was found to have higher bioactivity than Supplier 1 GDNF. Qk051 EC50 = 18 ng/ml, Supplier 1 EC50 = 26 ng/ml, P = 0.018
Protein background
Glial cell line-derived neurotrophic factor (GDNF) is a neurotrophic factor and a member of the GDNF family of ligands (GFL) which include neurturin, artemin, and persephin1,2. GFLs regulate several biological processes including cell survival, cell proliferation, cell differentiation, cell migration, and neurite outgrowth1,3,4. GDNF protein plays a crucial role in the development, growth, and survival of neurons in particular midbrain dopaminergic neurons5. It promotes the axon growth and innervation of dopamine neurons. It also facilitates the differentiation of neural progenitors to astrocytes and is involved in neuroprotection6,7. In addition, it is involved in kidney development and spermatogenesis8,9.
In cell culture, GDNF supports the survival and growth of neurons and astrocytes. It is used in combination with other growth factors to differentiate human induced pluripotent stem cell-derived neural progenitors into neurons and astrocytes. GDNF is used for the differentiation, survival, and maturation of dopaminergic neurons with brain-derived neurotrophic factor (BDNF), FGF-8a/b, and Shh10–13. Glutamatergic and GABAergic neurons are obtained with combinations of BDNF, GDNF, NT-3, or insulin-like growth factor 1 (IGF-1)14–16. Also, the culture of cortical organoids requires BDNF and GDNF for maturation17,18. Finally, it can be used to generate and maintain astrocytes along with ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF)19–21.
GDNF is mainly produced and released by glial cells such as astrocytes, Schwann cells, and satellite cells22. It is a homodimer of a molecular weight of 30 kDa which belongs to the cystine-knot protein family1,23. The GFLs act as biologically active homodimers that signal through the transmembrane RET receptor tyrosine kinase24. GFL activation of RET is dependent upon co-receptors, namely the four GDNF Family Receptor α (GFRα1-4). GFL signalling specificity arises from the preferentially binding of each ligand to one of the four GFRα receptors; GDNF preferentially binds GFRα1 with high affinity1,24.
GDNF has shown promise in various therapeutic applications neurodegenerative diseases and disorders such as Parkinson’s disease, amyotrophic lateral sclerosis, Huntington’s disease, peripheral nerve and spinal cord injuries1,25. The idea is to harness GDNF for clinical use to promote the survival and function of dopaminergic neurons, potentially slowing or reversing the progression of these diseases. Clinical trials using it has shown improved motor function in patients with Parkinson’s disease26,27. Finally, GDNF has also been proposed for the treatment of drug addiction and alcoholism25.
- Airaksinen, M. S. & Saarma, M. The GDNF family: Signalling, biological functions and therapeutic value. Nat. Rev. Neurosci. 3, 383–394 (2002).
- von dem Bussche, M. & Tuszynski, M. H. Growth Factors: Neuronal Atrophy. in Encyclopedia of Neuroscience (ed. Squire, L. R.) 987–992 (Academic Press, 2009). doi:10.1016/B978-008045046-9.00143-1.
- Airaksinen, M. S., Titievsky, A. & Saarma, M. GDNF Family Neurotrophic Factor Signaling: Four Masters, One Servant? Mol. Cell. Neurosci. 13, 313–325 (1999).
- Baloh, R. H., Enomoto, H., Johnson, E. M. & Milbrandt, J. The GDNF family ligands and receptors — implications for neural development. Curr. Opin. Neurobiol. 10, 103–110 (2000).
- Akerud, P., Alberch, J., Eketjäll, S., Wagner, J. & Arenas, E. Differential effects of glial cell line-derived neurotrophic factor and neurturin on developing and adult substantia nigra dopaminergic neurons. J. Neurochem. 73, 70–78 (1999).
- Parsadanian, A., Pan, Y., Li, W., Myckatyn, T. M. & Brakefield, D. Astrocyte-derived transgene GDNF promotes complete and long-term survival of adult facial motoneurons following avulsion and differentially regulates the expression of transcription factors of AP-1 and ATF/CREB families. Exp. Neurol. 200, 26–37 (2006).
- Boku, S. et al. GDNF facilitates differentiation of the adult dentate gyrus-derived neural precursor cells into astrocytes via STAT3. Biochem. Biophys. Res. Commun. 434, 779–784 (2013).
- Costantini, F. & Shakya, R. GDNF/Ret signaling and the development of the kidney. BioEssays News Rev. Mol. Cell. Dev. Biol. 28, 117–127 (2006).
- Hofmann, M.-C. Gdnf signaling pathways within the mammalian spermatogonial stem cell niche. Mol. Cell. Endocrinol. 288, 95–103 (2008).
- Granholm, A.-C. et al. Glial Cell Line-Derived Neurotrophic Factor Is Essential for Postnatal Survival of Midbrain Dopamine Neurons. J. Neurosci. 20, 3182–3190 (2000).
- Perrier, A. L. et al. Derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl. Acad. Sci. U. S. A. 101, 12543–12548 (2004).
- Brennand, K. et al. Modeling schizophrenia using hiPSC neurons. Nature 473, 221–225 (2011).
- Kriks, S. et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature 480, 547–551 (2011).
- Fernandopulle, M. S. et al. Transcription-factor mediated differentiation of human iPSCs into neurons. Curr. Protoc. Cell Biol. 79, e51 (2018).
- Li, X.-J. et al. Specification of motoneurons from human embryonic stem cells. Nat. Biotechnol. 23, 215–221 (2005).
- Lin, L., Yuan, J., Sander, B. & Golas, M. M. In Vitro Differentiation of Human Neural Progenitor Cells Into Striatal GABAergic Neurons. Stem Cells Transl. Med. 4, 775–788 (2015).
- McComish, S. F. & Caldwell, M. A. Generation of defined neural populations from pluripotent stem cells. Philos. Trans. R. Soc. B Biol. Sci. 373, 20170214 (2018).
- Jacob, F. et al. Human Pluripotent Stem Cell-Derived Neural Cells and Brain Organoids Reveal SARS-CoV-2 Neurotropism Predominates in Choroid Plexus Epithelium. Cell Stem Cell 27, 937-950.e9 (2020).
- Krencik, R., Weick, J. P., Liu, Y., Zhang, Z. & Zhang, S.-C. Specification of Transplantable Astroglial Subtypes from Human Pluripotent Stem Cells. Nat. Biotechnol. 29, 528–534 (2011).
- Serio, A. et al. Astrocyte pathology and the absence of non-cell autonomy in an induced pluripotent stem cell model of TDP-43 proteinopathy. Proc. Natl. Acad. Sci. U. S. A. 110, 4697–4702 (2013).
- Gunhanlar, N. et al. A simplified protocol for differentiation of electrophysiologically mature neuronal networks from human induced pluripotent stem cells. Mol. Psychiatry 23, 1336–1344 (2018).
- Rocha, S. M., Cristovão, A. C., Campos, F. L., Fonseca, C. P. & Baltazar, G. Astrocyte-derived GDNF is a potent inhibitor of microglial activation. Neurobiol. Dis. 47, 407–415 (2012).
- Ibáñez, C. F. Emerging themes in structural biology of neurotrophic factors. Trends Neurosci. 21, 438–444 (1998).
- Takahashi, M. The GDNF/RET signaling pathway and human diseases. Cytokine Growth Factor Rev. 12, 361–373 (2001).
- Hsieh, J. C. & Penn, R. D. Chapter 44 – Infusion Therapy for Movement Disorders. in Neuromodulation (eds. Krames, E. S., Peckham, P. H. & Rezai, A. R.) 561–570 (Academic Press, 2009). doi:10.1016/B978-0-12-374248-3.00045-8.
- Manfredsson, F. P. et al. The Future of GDNF in Parkinson’s Disease. Front. Aging Neurosci. 12, (2020).
- Barker, R. A. et al. GDNF and Parkinson’s Disease: Where Next? A Summary from a Recent Workshop. J. Park. Dis. 10, 875–891.
FAQ
What is the function of GDNF?
This protein plays a crucial role in the development, growth, and survival of midbrain dopaminergic neurons.
Which cells release GDNF?
It is mainly released by glial cells such as astrocytes, Schwann cells, and satellite cells.
What are examples of neurotrophic factors?
Neurotrophic factors (also known as neurotrophins) include Nerve Growth Factor (NGF), Brain-Derived Neurotrophic Factor (BDNF), Glial-Derived Neurotrophic Factor (GDNF), Neurotrophin-3 (NT-3), and Neurotrophin-4/5 (NT-4/5).
How is GDNF used to treat Parkinson’s disease?
GDNF has shown potential therapeutic applications for Parkinson’s disease. Clinical trials using it has shown improved motor function which could slow the progression of the disease.
What is GDNF in cell culture?
It is used as a growth factor which is supplemented in cell culture media to promote the differentiation, growth, and maturation of dopaminergic neurons.
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
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What others are saying
Great product
hh495 –
Very happy with this product. My neuronal cultures look very healthy and there has been no difference in morphology or mechanism of the neurons since switching from a more expensive competitive brand. Our lab will be continuing to purchase.
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