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Cell transplantation: a future therapy for narcolepsy?
Arias-Carrion O, Murillo-Rodriguez E (2009) Cell transplantation: a future therapy for narcolepsy?. CNS Neurol Disord Drug Targets 8:309-314. doi: 10.2174/187152709788921681
Summary: This review covers the current understanding of narcolepsy and discusses the potential for transplants as a therapeutic treatment. Animal models are summarized, including the use of orexin-SAP (Cat. #IT-20) in rats. The review goes on to suggest that production of orexigenic neuroblasts from stem cells may be a useful therapy.
Related Products: Orexin-B-SAP (Cat. #IT-20)
Basal cells as stem cells of the mouse trachea and human airway epithelium
Rock JR, Onaitis MW, Rawlins EL, Lu Y, Clark CP, Xue Y, Randell SH, Hogan BL (2009) Basal cells as stem cells of the mouse trachea and human airway epithelium. Proc Natl Acad Sci U S A 106(31):12771-12775. doi: 10.1073/pnas.0906850106 PMID: 19625615
Related Products: NGFr (mu p75) Rabbit Polyclonal (Cat. #AB-N01)
Central chemoreception is a complex system function that involves multiple brain stem sites.
Nattie E, Li A (2009) Central chemoreception is a complex system function that involves multiple brain stem sites. J Appl Physiol 106:1464-1466. doi: 10.1152/japplphysiol.00112.2008
Summary: This short review discusses central chemoreception and the different neuronal subtypes that play roles in this process. The use of anti-SERT-SAP (Cat. #IT-23) and anti-DBH-SAP (Cat. #IT-03) is mentioned in the context of how the loss of each of these cell types affects CO2 response in rats.
Related Products: Anti-SERT-SAP (Cat. #IT-23), Anti-DBH-SAP (Cat. #IT-03)
Histone H2AX-dependent GABAA receptor regulation of stem cell proliferation
Andäng M, Hjerling-Leffler J, Moliner A, Lundgren TK, Castelo-Branco G, Nanou E, Pozas E, Bryja V, Halliez S, Nishimaru H, Wilbertz J, Arenas E, Koltzenburg M, Charnay P, Manira AE, Ibañez CF, Ernfors P (2008) Histone H2AX-dependent GABAA receptor regulation of stem cell proliferation. Nature 451(7177):460-464. doi: 10.1038/nature06488 PMID: 18185516
Summary: Immunofluorescence staining for vesicular GABA transporter (VGAT) in embryonic stem cells reveals a punctate localization.
Usage: Immunostaining and western blotting. After being blocked for 2 h in PBS containing 3% BSA and 0.3% Triton X-100 and after two washes in PBS, cells were incubated in primary antibody against VGAT (diluted 1:400).
Related Products: vGAT Rabbit Polyclonal (Cat. #AB-N44)
Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells
Lee G, Kim H, Elkabetz Y, Al Shamy G, Panagiotakos G, Barberi T, Tabar V, Studer L (2007) Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells. Nat Biotechnol 25(12):1468-1475. doi: 10.1038/nbt1365 PMID: 18037878
Usage: Flow Cytometry cell sorting was performed using Anti-NGFr (p75).
Related Products: NGFr (mu p75) Rabbit Polyclonal (Cat. #AB-N01)
Survival, integration and differentiation of human neural stem cells transplanted into an animal model of cholinergic degeneration
Cusulin C, Aztiria E, Cacci E, Battaglini PP, Kokaia Z, Leanza G (2007) Survival, integration and differentiation of human neural stem cells transplanted into an animal model of cholinergic degeneration. Neuroscience 2007 Abstracts 779.5/E7. Society for Neuroscience, San Diego, CA.
Summary: The recently characterized F7B cell line, derived from human fetal cortex (Cacci et al, 2007) has been observed able to differentiate in vitro into different glial and neuronal subtypes. However, no much data is available about its capacity for differentiation in vivo. In the present work, F7B cells were grafted to the medial septum of newborn (P8) intact rats and to littermates that had been subjected to selective cholinergic deafferentation at P4 using the 192 IgG-saporin immunotoxin. The animals were sacrificed 1 or 3 months after grafting and the dissected brains were processed for immunocytochemistry, using cell-specific (HuNu and GFP) and differentiation markers (Dcx, GFAP, NeuN, and HuD). Overall, grafted F7B cells exhibited an excellent ability to survive and differentiate into the host tissue environment. Survival rate varied among the groups, being consistently higher when the cells were grafted into lesioned, as opposed to intact, animal. Moreover, a better survival was seen at 1 month, compared to 3 months post-grafting, regardless the lesion condition. Interestingly, the presence of a cholinergic depletion in the recipient appeared to affect differentiation of grafted F7B cells. In fact, higher numbers of Dcx+ and HuD+ cells were scattered within the grafts placed in lesioned animals, as compared to controls. In lesioned, but not intact animals, at 3 months post-grafting, sparse F7B cells were found to express the mature neuronal marker NeuN. On the other hand, grafted F7B immunoreactive for GFAP were similarly detected in all transplanted animals. F7B cells appear to be feasible for transplantation, being able to survive and differentiate into a developing brain, and to positively respond to the new environment created by a lesion. Further studies are warranted to test their actual capacity for functional integration.
Related Products: 192-IgG-SAP (Cat. #IT-01)
Characterization of an immunotoxin model of Parkinson’s disease in mice
Stead S, Trottier N, Doering LC (2005) Characterization of an immunotoxin model of Parkinson’s disease in mice. Neuroscience 2005 Abstracts 664.9. Society for Neuroscience, Washington, DC.
Summary: The primary event underlying the motor deficits of Parkinson’s disease (PD) is degeneration of neurons in the nigro-striatal system. The most widely employed laboratory rodent models of Parkinson’s are the neurotoxin 6-hydroxydopamine (6-OHDA) model that causes acute degeneration of the dopamine neurons in the substantia nigra (SN) and the MPTP mouse model. To date, there is no single model which accurately simulates the pathogenic, histological, biochemical and clinical features relevant for the investigation of PD. Toxins conveyed by axonal transport can be used to make selective lesions in the central nervous system. As previously shown in rats (Wiley et al., Cell. Mol. Biol., 2003), we have found that selective degeneration of the SN can be induced with an immunotoxin consisting of the highly active ribosome inactivating protein Saporin linked to an antibody to the dopamine transporter. A unilateral stereotaxic injection of anti-DAT-Saporin (0.25ug/2ul and 0.05ug/2ul) into the striatum of young (6-8 weeks old) female C57BL6 mice causes a progressive reduction in the number of DA neurons in the SN in comparison to the non-lesioned hemisphere and in various controls. Furthermore, in parallel to the immunohistochemical dopamine neuron death, the animals display a pronounced circling behaviour when challenged with apomorphine (3mg/kg). We are currently examining the affected brain sections for inclusion bodies and changes in astrocytes. This model exhibits the selective deterioration of the nigro-striatal system that occurs in Parkinson’s disease and provides a system to intervene at various stages of dopamine neuron loss and evaluate the effectiveness of stem cell therapy.
Related Products: Anti-DAT-SAP (Cat. #IT-25)
A novel mouse model for Parkinson’s disease using an immunotoxin directed at the dopamine transporter
Stead S, Doering LC (2004) A novel mouse model for Parkinson’s disease using an immunotoxin directed at the dopamine transporter. Neuroscience 2004 Abstracts 563.1. Society for Neuroscience, San Diego, CA.
Summary: Current laboratory models of Parkinson’s disease utilize neurotoxins directed at midbrain dopamine neurons to mimic nigro-striatal dopaminergic neuron degeneration. To date, however, there is no single model that accurately simulates the pathogenic, histological, biochemical and clinical features relevant for the investigation of PD. The most common laboratory rodent model of Parkinson’s uses the neurotoxin 6-hydroxydopamine (6-OHDA) to cause relatively acute degeneration of the dopamine neurons in the substantia nigra (Schwarting RKW and Huston JP, 1996, Prog Neurobiol., 50:275-331). Axonally transported toxins can be used to make selective lesions in the central nervous system. We have found that a slower degeneration of the SN can be achieved with an immunotoxin directed against the dopamine transporter (DAT). This immunotoxin, consisting of the highly active ribosome inactivating protein Saporin linked to an antibody to the dopamine transporter, was recently reported to cause selective degeneration of the SN in rats (Wiley RG et al., 2003, Cell Mol Neurobiol., 23:839-850.). We have shown that unilateral stereotaxic injection of the Anti-DAT-Saporin into the striatum of female C57BL6 mice causes a progressive reduction in the numbers of DA neurons in the SN in comparison to the non-lesioned hemisphere, and sham controls. Furthermore, in parallel to the immunohistochemical dopamine neuron death, the animals display a pronounced circling behaviour when challenged with apomorphine (6mg/kg). This model is akin to the gradual deterioration of the nigro-striatal system that occurs in Parkinson’s Disease and provides a system to intervene at various stages of dopamine neuron loss and evaluate the effectiveness of stem cell therapy.
Related Products: Anti-DAT-SAP (Cat. #IT-25)
Organized development from human embryonic stem cells after injection into immunodeficient mice
Gertow K, Wolbank S, Rozell B, Sugars R, Andang M, Parish CL, Imreh MP, Wendel M, Ahrlund-Richter L (2004) Organized development from human embryonic stem cells after injection into immunodeficient mice. Stem Cells Dev 13:421-435. doi: 10.1089/scd.2004.13.421 PMID: 15345136
Summary: After thawing and brief drying, sections were fixed with 4% paraformaldehyde in PBS pH 7.4 for 40 min. Sections were blocked [PBS, 1% bovine serum albumin (BSA), 0.3% Triton X-100], washed, and incubated with primary antibodies for Tuj1 (-tubulin type III), glial fibrillary acid protein (GFAP), tyrosine hydroxylase (TH), and vesicular GABA transporter (vGAT; 1:400) overnight at 4°C. After rinsing in PBS, slides were incubated with Cy2- or Cy3-conjugated secondary antibodies.
Related Products: vGAT Rabbit Polyclonal (Cat. #AB-N44)
Eliminating CBF Neurons
Q: What dosage of 192-Saporin (192-IgG-SAP, Cat. #IT-01) should be used in the lateral ventricle to eliminate cholinergic neurons in the basal forebrain, including substantia innominata (SI)? I read that Calza et al [1] used 2 or 3 micrograms/4.5 µl and found this was highly effective.
A: It has been our experience that two- or three-micrograms into the lateral ventricle is necessary to obtain a maximum cholinergic basal forebrain (CBF) lesion. However, these doses typically kill some cerebellar Purkinje cells. Another issue is that some cholinergic neurons in the NBM region are never killed by 192-Saporin.
Q: Should we expect to be able to kill all or almost all ChAT SI neurons?
A: Mesulam’s lab has some data [2,3] to suggest that these neurons innervate the amygdala and adjacent cortex. Generally lesions of the septum and diagonal band are complete, but when you get more caudal, i.e. SI region, there will be some cholinergic neurons left. When you do ChAT or AChE stains, the amygdala and adjacent cortex are not denervated whereas the hippocampus, olfactory system and all the rest of the cortex are devoid of cholinergic terminals.
Q: Is there another toxin that will eliminate the remaining ChAT SI neurons?
A: There may be other targeted conjugates that could clean out the residual cells in the SI region if we knew what markers they co-express. For example, our SSP-saporin (Cat. #IT-11) conjugate is very good at removing cells that express the NK-1 receptor such as striatal cholinergic interneurons.
See: 192-IgG-SAP (Cat. #IT-01), SSP-SAP (Cat. #IT-11), Targeted Toxins
References
- Calza L et al. Neural stem cells and cholinergic neurons: Regulation by immunolesion and treatment with mitogens, retinoic acid, and nerve growth factor. Proc Natl Acad Sci U S A 100(12):7325-7330, 2003.
- Heckers S et al. Two types of cholinergic projections to the rat amygdala. Neuroscience 60:383-397, 1994.
- Heckers S et al. Complete and selective cholinergic denervation of rat neocortex and hippocampus but not amygdala by an immunotoxin against the p75 NGF receptor. J Neurosci 14:1271-1289, 1994.