Key Specifications Table
|Species Reactivity||Key Applications||Host||Format||Antibody Type|
|H, R, M, Fe, Ft, Sqd, Dr, Ml||ELISA, IF, IHC, IH(P), IP, WB||Rb||Affinity Purified||Polyclonal Antibody|
|Presentation||Purified rabbit polyclonal in buffer containing liquid in 10 mM HEPES (pH 7.5), 150 mM NaCl (pH 7.5), 100 ug/mL BSA and 50% glycerol.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Maintain frozen at -20ºC to -70ºC in undiluted aliquots for up to 6 months after date of receipt. Avoid repeated freeze/thaw cycles. Do not store in a self defrosting freezer.|
|Material Size||100 µL|
Anti-Tyrosine Hydroxylase Antibody SDS
|Anti-Tyrosine Hydroxylase - 2115918||2115918|
|Anti-Tyrosine Hydroxylase - 2388829||2388829|
|Anti-Tyrosine Hydroxylase - 2392225||2392225|
|Anti-Tyrosine Hydroxylase - 2426518||2426518|
|Anti-Tyrosine Hydroxylase - 2458991||2458991|
|Anti-Tyrosine Hydroxylase - 1951919||1951919|
|Anti-Tyrosine Hydroxylase - 1994651||1994651|
|Anti-Tyrosine Hydroxylase - 2021194||2021194|
|Anti-Tyrosine Hydroxylase - 2038257||2038257|
|Reference overview||Application||Species||Pub Med ID|
|The Progressive BSSG Rat Model of Parkinson's: Recapitulating Multiple Key Features of the Human Disease.|
Van Kampen, JM; Baranowski, DC; Robertson, HA; Shaw, CA; Kay, DG
PloS one 10 e0139694 2015
The development of effective neuroprotective therapies for Parkinson's disease (PD) has been severely hindered by the notable lack of an appropriate animal model for preclinical screening. Indeed, most models currently available are either acute in nature or fail to recapitulate all characteristic features of the disease. Here, we present a novel progressive model of PD, with behavioural and cellular features that closely approximate those observed in patients. Chronic exposure to dietary phytosterol glucosides has been found to be neurotoxic. When fed to rats, β-sitosterol β-d-glucoside (BSSG) triggers the progressive development of parkinsonism, with clinical signs and histopathology beginning to appear following cessation of exposure to the neurotoxic insult and continuing to develop over several months. Here, we characterize the progressive nature of this model, its non-motor features, the anatomical spread of synucleinopathy, and response to levodopa administration. In Sprague Dawley rats, chronic BSSG feeding for 4 months triggered the progressive development of a parkinsonian phenotype and pathological events that evolved slowly over time, with neuronal loss beginning only after toxin exposure was terminated. At approximately 3 months following initiation of BSSG exposure, animals displayed the early emergence of an olfactory deficit, in the absence of significant dopaminergic nigral cell loss or locomotor deficits. Locomotor deficits developed gradually over time, initially appearing as locomotor asymmetry and developing into akinesia/bradykinesia, which was reversed by levodopa treatment. Late-stage cognitive impairment was observed in the form of spatial working memory deficits, as assessed by the radial arm maze. In addition to the progressive loss of TH+ cells in the substantia nigra, the appearance of proteinase K-resistant intracellular α-synuclein aggregates was also observed to develop progressively, appearing first in the olfactory bulb, then the striatum, the substantia nigra and, finally, hippocampal and cortical regions. The slowly progressive nature of this model, together with its construct, face and predictive validity, make it ideal for the screening of potential neuroprotective therapies for the treatment of PD.
|Prenatal exposure to methylphenidate affects the dopamine system and the reactivity to natural reward in adulthood in rats.|
Lepelletier, FX; Tauber, C; Nicolas, C; Solinas, M; Castelnau, P; Belzung, C; Emond, P; Cortese, S; Faraone, SV; Chalon, S; Galineau, L
The international journal of neuropsychopharmacology / official scientific journal of the Collegium Internationale Neuropsychopharmacologicum (CINP) 18 2015
Methylphenidate (MPH) is a commonly-used medication for the treatment of children with Attention-Deficit/Hyperactivity Disorders (ADHD). However, its prescription to adults with ADHD and narcolepsy raises the question of how the brain is impacted by MPH exposure during pregnancy. The goal of this study was to elucidate the long-term neurobiological consequences of prenatal exposure to MPH using a rat model.We focused on the effects of such treatment on the adult dopamine (DA) system and on the reactivity of animals to natural rewards.This study shows that adult male rats prenatally exposed to MPH display elevated expression of presynaptic DA markers in the DA cell bodies and the striatum. Our results also suggest that MPH-treated animals could exhibit increased tonic DA activity in the mesolimbic pathway, altered signal-to-noise ratio after a pharmacological stimulation, and decreased reactivity to the locomotor effects of cocaine. Finally, we demonstrated that MPH rats display a decreased preference and motivation for sucrose.This is the first preclinical study reporting long-lasting neurobiological alterations of DA networks as well as alterations in motivational behaviors for natural rewards after a prenatal exposure to MPH. These results raise concerns about the possible neurobiological consequences of MPH treatment during pregnancy.
|Sexual dimorphism in the hypophysiotropic tyrosine hydroxylase-positive neurons in the preoptic area of the teleost, Clarias batrachus.|
Saha, S; Patil, S; Singh, U; Singh, O; Singru, PS
Biology of sex differences 6 23 2015
Dopamine (DA) neurons in the anteroventral periventricular nucleus (AVPV) in the preoptic area (POA) of mammals express estrogen receptors, regulate luteinizing hormone (LH) secretion, and show distinct sexual dimorphism. In teleosts, hypophysiotropic DA neurons of the nucleus preopticus periventricularis (NPP), located in the anteroventral POA, express estrogen receptors, innervate LH cells, and emerged as a neuroanatomical substrate for inhibiting LH cells. Interestingly, the NPP and AVPV seem to share several similarities. Whether DAergic neurons in the NPP show sexual dimorphism is, however, not known. Based on the proposed homology to AVPV and previous studies showing greater tyrosine hydroxylase (TH) mRNA and enzyme activity levels in the brain of female catfish, we hypothesize that females have greater number of DAergic neurons in the NPP and correspondingly more TH-immunoreactive fiber innervation of the pituitary.Adult, male and female Clarias batrachus collected during the prespawning phase of their reproductive cycle were used. Fish were anesthetized and perfused transcardially with phosphate-buffered saline (pH 7.4) and 4 % paraformaldehyde in phosphate buffer. Sections through the rostro-caudal extent of the POA and pituitary were processed for TH immunofluorescence. Using double immunofluorescence, the association between TH-immunoreactive fibers and LH cells in the pituitary was explored. Sections were analyzed using semiquantitative analysis.NPP in POA of C. batrachus has two distinct subdivisions, viz, anterior (NPPa) and posterior (NPPp), and TH neurons were observed in both the subdivisions. Compared to that in the males, a significantly higher (P less than 0.05) number of TH neurons was consistently observed in the NPPa of females. TH neurons in NPPp, however, showed no difference in the number or immunoreactivity. Since DA neurons in NPPa are hypophysiotropic, we compared TH-fiber innervation of the pituitary in both sexes. Compared to males, proximal pars distalis and LH cells in this region of the pituitary in females were densely innervated by TH fibers.Neurons of NPPa and their innervation to the pituitary seem to be a distinct sexually dimorphic DAergic system in C. batrachus. The DAergic system may serve as a component of the neural mechanisms controlling the sexually dimorphic LH surge in teleosts. Given the similarities shared by NPPa and AVPV, homology between these two nuclei is suggested.
|Catecholamine metabolism in paraganglioma and pheochromocytoma: similar tumors in different sites?|
Grouzmann, E; Tschopp, O; Triponez, F; Matter, M; Bilz, S; Brändle, M; Drechser, T; Sigrist, S; Zulewski, H; Henzen, C; Fischli, S; Abid, K
PloS one 10 e0125426 2015
Pheochromocytoma (PHEO) and paraganglioma (PGL) are catecholamine-producing neuroendocrine tumors that arise respectively inside or outside the adrenal medulla. Several reports have shown that adrenal glucocorticoids (GC) play an important regulatory role on the genes encoding the main enzymes involved in catecholamine (CAT) synthesis i.e. tyrosine hydroxylase (TH), dopamine β-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT). To assess the influence of tumor location on CAT metabolism, 66 tissue samples (53 PHEO, 13 PGL) and 73 plasma samples (50 PHEO, 23 PGL) were studied. Western blot and qPCR were performed for TH, DBH and PNMT expression. We found a significantly lower intra-tumoral concentration of CAT and metanephrines (MNs) in PGL along with a downregulation of TH and PNMT at both mRNA and protein level compared with PHEO. However, when PHEO were partitioned into noradrenergic (NorAd) and mixed tumors based on an intra-tumoral CAT ratio (NE/E greater than 90%), PGL and NorAd PHEO sustained similar TH, DBH and PNMT gene and protein expression. CAT concentration and composition were also similar between NorAd PHEO and PGL, excluding the use of CAT or MNs to discriminate between PGL and PHEO on the basis of biochemical tests. We observed an increase of TH mRNA concentration without correlation with TH protein expression in primary cell culture of PHEO and PGL incubated with dexamethasone during 24 hours; no changes were monitored for PNMT and DBH at both mRNA and protein level in PHEO and PGL. Altogether, these results indicate that long term CAT synthesis is not driven by the close environment where the tumor develops and suggest that GC alone is not sufficient to regulate CAT synthesis pathway in PHEO/PGL.
|Neurite Mistargeting and Inverse Order of Intraretinal Vascular Plexus Formation Precede Subretinal Vascularization in Vldlr Mutant Mice.|
Johnson, V; Xiang, M; Chen, Z; Junge, HJ
PloS one 10 e0132013 2015
In the retina blood vessels are required to support a high metabolic rate, however, uncontrolled vascular growth can lead to impaired vision and blindness. Subretinal vascularization (SRV), one type of pathological vessel growth, occurs in retinal angiomatous proliferation and proliferative macular telangiectasia. In these diseases SRV originates from blood vessels within the retina. We use mice with a targeted disruption in the Vldl-receptor (Vldlr) gene as a model to study SRV with retinal origin. We find that Vldlr mRNA is strongly expressed in the neuroretina, and we observe both vascular and neuronal phenotypes in Vldlr-/- mice. Unexpectedly, horizontal cell (HC) neurites are mistargeted prior to SRV in this model, and the majority of vascular lesions are associated with mistargeted neurites. In Foxn4-/- mice, which lack HCs and display reduced amacrine cell (AC) numbers, we find severe defects in intraretinal capillary development. However, SRV is not suppressed in Foxn4-/-;Vldlr-/- mice, which reveals that mistargeted HC neurites are not required for vascular lesion formation. In the absence of VLDLR, the intraretinal capillary plexuses form in an inverse order compared to normal development, and subsequent to this early defect, vascular proliferation is increased. We conclude that SRV in the Vldlr-/- model is associated with mistargeted neurites and that SRV is preceded by altered retinal vascular development.
|A method for combining RNAscope in situ hybridization with immunohistochemistry in thick free-floating brain sections and primary neuronal cultures.|
Grabinski, TM; Kneynsberg, A; Manfredsson, FP; Kanaan, NM
PloS one 10 e0120120 2015
In situ hybridization (ISH) is an extremely useful tool for localizing gene expression and changes in expression to specific cell populations in tissue samples across numerous research fields. Typically, a research group will put forth significant effort to design, generate, validate and then utilize in situ probes in thin or ultrathin paraffin embedded tissue sections. While combining ISH and IHC is an established technique, the combination of RNAscope ISH, a commercially available ISH assay with single transcript sensitivity, and IHC in thick free-floating tissue sections has not been described. Here, we provide a protocol that combines RNAscope ISH with IHC in thick free-floating tissue sections from the brain and allows simultaneous co-localization of genes and proteins in individual cells. This approach works well with a number of ISH probes (e.g. small proline-rich repeat 1a, βIII-tubulin, tau, and β-actin) and IHC antibody stains (e.g. tyrosine hydroxylase, βIII-tubulin, NeuN, and glial fibrillary acidic protein) in rat brain sections. In addition, we provide examples of combining ISH-IHC dual staining in primary neuron cultures and double-ISH labeling in thick free-floating tissue sections from the brain. Finally, we highlight the ability of RNAscope to detect ectopic DNA in neurons transduced with viral vectors. RNAscope ISH is a commercially available technology that utilizes a branched or "tree" in situ method to obtain ultrasensitive, single transcript detection. Immunohistochemistry is a tried and true method for identifying specific protein in cell populations. The combination of a sensitive and versatile oligonucleotide detection method with an established and versatile protein assay is a significant advancement in studies using free-floating tissue sections.
|Tfap2a and 2b act downstream of Ptf1a to promote amacrine cell differentiation during retinogenesis.|
Jin, K; Jiang, H; Xiao, D; Zou, M; Zhu, J; Xiang, M
Molecular brain 8 28 2015
Retinogenesis is a precisely controlled developmental process during which different types of neurons and glial cells are generated under the influence of intrinsic and extrinsic factors. Three transcription factors, Foxn4, RORβ1 and their downstream effector Ptf1a, have been shown to be indispensable intrinsic regulators for the differentiation of amacrine and horizontal cells. At present, however, it is unclear how Ptf1a specifies these two cell fates from competent retinal precursors. Here, through combined bioinformatic, molecular and genetic approaches in mouse retinas, we identify the Tfap2a and Tfap2b transcription factors as two major downstream effectors of Ptf1a. RNA-seq and immunolabeling analyses show that the expression of Tfap2a and 2b transcripts and proteins is dramatically downregulated in the Ptf1a null mutant retina. Their overexpression is capable of promoting the differentiation of glycinergic and GABAergic amacrine cells at the expense of photoreceptors much as misexpressed Ptf1a is, whereas their simultaneous knockdown has the opposite effect. Given the demonstrated requirement for Tfap2a and 2b in horizontal cell differentiation, our study thus defines a Foxn4/RORβ1-Ptf1a-Tfap2a/2b transcriptional regulatory cascade that underlies the competence, specification and differentiation of amacrine and horizontal cells during retinal development.
|NMDA receptor GluN2A/GluN2B subunit ratio as synaptic trait of levodopa-induced dyskinesias: from experimental models to patients.|
Mellone, M; Stanic, J; Hernandez, LF; Iglesias, E; Zianni, E; Longhi, A; Prigent, A; Picconi, B; Calabresi, P; Hirsch, EC; Obeso, JA; Di Luca, M; Gardoni, F
Frontiers in cellular neuroscience 9 245 2015
Levodopa-induced dyskinesias (LIDs) are major complications in the pharmacological management of Parkinson's disease (PD). Abnormal glutamatergic transmission in the striatum is considered a key factor in the development of LIDs. This work aims at: (i) characterizing N-methyl-D-aspartate (NMDA) receptor GluN2A/GluN2B subunit ratio as a common synaptic trait in rat and primate models of LIDs as well as in dyskinetic PD patients; and (ii) validating the potential therapeutic effect of a cell-permeable peptide (CPP) interfering with GluN2A synaptic localization on the dyskinetic behavior of these experimental models of LIDs. Here we demonstrate an altered ratio of synaptic GluN2A/GluN2B-containing NMDA receptors in the striatum of levodopa-treated dyskinetic rats and monkeys as well as in post-mortem tissue from dyskinetic PD patients. The modulation of synaptic NMDA receptor composition by a cell-permeable peptide interfering with GluN2A subunit interaction with the scaffolding protein postsynaptic density protein 95 (PSD-95) leads to a reduction in the dyskinetic motor behavior in the two animal models of LIDs. Our results indicate that targeting synaptic NMDA receptor subunit composition may represent an intriguing therapeutic approach aimed at ameliorating levodopa motor side effects.
|The temporary and accumulated effects of transcranial direct current stimulation for the treatment of advanced Parkinson's disease monkeys.|
Li, H; Lei, X; Yan, T; Li, H; Huang, B; Li, L; Xu, L; Liu, L; Chen, N; Lü, L; Ma, Y; Xu, L; Li, J; Wang, Z; Zhang, B; Hu, X
Scientific reports 5 12178 2015
Transcranial direct current stimulation (tDCS) is a useful noninvasive technique of cortical brain stimulation for the treatment of neurological disorders. Clinical research has demonstrated tDCS with anodal stimulation of primary motor cortex (M1) in Parkinson's disease (PD) patients significantly improved their motor function. However, few studies have been focused on the optimization of parameters which contributed significantly to the treatment effects of tDCS and exploration of the underline neuronal mechanisms. Here, we used different stimulation parameters of anodal tDCS on M1 for the treatment of aged advanced PD monkeys induced with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) administration, and then analyzed the temporary and accumulated effects of tDCS treatment. The results indicated anodal tDCS on M1 very significantly improved motor ability temporarily; importantly, the treatment effects of anodal tDCS on M1 were quantitatively correlated to the accumulated stimulation instead of the stimuli intensity or duration respectively. In addition, c-fos staining showed tDCS treatment effects activated the neurons both in M1 and substantia nigra (SN). Therefore, we propose that long time and continue anodal tDCS on M1 is a better strategy to improve the motor symptoms of PD than individual manipulation of stimuli intensity or duration.
|Analysing human neural stem cell ontogeny by consecutive isolation of Notch active neural progenitors.|
Edri, R; Yaffe, Y; Ziller, MJ; Mutukula, N; Volkman, R; David, E; Jacob-Hirsch, J; Malcov, H; Levy, C; Rechavi, G; Gat-Viks, I; Meissner, A; Elkabetz, Y
Nature communications 6 6500 2015
Decoding heterogeneity of pluripotent stem cell (PSC)-derived neural progeny is fundamental for revealing the origin of diverse progenitors, for defining their lineages, and for identifying fate determinants driving transition through distinct potencies. Here we have prospectively isolated consecutively appearing PSC-derived primary progenitors based on their Notch activation state. We first isolate early neuroepithelial cells and show their broad Notch-dependent developmental and proliferative potential. Neuroepithelial cells further yield successive Notch-dependent functional primary progenitors, from early and midneurogenic radial glia and their derived basal progenitors, to gliogenic radial glia and adult-like neural progenitors, together recapitulating hallmarks of neural stem cell (NSC) ontogeny. Gene expression profiling reveals dynamic stage-specific transcriptional patterns that may link development of distinct progenitor identities through Notch activation. Our observations provide a platform for characterization and manipulation of distinct progenitor cell types amenable for developing streamlined neural lineage specification paradigms for modelling development in health and disease.
|SNAP i.d. 2.0 System Brochure|
|Millipore Tools for Characterizing Induced Pluripotent Stem Cells|