Sudan black: a fast, easy and non‐toxic method to assess myelin repair in demyelinating diseases

Ineichen, Benjamin Victor; Weinmann, Oliver; Good, Nicolas; Plattner, Patricia S.; Wicki, Carla A.; Linnebank, Michael

doi:10.1111/nan.12373

Cited by 13 publications

(5 citation statements)

References 34 publications

(39 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Samples were stained with DAPI (1:20,000) or tomato lectin conjugated with FITC (1:1,000) and processed as described by Rosato Siri et al., (2013) and Codagnone et al., (2015) with modifications. Myelin staining with Sudan black (0.5% in 70% ethanol for 30 min) was performed following Ineichen et al., (2017). Sections chosen for the rostral region of the body of the CC (rostral CC) corresponded to plates 16 and 17 (from bregma 0.48 to 0.20); those for the caudal region of the body of the CC (caudal CC) and somatosensory cortex (SSC) corresponded to plates 28 to 31 (from bregma −2.56 to −3.30); and those for the medial prefrontal cortex (mPFC) corresponded to plates 7 and 8 (from bregma 4.20 to 3.70) according to the atlas of Paxinos and Watson (1986).…”

Section: Methodsmentioning

confidence: 99%

Neurobiological substrates underlying corpus callosum hypoconnectivity and brain metabolic patterns in the valproic acid rat model of autism spectrum disorder

Uccelli

Codagnone

Traetta

et al. 2021

Journal of Neurochemistry

View full text Add to dashboard Cite

Atypical connectivity between brain regions and altered structure of the corpus callosum (CC) in imaging studies supports the long‐distance hypoconnectivity hypothesis proposed for autism spectrum disorder (ASD). The aim of this study was to unveil the CC ultrastructural and cellular changes employing the valproic acid (VPA) rat model of ASD. Male Wistar rats were exposed to VPA (450 mg/kg i.p.) or saline (control) during gestation (embryonic day 10.5), and maturation, exploration, and social behavior were subsequently tested. Myelin content, ultrastructure, and oligodendroglial lineage were studied in the CC at post‐natal days 15 (infant) and 36 (juvenile). As a functional outcome, brain metabolic activity was determined by positron emission tomography. Concomitantly with behavioral deficits in juvenile VPA rats, the CC showed reduced myelin basic protein, conserved total number of axons, reduced percentage of myelinated axons, and aberrant and less compact arrangements of myelin sheath ultrastructure. Mature oligodendrocytes decreased and oligodendrocyte precursors increased in the absence of astrogliosis or microgliosis. In medial prefrontal and somatosensory cortices of juvenile VPA rats, myelin ultrastructure and oligodendroglial lineage were preserved. VPA animals exhibited global brain hypometabolism and local hypermetabolism in brain regions relevant for ASD. In turn, the CC of infant VPA rats showed reduced myelin content but preserved oligodendroglial lineage. Our findings indicate that CC hypomyelination is established during infancy and prior to oligodendroglial pattern alterations, which suggests that axon–oligodendroglia communication could be compromised in VPA animals. Thus, CC hypomyelination may underlie white matter alterations and contribute to atypical patterns of connectivity and metabolism found in ASD.

show abstract

Section: Methodsmentioning

confidence: 99%

Neurobiological substrates underlying corpus callosum hypoconnectivity and brain metabolic patterns in the valproic acid rat model of autism spectrum disorder

Uccelli

Codagnone

Traetta

et al. 2021

Journal of Neurochemistry

View full text Add to dashboard Cite

show abstract

“…To see whether LPA may be involved in MS, we obtained human MS tissue and examined autotaxin expression around the lesion border. Fresh frozen MS tissue was stained with Sudan Black to demarcate demyelinated regions 20 . The tissue was immunostained for ATX and HLA-DR (a marker for inflammatory microglia and macrophages, 21 ).…”

Section: Resultsmentioning

confidence: 99%

Discovery of a brain penetrant small molecule antagonist targeting LPA1 receptors to reduce neuroinflammation and promote remyelination in multiple sclerosis

Poon,

Lorrain,

Stebbins

et al. 2024

Sci Rep

View full text Add to dashboard Cite

Multiple sclerosis (MS) is a chronic neurological disease characterized by inflammatory demyelination that disrupts neuronal transmission resulting in neurodegeneration progressive disability. While current treatments focus on immunosuppression to limit inflammation and further myelin loss, no approved therapies effectively promote remyelination to mitigate the progressive disability associated with chronic demyelination. Lysophosphatidic acid (LPA) is a pro-inflammatory lipid that is upregulated in MS patient plasma and cerebrospinal fluid (CSF). LPA activates the LPA1 receptor, resulting in elevated CNS cytokine and chemokine levels, infiltration of immune cells, and microglial/astrocyte activation. This results in a neuroinflammatory response leading to demyelination and suppressed remyelination. A medicinal chemistry effort identified PIPE-791, an oral, brain-penetrant, LPA1 antagonist. PIPE-791 was characterized in vitro and in vivo and was found to be a potent, selective LPA1 antagonist with slow receptor off-rate kinetics. In vitro, PIPE-791 induced OPC differentiation and promoted remyelination following a demyelinating insult. PIPE-791 further mitigated the macrophage-mediated inhibition of OPC differentiation and inhibited microglial and fibroblast activation. In vivo, the compound readily crossed the blood–brain barrier and blocked LPA1 in the CNS after oral dosing. Direct dosing of PIPE-791 in vivo increased oligodendrocyte number, and in the mouse experimental autoimmune encephalomyelitis (EAE) model of MS, we observed that PIPE-791 promoted myelination, reduced neuroinflammation, and restored visual evoked potential latencies (VEP). These findings support targeting LPA1 for remyelination and encourage development of PIPE-791 for treating MS patients with advantages not seen with current immunosuppressive disease modifying therapies.

show abstract

“…Commonly used SBB alcohol solvents (e.g. ethanol) are not compatible with the proposed method as the solvent of choice should prevent the loss of lipid particles from the slide and be completely inert with respect to the solubility of lipid material in it [6,8] . After destaining, the slide is air dried at room temperature and digitalized using the office scanner or camera.…”

Section: Methodsmentioning

confidence: 99%

Sudan black lipid blot – a rapid and simple method for quantification of lipids in biological samples

Homolak

Joja

Markovic

et al. 2022

Preprint

View full text Add to dashboard Cite

Bioanalytical techniques for the isolation and quantification of total lipids in biological samples are an integral part of lipidomic workflows and widely used tools for metabolic assessment at the cellular and organismic levels. The most widely used protocol for the isolation, extraction, and quantification of total lipids in biological tissues was originally introduced by Folch et al.. It requires a relatively large amount of tissue and large volumes of lipid extracts for reliable assessment of lipid content using the gravimetric technique. Here, we propose a new method to overcome the aforementioned challenges based on the hypothesis that the partitioning coefficient of the widely used lysochrome diazo dye Sudan Black B between the lipid extract and ethylene glycol can be used to indirectly estimate the absolute concentration of lipids. The proposed method demonstrates great precision and linearity, requires minimal equipment, and enables the analysis of total lipid content in biological specimens available only in limited amounts by reducing the requirements for the input quantity by >300-fold for some tissues (e.g. fecal samples).

show abstract

Sudan black: a fast, easy and non‐toxic method to assess myelin repair in demyelinating diseases

Abstract: Our adapted sudan black staining can be used to rapidly and non-toxically screen for remyelinating therapies in demyelinating diseases.

Cited by 13 publications

References 34 publications

Neurobiological substrates underlying corpus callosum hypoconnectivity and brain metabolic patterns in the valproic acid rat model of autism spectrum disorder

Neurobiological substrates underlying corpus callosum hypoconnectivity and brain metabolic patterns in the valproic acid rat model of autism spectrum disorder

Discovery of a brain penetrant small molecule antagonist targeting LPA1 receptors to reduce neuroinflammation and promote remyelination in multiple sclerosis

Sudan black lipid blot – a rapid and simple method for quantification of lipids in biological samples

Contact Info

Product

Resources

About