ClC-2-like Chloride Current Alterations in a Cell Model of Spinal and Bulbar Muscular Atrophy, a Polyglutamine Disease

Martínez-Rojas, Vladimir A.; Jiménez-Garduño, Aura Matilde; Michelatti, Daniela; Tosatto, Laura; Marchioretto, Marta; Arosio, Daniele; Basso, Manuela; Pennuto, Maria; Musio, Carlo

doi:10.1007/s12031-020-01687-5

Cited by 15 publications

(11 citation statements)

References 75 publications

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“…MNPCs differentiated by mean of the protocol described above were dissociated and 2.5 × 10 5 cells were replated in maturation medium on vitronectin-coated 12-mm plastic coverslips (SPL) and allowed to mature for 30 or 60 days. Patch-clamp recordings were obtained from a total number of 40 hiPSC-derived motor neurons in the whole-cell configuration [ 26 , 27 ]. Pipettes were made on a temperature-controlled PIP6 puller (HEKA) from borosilicate glass capillaries (Harvard Apparatus pipette electrodes had resistance ranging 4–6 MΩ when filled with an intracellular solution containing the following: in (mM) 130 K-gluconate, 10 KCl, 0.1 CaCl 2 , 2.0 MgCl 2 , 1.1 EGTA, 10 HEPES, 2.0 Mg-ATP, and 0.2 Na-GTP, pH 7.3 (280 mOsm).…”

Section: Methodsmentioning

confidence: 99%

A Monolayer System for the Efficient Generation of Motor Neuron Progenitors and Functional Motor Neurons from Human Pluripotent Stem Cells

Cutarelli

Martínez-Rojas

Tata

et al. 2021

Cells

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Methods for the conversion of human induced pluripotent stem cells (hiPSCs) into motor neurons (MNs) have opened to the generation of patient-derived in vitro systems that can be exploited for MN disease modelling. However, the lack of simplified and consistent protocols and the fact that hiPSC-derived MNs are often functionally immature yet limit the opportunity to fully take advantage of this technology, especially in research aimed at revealing the disease phenotypes that are manifested in functionally mature cells. In this study, we present a robust, optimized monolayer procedure to rapidly convert hiPSCs into enriched populations of motor neuron progenitor cells (MNPCs) that can be further amplified to produce a large number of cells to cover many experimental needs. These MNPCs can be efficiently differentiated towards mature MNs exhibiting functional electrical and pharmacological neuronal properties. Finally, we report that MN cultures can be long-term maintained, thus offering the opportunity to study degenerative phenomena associated with pathologies involving MNs and their functional, networked activity. These results indicate that our optimized procedure enables the efficient and robust generation of large quantities of MNPCs and functional MNs, providing a valid tool for MNs disease modelling and for drug discovery applications.

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Section: Methodsmentioning

confidence: 99%

A Monolayer System for the Efficient Generation of Motor Neuron Progenitors and Functional Motor Neurons from Human Pluripotent Stem Cells

Cutarelli

Martínez-Rojas

Tata

et al. 2021

Cells

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“…Possibly, a combined treatment of IGF-1 together with an anti-androgen, such as leuprorelin (Banno et al, 2009), may enhance the potential of both approaches to either arrest or at least delay disease progression. Consistent with their neuroprotective effects, both IGF-1 and PACAP ameliorated ionic current defects in a cell model of SBMA (Jimenez Garduno et al, 2017;Martinez-Rojas et al, 2021). The promising therapeutic effects of these peptides may ultimately contribute to therapy development for MNDs.…”

Section: Role Of Igf-1 In Mndsmentioning

confidence: 66%

Motor Neuron Diseases and Neuroprotective Peptides: A Closer Look to Neurons

Zuccaro

Piol

Basso

et al. 2021

Front. Aging Neurosci.

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Motor neurons (MNs) are specialized neurons responsible for muscle contraction that specifically degenerate in motor neuron diseases (MNDs), such as amyotrophic lateral sclerosis (ALS), spinal and bulbar muscular atrophy (SBMA), and spinal muscular atrophy (SMA). Distinct classes of MNs degenerate at different rates in disease, with a particular class named fast-fatigable MNs (FF-MNs) degenerating first. The etiology behind the selective vulnerability of FF-MNs is still largely under investigation. Among the different strategies to target MNs, the administration of protective neuropeptides is one of the potential therapeutic interventions. Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide with beneficial effects in many neurodegenerative diseases, including Alzheimer’s disease, Parkinson’s disease, and more recently SBMA. Another neuropeptide that has a neurotrophic effect on MNs is insulin-like growth factor 1 (IGF-1), also known as somatomedin C. These two peptides are implicated in the activation of neuroprotective pathways exploitable in the amelioration of pathological outcomes related to MNDs.

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“…As PACAP is a cAMP stimulating peptide, it can be assumed that the neuropeptide plays a role endogenously in the aqueous humour production. Furthermore, PACAP and its receptors have also been shown to act on chloride channels, which are essential in the production of aqueous humour, independently from the cAMP pathway (Alsha e et al 2014; Derand et al 2004;Leung et al 2001;Martinez-Rojas et al 2021). The role of PACAP has been implied not only in the production, but also in the absorption of the aqueous humour, as our most recent data have provided evidence that PACAP treatment leads to reduced intraocular pressure in a rat model of glaucoma (Szabo et al 2021).…”

Section: Discussionmentioning

confidence: 99%

Distribution of PACAP and PAC1 Receptor in the Human Eye

Patko

Szabó

Tóth

et al. 2022

Preprint

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Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide with widespread distribution and diverse biological functions. Several studies show that PACAP has strong cytoprotective effects mediated mostly through its specific PAC1 receptor (PAC1-R) and it plays important roles in several pathological conditions. Its distribution and altered expression are known in various human tissues, but there is no descriptive data about PACAP and its receptors in the human eyebulb. Since the PACAP38 is the dominant form of the naturally occurring PACAP, our aim was to investigate the distribution of PACAP38-like immunoreactivity in the human eye and to describe the presence of PAC1-R. Semiquantitative evaluation was performed after routine histology and immunohistochemical labeling on human eye sections. Our results showed high level of immunopositivity in the corneal epithelium and endothelium. Within the vascular layer the iris and the ciliary body had strong immunopositivity for both PACAP and the PAC1-R. Several layers of the retina showed immunoreactivity for PACAP and PAC1-R, but the ganglion cell layer had a special pattern in the immunolabeling. Labeling was observed in the neuropil within the optic nerve in both cases and glial cells displayed immunoreactivity for PAC1-R. In summary, our study indicates the widespread occurrence of PACAP and its specific receptor in the human eye, implying that the results from in vitro and animal studies have translational value and most probably are also present in the human eye.

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ClC-2-like Chloride Current Alterations in a Cell Model of Spinal and Bulbar Muscular Atrophy, a Polyglutamine Disease

Cited by 15 publications

References 75 publications

A Monolayer System for the Efficient Generation of Motor Neuron Progenitors and Functional Motor Neurons from Human Pluripotent Stem Cells

A Monolayer System for the Efficient Generation of Motor Neuron Progenitors and Functional Motor Neurons from Human Pluripotent Stem Cells

Motor Neuron Diseases and Neuroprotective Peptides: A Closer Look to Neurons

Distribution of PACAP and PAC1 Receptor in the Human Eye

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