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Mutual stimulatory signaling between human myogenic cells and rat cerebellar neurons

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Insight into the bidirectional signaling between primary human myogenic cells and neurons is lacking. For this purpose, human myogenic cells were derived from the semitendinosus and gracilis muscles of five healthy individuals and co-cultured with cerebellar granule neurons from two litters of 7-day-old Wistar rat pups, in muscle medium or neural medium, alongside monocultures of myogenic cells or neurons. RT-PCR was performed to determine human mRNA levels of GAPDH, Ki67, myogenin, and MUSK, and the acetylcholine receptor subtypes CHRNA1, CHRNB1, CHRNG, CHRND, and CHRNE, and rat mRNA levels of GAPDH, Fth1, Rack1, vimentin, Cdh13, and Ppp1r1a. Immunocytochemistry was used to evaluate neurite outgrowth (GAP43) in the presence and absence of myogenic cells. Co-culture with primary neurons lead to higher myogenic cell gene expression levels of GAPDH, myogenin, MUSK, CHRNA1, CHRNG, and CHRND, compared to myogenic cells cultured alone. It appeared that neurons preferentially attached to myotubes and that neurite outgrowth was enhanced when neurons were cultured with myogenic cells compared to monoculture. In neural medium, rat mRNA levels of GAPDH, vimentin, Cdh13, and Ppp1r1a were greater in co-culture, versus monoculture, whereas in muscle medium co-culture lead to lower levels of Fth1, Rack1, vimentin, and Cdh13 than monoculture. These findings demonstrate mutually beneficial stimulatory signaling between rat cerebellar granule neurons and human myogenic cells, providing support for an active role for both the neuron and the muscle cell in stimulating neurite growth and myogenesis. Bidirectional muscle nerve signaling.

OriginalsprogEngelsk
Artikelnummere15077
TidsskriftPhysiological Reports
Vol/bind9
Udgave nummer21
Sider (fra-til)e15077
ISSN2051-817X
DOI
StatusUdgivet - nov. 2021

Bibliografisk note

Funding Information:
The authors warmly acknowledge Monika Lucia Bayer and Anja Jokipii‐Utzon for technical assistance with the cell culture work and PCR, respectively. The authors gratefully acknowledge Lundbeck Foundation (R344‐2020‐254), Danish Agency for Culture (FPK.2018‐0036), and the Nordea Foundation (Center for Healthy Aging grant) for providing financial support for the study. The authors acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen, where the slide scanner images were taken.

Funding Information:
The authors warmly acknowledge Monika Lucia Bayer and Anja Jokipii-Utzon for technical assistance with the cell culture work and PCR, respectively. The authors gratefully acknowledge Lundbeck Foundation (R344-2020-254), Danish Agency for Culture (FPK.2018-0036), and the Nordea Foundation (Center for Healthy Aging grant) for providing financial support for the study. The authors acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen, where the slide scanner images were taken.

Publisher Copyright:
© 2021 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society

ID: 75521832