Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons.
A key objective of stem cell biology is to create physiologically relevant cells suitable for modeling disease pathologies in vitro. Much progress towards this goal has been made in the area of motor neuron (MN) disease through the development of methods to direct spinal MN formation from both embry...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3344836?pdf=render |
| _version_ | 1818512854662250496 |
|---|---|
| author | Joy A Umbach Katrina L Adams Cameron B Gundersen Bennett G Novitch |
| author_facet | Joy A Umbach Katrina L Adams Cameron B Gundersen Bennett G Novitch |
| author_sort | Joy A Umbach |
| collection | DOAJ |
| description | A key objective of stem cell biology is to create physiologically relevant cells suitable for modeling disease pathologies in vitro. Much progress towards this goal has been made in the area of motor neuron (MN) disease through the development of methods to direct spinal MN formation from both embryonic and induced pluripotent stem cells. Previous studies have characterized these neurons with respect to their molecular and intrinsic functional properties. However, the synaptic activity of stem cell-derived MNs remains less well defined. In this study, we report the development of low-density co-culture conditions that encourage the formation of active neuromuscular synapses between stem cell-derived MNs and muscle cells in vitro. Fluorescence microscopy reveals the expression of numerous synaptic proteins at these contacts, while dual patch clamp recording detects both spontaneous and multi-quantal evoked synaptic responses similar to those observed in vivo. Together, these findings demonstrate that stem cell-derived MNs innervate muscle cells in a functionally relevant manner. This dual recording approach further offers a sensitive and quantitative assay platform to probe disorders of synaptic dysfunction associated with MN disease. |
| first_indexed | 2024-12-10T23:52:51Z |
| format | Article |
| id | doaj.art-25bf8d71cc134ced9b74f8fd1e4d1c17 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T23:52:51Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-25bf8d71cc134ced9b74f8fd1e4d1c172022-12-22T01:28:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0175e3604910.1371/journal.pone.0036049Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons.Joy A UmbachKatrina L AdamsCameron B GundersenBennett G NovitchA key objective of stem cell biology is to create physiologically relevant cells suitable for modeling disease pathologies in vitro. Much progress towards this goal has been made in the area of motor neuron (MN) disease through the development of methods to direct spinal MN formation from both embryonic and induced pluripotent stem cells. Previous studies have characterized these neurons with respect to their molecular and intrinsic functional properties. However, the synaptic activity of stem cell-derived MNs remains less well defined. In this study, we report the development of low-density co-culture conditions that encourage the formation of active neuromuscular synapses between stem cell-derived MNs and muscle cells in vitro. Fluorescence microscopy reveals the expression of numerous synaptic proteins at these contacts, while dual patch clamp recording detects both spontaneous and multi-quantal evoked synaptic responses similar to those observed in vivo. Together, these findings demonstrate that stem cell-derived MNs innervate muscle cells in a functionally relevant manner. This dual recording approach further offers a sensitive and quantitative assay platform to probe disorders of synaptic dysfunction associated with MN disease.http://europepmc.org/articles/PMC3344836?pdf=render |
| spellingShingle | Joy A Umbach Katrina L Adams Cameron B Gundersen Bennett G Novitch Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons. PLoS ONE |
| title | Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons. |
| title_full | Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons. |
| title_fullStr | Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons. |
| title_full_unstemmed | Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons. |
| title_short | Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons. |
| title_sort | functional neuromuscular junctions formed by embryonic stem cell derived motor neurons |
| url | http://europepmc.org/articles/PMC3344836?pdf=render |
| work_keys_str_mv | AT joyaumbach functionalneuromuscularjunctionsformedbyembryonicstemcellderivedmotorneurons AT katrinaladams functionalneuromuscularjunctionsformedbyembryonicstemcellderivedmotorneurons AT cameronbgundersen functionalneuromuscularjunctionsformedbyembryonicstemcellderivedmotorneurons AT bennettgnovitch functionalneuromuscularjunctionsformedbyembryonicstemcellderivedmotorneurons |