Link to the University of Pittsburgh Homepage
Link to the University Library System Homepage Link to the Contact Us Form

LGMD-1C: Role of Caveolin-3 in Neuromuscular Junction Structure and Function

Hezel, Michael P. (2009) LGMD-1C: Role of Caveolin-3 in Neuromuscular Junction Structure and Function. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

[img]
Preview
PDF
Primary Text

Download (4MB) | Preview

Abstract

Caveolin-3 is a muscle specific scaffolding protein with both structural and signaling roles. Lack of caveolin-3 expression has been implicated in limb-girdle muscular dystrophy, along with distal myopathy and rippling muscle disease. These diseases are characterized by progressive muscle weakness and muscle wasting. Nicotinic acetylcholine receptor (nAChR) clustering and localization are important for efficient nerve to muscle contractile signal transmission. It is hypothesized that muscle weakness could originate through disrupted nAChR clustering, disrupting the efficiency of signaling from the motorneuron to the muscle. While the molecular mechanisms involved in nAChR clustering remain to be fully defined, we hypothesize caveolin-3 is important for nAChR clustering and overall neuromuscular junction function.Caveolin-3 and the nAChR co-localize and associate evidenced by immunofluorescence and immunoprecipitation. These results were replicated in differentiated wildtype myotubes treated with the nAChR clustering agent, neural agrin. In differentiated caveolin-3 null myotubes, agrin treatment yields a 60% reduction in nAChR clusters as compared to agrin treated wildtype myotubes. Agrin induces nAChR clustering, through activation of muscle specific kinase (MuSK) and downstream through Rac-1 activation. In differentiated wildtype myotubes, Rac-1 activation peaks at 1 hour of agrin treatment, while in differentiated caveolin-3 null myotubes there is dramatically reduced Rac-1 activation upon agrin treatment. Immunoprecipitation of MuSK shows that caveolin-3 and MuSK association peaks at 1 hour of agrin treatment in wildtype cells. This corresponds to the peak of MuSK phosphorylation which also occurs at 1 hour. Agrin induced MuSK phosphorylation was decreased more significantly than the overall decrease in MuSK expression in the caveolin-3 null cells as compared to the wildtype results. These results indicate a role for caveolin-3 in efficient nAChR clustering.Electromyography studies in anesthetized mice indicated lengthened latencies of the muscle action potential in the caveolin-3 null mice as compared to wildtype mice. There were also decreased overall electromyography (EMG) amplitude and EMG area under the curve in caveolin-3 null mice. Comparison of contractile strength in wildtype and caveolin-3 null animals indicated tetanic contractions to be less stable in the caveolin-3 null animals, though there was late potentiation in actual contractile strength.Lack of caveolin-3 affects the neuromuscular junction formation and transmission without affecting overall contractile strength. This research opens a novel view, that correct neuromuscular junction formation and neuromuscular transmission is important in the development of muscular dystrophies.


Share

Citation/Export:
Social Networking:
Share |

Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Hezel, Michael P.mph12@pitt.eduMPH12
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairJiang, Yuyuj5@pitt.eduYUJ5
Committee MemberGalbiati, Ferrucciofeg5@pitt.eduFEG5
Committee MemberPalladino, Michael Jmjp44@pitt.eduMJP44
Committee MemberClemens, Paula Rpclemens@pitt.eduPCLEMENS
Committee MemberDegroat, William Cwcd2@pitt.eduWCD2
Date: 18 December 2009
Date Type: Completion
Defense Date: 25 November 2009
Approval Date: 18 December 2009
Submission Date: 9 December 2009
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Molecular Pharmacology
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: Cav-3; Caveolin-3; electromyography; nAChR; nAChR clustering; neuromuscular junction; neuromuscular transmission; nicotinic acetylcholine receptor
Other ID: http://etd.library.pitt.edu/ETD/available/etd-12092009-131535/, etd-12092009-131535
Date Deposited: 10 Nov 2011 20:09
Last Modified: 15 Nov 2016 13:54
URI: http://d-scholarship.pitt.edu/id/eprint/10253

Metrics

Monthly Views for the past 3 years

Plum Analytics


Actions (login required)

View Item View Item