Role of the Major Ecto-Phosphoprotein in Sperm Flagellar Motility Using a Cell Electroporation Method
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Role of the Major Ecto-Phosphoprotein in
Sperm Flagellar Motility Using a Cell
Electroporation Method
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| Creator |
Maiti, Arunima
Mishra, K P Majumder, Gopal Chandra |
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| Subject |
Cell Biology & Physiology
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| Description |
Previous studies from our laboratory
have identified MPS, a 100-kDa protein, as the
major phosphoprotein substrate of caprine sperm ectocyclic
AMP independent protein kinase. In this study
the isolated 32P-labelled MPS has been incorporated
into mature caprine (Capra indicus) cauda-epididymal
spermatozoa with the help of cell electroporation
technique to investigate the effect of MPS on sperm
flagellar motility. The optimum conditions for electroporation
of sperm cells consisted of exposure of 0.2 ml
of sperm cells (2�108/ml) to external electric field of
intensity 1.5 kV/cm and capacitation of 25 mF at 48C
and post-pulse incubation at 378C for 1 hr. when
nearly 50% of the cells lost motility. Scanning electron
micrographs (SEM) demonstrate the formation of
micro-pores and local osmotic swelling in the electroporated
spermatozoa. MPS incorporation was maximal
when its concentration was 30 mg/ml (300 pmol) in the
medium and when the post-pulse incubation time was
60 min. At maximum (75%) MPS incorporation, total
and forward motility increments were also maximum:
34% (P<0.01) and 32% (P<0.01), respectively.
The subcellular fractionation data show that major
portion of the introduced MPS was bound to the
plasma-membrane of spermatozoa. The 32P-labelled
electroporesed intact spermatozoa lost radioactivity
due to the action of the endogenous ecto-phosphoprotein
phosphatase. Therefore MPS is primarily localised
on the sperm external surface leaving its phosphate
group(s) oriented in the extracellular medium. The data
provided further evidence to strengthen the view that
MPS is an ecto-phosphoprotein and that it plays an
important role in the regulation of sperm flagellar
motility.
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| Date |
2008
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/200/1/MOLECULAR_REPRODUCTION_AND_DEVELOPMENT%2C75_(7)%2C_1185%2D1195%2C2008_[53].pdf
Maiti, Arunima and Mishra, K P and Majumder, Gopal Chandra (2008) Role of the Major Ecto-Phosphoprotein in Sperm Flagellar Motility Using a Cell Electroporation Method. Molecular Reproduction and Development, 75 (7). pp. 1185-1195. |
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| Relation |
http://dx.doi.org/10.1002/mrd.20854
http://www.eprints.iicb.res.in/200/ |
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