Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
1997-6-3
pubmed:abstractText
We report a new technique for improving the efficiency of single-channel recording in the Xenopus oocyte expression system. Conventional methods of oocyte preparation (adequate for two-electrode voltage-clamp) often leave residual adhesions between the vitelline and plasma membranes. As a result, removal of the vitelline membrane for patch-clamp recording produces microscopic damage to the oocyte plasma membrane that interferes with the formation of high-resistance electrical seals. This problem has been alleviated by a three-phase method of oocyte preparation: (1) a strong initial enzymatic digestion using both collagenase and hyaluronidase, immediately followed by (2) exposure to hypertonic media and mechanical defolliculation, (3) pre-shrinkage and selection of only those oocytes displaying clear separation between vitelline and plasma membranes. Following this selection process, oocytes were returned to solutions of normal osmolarity, incubated overnight at 19 degrees C, and then injected with RNA for expression studies. Comparison of oocytes prepared by different methods indicated that use of these three steps significantly improved the success of high-resistance seal formation, without compromising oocyte survivability or the efficiency of channel expression.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0031-6768
pubmed:author
pubmed:issnType
Print
pubmed:volume
433
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
648-52
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1997
pubmed:articleTitle
Improved preparation of Xenopus oocytes for patch-clamp recording.
pubmed:affiliation
Department of Physiology and Biophysics, Cornell University Medical College, 1300 York Ave., New York NY 10021, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.