Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2007-1-12
pubmed:abstractText
Members of the Rab3 (A-D) subfamily of small GTPases are believed to play a key role in regulated exocytosis. These proteins share approximately 80% identity at amino acid level. The question of whether isoforms of Rab3 are functionally redundant was the subject of this study. We used RT-PCR analysis, in situ hybridization histochemistry, and confocal microscope-based analysis of immunocytochemistry to show that rat melanotrophs contain about equal amounts of Rab3A and Rab3B transcripts as well as proteins. Therefore, these cells are a suitable model to study the subcellular distribution and the role of these paralogous isoforms in regulated exocytosis. Secretory activity of single cells was monitored with patch-clamp capacitance measurements, and the cytosol was dialyzed with a high-calcium-containing patch pipette solution. Preinjection of antisense oligodeoxyribonucleotides specific to Rab3A, but not to Rab3B, induced a specific blockage of calcium-dependent secretory responses, indicating an exclusive requirement for Rab3A in melanotroph cell-regulated secretion. Although the injection of purified Rab3B protein was ineffective, the injection of recombinant Rab3A proteins into rat melanotrophs revealed that regulated secretion was stimulated by a GTP-bound Rab3A with an intact COOH terminus and inhibited by Rab3AT36N, impaired in GTP binding. These results indicate that Rab3A, but not Rab3B, enhances secretory output from rat melanotrophs and that their function is not redundant.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0363-6143
pubmed:author
pubmed:issnType
Print
pubmed:volume
292
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
C98-105
pubmed:dateRevised
2007-12-3
pubmed:meshHeading
pubmed-meshheading:16822953-Animals, pubmed-meshheading:16822953-Calcium, pubmed-meshheading:16822953-Cells, Cultured, pubmed-meshheading:16822953-Electric Capacitance, pubmed-meshheading:16822953-Exocytosis, pubmed-meshheading:16822953-Immunohistochemistry, pubmed-meshheading:16822953-In Situ Hybridization, pubmed-meshheading:16822953-Injections, pubmed-meshheading:16822953-Melanotrophs, pubmed-meshheading:16822953-Microscopy, Confocal, pubmed-meshheading:16822953-Oligonucleotides, Antisense, pubmed-meshheading:16822953-Patch-Clamp Techniques, pubmed-meshheading:16822953-Pituitary Gland, pubmed-meshheading:16822953-RNA, Messenger, pubmed-meshheading:16822953-Rats, pubmed-meshheading:16822953-Recombinant Proteins, pubmed-meshheading:16822953-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:16822953-Subcellular Fractions, pubmed-meshheading:16822953-Tissue Distribution, pubmed-meshheading:16822953-rab3 GTP-Binding Proteins, pubmed-meshheading:16822953-rab3A GTP-Binding Protein
pubmed:year
2007
pubmed:articleTitle
Distinct role of Rab3A and Rab3B in secretory activity of rat melanotrophs.
pubmed:affiliation
Lab. of Neuroendocrinology-Molecular Cell Physiology, Institute of Pathophysiology, School of Medicine, PO Box 2211, 1001 Ljubljana, Slovenia.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural