Endogenous hemichannels play a role in the release of ATP from Xenopus oocytes.
ATP is an electrically charged molecule that functions both in the supply of energy necessary for cellular activity and as an intercellular signaling molecule. Although controlled ATP secretion occurs via exocytosis of granules and vesicles, in some cells, and under certain conditions, other mechanisms control ATP release. Gap junctions, intercellular channels formed by connexins that link the cytoplasm of two adjacent cells, control the passage of ions and molecules up to 1 kDa. The channel is formed by two moieties called hemichannels, or connexons, and it has been suggested that these may represent an alternative pathway for ATP release. We have investigated the release of ATP through hemichannels from Xenopus oocytes that are formed by Connexin 38 (Cx38), an endogenous, specific type of connexin. These hemichannels generate an inward current that is reversibly activated by calcium-free solution and inhibited by octanol and flufenamic acid. This calcium-sensitive current depends on Cx38 expression: it is decreased in oocytes injected with an antisense oligonucleotide against Cx38 mRNA (ASCx38) and is increased in oocytes overexpressing Cx38. Moreover, the activation of these endogenous connexons also allows transfer of Lucifer Yellow. We have found that the release of ATP is coincident with the opening of hemichannels: it is calcium-sensitive, is inhibited by octanol and flufenamic acid, is inhibited in ASCx38 injected oocytes, and is increased by overexpression of Cx38. Taken together, our results suggest that ATP is released through activated hemichannels in Xenopus oocytes.
Bahima
Laia
L
Laboratory of Molecular and Cellular Neurobiology, Department of Pathology and Experimental Therapeutics, IDIBELL-Medical School, University of Barcelona, Bellvitge Campus, Feixa Llarga s/n, L'Hospitalet de Llobregat, Spain.
Aleu
Jordi
J
Elias
Marc
M
Martín-Satué
Mireia
M
Muhaisen
Ashraf
A
Blasi
Joan
J
Marsal
Jordi
J
Solsona
Carles
C
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D017630:Connexins / Q000235:genetics / Q000378:metabolism; D005260:Female; D005439:Flufenamic Acid / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism* / Q000648:ultrastructure; D007476:Ionophores / Q000494:pharmacology; D007546:Isoquinolines / Q000378:metabolism; D009761:Nystatin / Q000494:pharmacology; D000442:Octanols / Q000494:pharmacology; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D014982:Xenopus laevis
D000893:Anti-Inflammatory Agents; D017630:Connexins; D005456:Fluorescent Dyes; D007476:Ionophores; D007546:Isoquinolines; D000442:Octanols; D016376:Oligonucleotides, Antisense; C102630:connexin 38; D009761:Nystatin; D005439:Flufenamic Acid; D000255:Adenosine Triphosphate; C017475:lucifer yellow; D002118:Calcium