Gap junctional intercellular communication is required to maintain embryonic stem cells in a non-differentiated and proliferative state.
Pluripotent embryonic stem (ES) cells are capable of maintaining a self-renewal state and have the potential to differentiate into derivatives of all three embryonic germ layers. Despite their importance in cell therapy and developmental biology, the mechanisms whereby ES cells remain in a proliferative and pluripotent state are still not fully understood. Here we establish a critical role of gap junctional intercellular communication (GJIC) and connexin43 (Cx43) in both processes. Pharmacological blockers of GJIC and Cx43 down-regulation by small interfering RNA (siRNA) caused a profound inhibitory effect on GJIC, as evidenced by experiments of fluorescence recovery after photobleaching. This deficient intercellular communication in ES cells induced a loss of their pluripotent state, which was manifested in morphological changes, a decrease in alkaline phosphatase activity, Oct-3/4 and Nanog expression, as well as an up-regulation of several differentiation markers. A decrease in the proliferation rate was also detected. Under these conditions, the formation of embryoid bodies from mouse ES cells was impaired, although this inhibition was reversible upon restoration of GJIC. Our findings define a major function of GJIC in the regulation of self-renewal and maintenance of pluripotency in ES cells.
Todorova
Mariana G
MG
Institute of Bioengineering, Miguel Hernandez University, Elche, Alicante, Spain.
Soria
Bernat
B
Quesada
Ivan
I
D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D036681:Fluorescence Recovery After Photobleaching; D017629:Gap Junctions / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D020868:Gene Silencing; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis / Q000378:metabolism; D018398:Homeodomain Proteins / Q000032:analysis / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D000071317:Nanog Homeobox Protein; D016900:Neurofilament Proteins / Q000032:analysis / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000032:analysis / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry / Q000494:pharmacology
D018031:Connexin 43; D004268:DNA-Binding Proteins; D005904:Glial Fibrillary Acidic Protein; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D016900:Neurofilament Proteins; D050814:Octamer Transcription Factor-3; D034741:RNA, Small Interfering; C051774:neurofilament protein H; D000469:Alkaline Phosphatase