Identification of two further gap-junctional proteins, connexin40 and connexin45, in human myometrial smooth muscle cells at term.
The powerful synchronous contractions of the uterus in labor depend on electrical coupling of myometrial smooth muscle cells by gap junctions. In humans and other mammals, gap junctions are scarce in the myometrium of the non-pregnant uterus, but become abundant at term and/or with the onset of labor. Previous work has shown that the gap-junctional protein (connexin) expressed by human myometrial smooth muscle cells is connexin43, the same connexin type that predominates in cardiac muscle. Here we show that two further gap junctional proteins, connexin40 and connexin45, are expressed by the myometrial smooth muscle cells of the human uterus at term. Transcripts encoding the human isoforms of these connexins were demonstrated by Northern blot analysis, and immunoconfocal microscopy enabled precise localization of the corresponding proteins to punctate contact points (i.e., gap junctions) between interacting smooth muscle cells. Double labeling demonstrated that, while some fluorescent spots comprise only connexin43, both connexin40 and connexin45 predominantly colocalize to connexin43-positive fluorescent spots. Triple labeling revealed that where all three connexin types were expressed, they frequently localized to the same gap junction spot. As gap-junctional channels composed of different connexin types have been demonstrated in vitro to have different functional properties, multiple connexin expression may contribute to modulation of gap junction function in human myometrial smooth muscle cells in vivo.
Kilarski
W M
WM
Imperial College School of Medicine, National Heart and Lung Institute, London, UK.
Dupont
E
E
Coppen
S
S
Yeh
H I
HI
Vozzi
C
C
Gourdie
R G
RG
Rezapour
M
M
Ulmsten
U
U
Roomans
G M
GM
Severs
N J
NJ
D015152:Blotting, Northern; D017630:Connexins / Q000032:analysis*; D005260:Female; D006801:Humans; D007743:Labor, Obstetric / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009130:Muscle, Smooth / Q000737:chemistry* / Q000166:cytology; D009215:Myometrium / Q000737:chemistry* / Q000166:cytology; D011247:Pregnancy; D000097003:Gap Junction alpha-5 Protein
D017630:Connexins; D000097003:Gap Junction alpha-5 Protein; C082920:connexin 45