Biology Reference
In-Depth Information
4
∗
P-UAEC
NP-UAEC
3
2
∗
1
∗
∗
∗
0
Treatment
/Time (min)
Fig. 11.4
Immunoprecipitation evidence for IP3R2 interaction with TRPC3 in NP- and P-UAEC.
P- and NP-UAEC were stimulated with 100
M ATP for 8 or 19 min, after which immunopre-
cipitation of IP3R2 was performed. Western blots of recovered protein were probed for TRPC3
and IP3R2. The graph depicts the ratio of TRPC3 in the elutions to TRPC3 in the whole cell
lysates, representing fold enrichment. Data are expressed as mean
μ
0.05.
∗
denotes
significant difference from unstimulated control. IgG control values are also shown to verify the
specificity of the IP3R2 antisera. All data are modified from that published in [12]
±
SE.
n
=
4.
P
=
association between IP3R2 and TRPC3 was observed by immunoprecipitation in
NP- and P-UAEC and, more importantly, this association was only increased after
ATP stimulation of P-UAEC but NOT in the case of NP-UAEC (Fig. 11.4) [12].
These observations confirmed for the first time the direct interaction of IP3R2 with
TRPC3 in UAEC and also showed that this specific interaction was selectively
enhanced by pregnancy. The key to further understanding the pregnancy specific
difference in CCE was in identifying at what level this association was being facil-
itated in P-UAEC that was comparatively lacking in NP-UAEC. The answer to this
question has since been found to be in the pregnancy specific control of cell-cell
communication.
11.8 Evidence that UA Endothelial Cells Communicate
and Coordinate Their Function Via Gap Junctions
The first study that suggested cell-cell communication occurred in UAEC came
from a comparison of the signal responses of UAEC at passage 4 to those of small
groups of freshly isolated cells (initially performed to ensure the pregnancy spe-
cific changes in cell signaling we observed were a true reflection of events in vivo
[9]). Studies of the enhancement of ERK phosphorylation and the more sustained
nature of the [Ca
2+
]i responses in cells freshly isolated from pregnant ewes were
all consistently observed as previously reported for cultured cells [3, 6]. However,
Search WWH ::
Custom Search