Limits...
Expression and function of cannabinoid receptors CB1 and CB2 and their cognate cannabinoid ligands in murine embryonic stem cells.

Jiang S, Fu Y, Williams J, Wood J, Pandarinathan L, Avraham S, Makriyannis A, Avraham S, Avraham HK - PLoS ONE (2007)

Bottom Line: We observed significant induction of CB1 and CB2 cannabinoid receptors during the hematopoietic differentiation of murine ES (mES)-derived embryoid bodies.This work has not been addressed previously and yields new information on the function of cannabinoid receptors, CB1 and CB2, as components of a novel pathway regulating murine ES cell differentiation.This study provides insights into cannabinoid system involvement in ES cell survival and hematopoietic differentiation.

View Article: PubMed Central - PubMed

Affiliation: Division of Experimental Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT

Background: Characterization of intrinsic and extrinsic factors regulating the self-renewal/division and differentiation of stem cells is crucial in determining embryonic stem (ES) cell fate. ES cells differentiate into multiple hematopoietic lineages during embryoid body (EB) formation in vitro, which provides an experimental platform to define the molecular mechanisms controlling germ layer fate determination and tissue formation.

Methods and findings: The cannabinoid receptor type 1 (CB1) and cannabinoid receptor type 2 (CB2) are members of the G-protein coupled receptor (GPCR) family, that are activated by endogenous ligands, the endocannabinoids. CB1 receptor expression is abundant in brain while CB2 receptors are mostly expressed in hematopoietic cells. However, the expression and the precise roles of CB1 and CB2 and their cognate ligands in ES cells are not known. We observed significant induction of CB1 and CB2 cannabinoid receptors during the hematopoietic differentiation of murine ES (mES)-derived embryoid bodies. Furthermore, mES cells as well as ES-derived embryoid bodies at days 7 and 14, expressed endocannabinoids, the ligands for both CB1 and CB2. The CB1 and CB2 antagonists (AM251 and AM630, respectively) induced mES cell death, strongly suggesting that endocannabinoids are involved in the survival of mES cells. Treatment of mES cells with the exogenous cannabinoid ligand Delta(9)-THC resulted in the increased hematopoietic differentiation of mES cells, while addition of AM251 or AM630 blocked embryoid body formation derived from the mES cells. In addition, cannabinoid agonists induced the chemotaxis of ES-derived embryoid bodies, which was specifically inhibited by the CB1 and CB2 antagonists.

Conclusions: This work has not been addressed previously and yields new information on the function of cannabinoid receptors, CB1 and CB2, as components of a novel pathway regulating murine ES cell differentiation. This study provides insights into cannabinoid system involvement in ES cell survival and hematopoietic differentiation.

Show MeSH

Related in: MedlinePlus

Effects of cannabinoid ligands on the chemotaxis of ES cells and hematopoietic differentiated ES-derived EB cells (EBs-day 10).Cells were placed in the upper well of the transwell in the presence or absence of specific inhibitors, as indicated. The ligands: 2-AG, Δ9-THC, JWH-015 and SDF-1α were placed in the lower chambers. Data show the mean value of 3 independent experiments (mean±SD). Error bars indicate SD. * P values with asterisk (*, P<0.05) show significant differences from control with media alone.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC1919431&req=5

pone-0000641-g004: Effects of cannabinoid ligands on the chemotaxis of ES cells and hematopoietic differentiated ES-derived EB cells (EBs-day 10).Cells were placed in the upper well of the transwell in the presence or absence of specific inhibitors, as indicated. The ligands: 2-AG, Δ9-THC, JWH-015 and SDF-1α were placed in the lower chambers. Data show the mean value of 3 independent experiments (mean±SD). Error bars indicate SD. * P values with asterisk (*, P<0.05) show significant differences from control with media alone.

Mentions: The chemotaxis assays were performed using Costar Transwells (Corning-Costar, Cambridge, MA). As shown in Figure 4, chemotaxis was observed with differentiated EBs at day 10 in the presence of the Δ9-THC, 2-AG and JWH-015 cannabinoid ligands, while the chemotaxis of undifferentiated ES cells was very low. This chemotaxis was inhibited by the CB1 and CB2 specific inhibitors, AM251 and AM630, respectively. Thus, cannabinoid ligands, such as 2-AG, exogenous Δ9-THC and JWH-015 induce the chemotaxis of hematopoietic differentiated ES-derived EB cells, mediated through both the CB1 and CB2 receptors.


Expression and function of cannabinoid receptors CB1 and CB2 and their cognate cannabinoid ligands in murine embryonic stem cells.

Jiang S, Fu Y, Williams J, Wood J, Pandarinathan L, Avraham S, Makriyannis A, Avraham S, Avraham HK - PLoS ONE (2007)

Effects of cannabinoid ligands on the chemotaxis of ES cells and hematopoietic differentiated ES-derived EB cells (EBs-day 10).Cells were placed in the upper well of the transwell in the presence or absence of specific inhibitors, as indicated. The ligands: 2-AG, Δ9-THC, JWH-015 and SDF-1α were placed in the lower chambers. Data show the mean value of 3 independent experiments (mean±SD). Error bars indicate SD. * P values with asterisk (*, P<0.05) show significant differences from control with media alone.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1919431&req=5

pone-0000641-g004: Effects of cannabinoid ligands on the chemotaxis of ES cells and hematopoietic differentiated ES-derived EB cells (EBs-day 10).Cells were placed in the upper well of the transwell in the presence or absence of specific inhibitors, as indicated. The ligands: 2-AG, Δ9-THC, JWH-015 and SDF-1α were placed in the lower chambers. Data show the mean value of 3 independent experiments (mean±SD). Error bars indicate SD. * P values with asterisk (*, P<0.05) show significant differences from control with media alone.
Mentions: The chemotaxis assays were performed using Costar Transwells (Corning-Costar, Cambridge, MA). As shown in Figure 4, chemotaxis was observed with differentiated EBs at day 10 in the presence of the Δ9-THC, 2-AG and JWH-015 cannabinoid ligands, while the chemotaxis of undifferentiated ES cells was very low. This chemotaxis was inhibited by the CB1 and CB2 specific inhibitors, AM251 and AM630, respectively. Thus, cannabinoid ligands, such as 2-AG, exogenous Δ9-THC and JWH-015 induce the chemotaxis of hematopoietic differentiated ES-derived EB cells, mediated through both the CB1 and CB2 receptors.

Bottom Line: We observed significant induction of CB1 and CB2 cannabinoid receptors during the hematopoietic differentiation of murine ES (mES)-derived embryoid bodies.This work has not been addressed previously and yields new information on the function of cannabinoid receptors, CB1 and CB2, as components of a novel pathway regulating murine ES cell differentiation.This study provides insights into cannabinoid system involvement in ES cell survival and hematopoietic differentiation.

View Article: PubMed Central - PubMed

Affiliation: Division of Experimental Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT

Background: Characterization of intrinsic and extrinsic factors regulating the self-renewal/division and differentiation of stem cells is crucial in determining embryonic stem (ES) cell fate. ES cells differentiate into multiple hematopoietic lineages during embryoid body (EB) formation in vitro, which provides an experimental platform to define the molecular mechanisms controlling germ layer fate determination and tissue formation.

Methods and findings: The cannabinoid receptor type 1 (CB1) and cannabinoid receptor type 2 (CB2) are members of the G-protein coupled receptor (GPCR) family, that are activated by endogenous ligands, the endocannabinoids. CB1 receptor expression is abundant in brain while CB2 receptors are mostly expressed in hematopoietic cells. However, the expression and the precise roles of CB1 and CB2 and their cognate ligands in ES cells are not known. We observed significant induction of CB1 and CB2 cannabinoid receptors during the hematopoietic differentiation of murine ES (mES)-derived embryoid bodies. Furthermore, mES cells as well as ES-derived embryoid bodies at days 7 and 14, expressed endocannabinoids, the ligands for both CB1 and CB2. The CB1 and CB2 antagonists (AM251 and AM630, respectively) induced mES cell death, strongly suggesting that endocannabinoids are involved in the survival of mES cells. Treatment of mES cells with the exogenous cannabinoid ligand Delta(9)-THC resulted in the increased hematopoietic differentiation of mES cells, while addition of AM251 or AM630 blocked embryoid body formation derived from the mES cells. In addition, cannabinoid agonists induced the chemotaxis of ES-derived embryoid bodies, which was specifically inhibited by the CB1 and CB2 antagonists.

Conclusions: This work has not been addressed previously and yields new information on the function of cannabinoid receptors, CB1 and CB2, as components of a novel pathway regulating murine ES cell differentiation. This study provides insights into cannabinoid system involvement in ES cell survival and hematopoietic differentiation.

Show MeSH
Related in: MedlinePlus