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Estradiol and progesterone regulate the migration of mast cells from the periphery to the uterus and induce their maturation and degranulation.

Jensen F, Woudwyk M, Teles A, Woidacki K, Taran F, Costa S, Malfertheiner SF, Zenclussen AC - PLoS ONE (2010)

Bottom Line: Mast cells (MCs) have long been suspected as important players for implantation based on the fact that their degranulation causes the release of pivotal factors, e.g., histamine, MMPs, tryptase and VEGF, which are known to be involved in the attachment and posterior invasion of the embryo into the uterus.Moreover, MC degranulation correlates with angiogenesis during pregnancy.By using a model of ovariectomized animals, we provide clear evidences that also in vivo, estradiol and progesterone attract MC to the uterus and further provoke their maturation and degranulation.

View Article: PubMed Central - PubMed

Affiliation: Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany. federico.jensen@med.ovgu.de

ABSTRACT

Background: Mast cells (MCs) have long been suspected as important players for implantation based on the fact that their degranulation causes the release of pivotal factors, e.g., histamine, MMPs, tryptase and VEGF, which are known to be involved in the attachment and posterior invasion of the embryo into the uterus. Moreover, MC degranulation correlates with angiogenesis during pregnancy. The number of MCs in the uterus has been shown to fluctuate during menstrual cycle in human and estrus cycle in rat and mouse indicating a hormonal influence on their recruitment from the periphery to the uterus. However, the mechanisms behind MC migration to the uterus are still unknown.

Methodology/principal findings: We first utilized migration assays to show that MCs are able to migrate to the uterus and to the fetal-maternal interface upon up-regulation of the expression of chemokine receptors by hormonal changes. By using a model of ovariectomized animals, we provide clear evidences that also in vivo, estradiol and progesterone attract MC to the uterus and further provoke their maturation and degranulation.

Conclusion/significance: We propose that estradiol and progesterone modulate the migration of MCs from the periphery to the uterus and their degranulation, which may prepare the uterus for implantation.

Show MeSH
BMMCs and HMC-1 express high levels of CD117 as well as estradiol and progesterone receptors.Dot plots of cultured BMMCs (A) or HCM-1 cells (B) stained for CD117/FceRI as and CD117 as analyzed by flow cytometry. (C) BMMCs and HCM-1 cells (D) were stained with toluidine blue and they present typical features of MCs as analyzed by light microscopy using a total augmentation of 1000 X (Zeiss AX 10/Axiovision Rel 4.6). (E) and (F) represent western blots for estrogen receptor (ERα) and beta (ERβ) while (G) represent western blot for progesterone receptor (PR), respectively for HCM-1 (a) and BMMCs cells (b). β-actin served as house keeping gene.
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pone-0014409-g003: BMMCs and HMC-1 express high levels of CD117 as well as estradiol and progesterone receptors.Dot plots of cultured BMMCs (A) or HCM-1 cells (B) stained for CD117/FceRI as and CD117 as analyzed by flow cytometry. (C) BMMCs and HCM-1 cells (D) were stained with toluidine blue and they present typical features of MCs as analyzed by light microscopy using a total augmentation of 1000 X (Zeiss AX 10/Axiovision Rel 4.6). (E) and (F) represent western blots for estrogen receptor (ERα) and beta (ERβ) while (G) represent western blot for progesterone receptor (PR), respectively for HCM-1 (a) and BMMCs cells (b). β-actin served as house keeping gene.

Mentions: It is known that MCs exist in the periphery as precursor cells and migrate to the tissues where they undergo their maturation upon different signals [25]. It has also been reported that the number of MCs oscillate in the uterus of mice throughout estrus cycle [26]. We confirmed that BMMC cultures which contain >97% pure mast cells (MCs) after 5 weeks (Fig. 3A–D) express both estradiol and progesterone receptor (Fig. 3E–G) and that they migrate to uterine cells upon hormones (Fig. 2C). Uterine cells both, in human and mouse release chemokines under hormonal influence throughout menstrual and estrus cycle [15]–[17]., We next analyzed the ability of physiological concentrations of E2 and P4, to modulate the expression of chemokine receptors in the immature human mast cells HMC-1 as well as in mouse bone marrow-derived MCs (BMMCs) as a possible mechanism for MC migration. As previously reported, HMC-1 and BMMCs express both, E2 and P4 receptors [27], [28], [24] (Fig. 3E–G). We then treated HMC-1 cells with 50, 100, 200 and 400 ng/ml of E2 and with 1, 5, 10 and 50 pg/ml of P4 which represent the physiological concentrations of these hormones during the female menstrual cycle in humans. Additionally we treated HMC-1 cells with a combination of both hormones at their most effective concentration (100 pµ/ml-10 ng/ml). We analyzed the expression of CCR4 and CCR5, which represent the most important receptors for chemokines known to be released from the uterus under hormonal influence in humans [15]. As shown in Fig. 4A and 4B, E2 concentrations of 200 and 400 pg/ml or 100 and 400 pg/ml, corresponding to middle cycle and early luteal phase significantly up-regulated the expression of CCR4 and CCR5 respectively on HMC-1 cells after 1 h of treatment. Physiological concentrations of P4 did not affect the expression of these chemokine receptors on HMC-1 cells (data not shown). Furthermore, when HMC-1 cells were treated with a combination of both hormones, a significant up regulation of CCR4 and CCR5 was observed (Fig. 4C, D). In mice, the most important chemokine receptors for the chemokines released from the uterus are CCR3 and CCR5 [16]–[17]. BMMCs were stimulated with physiological concentrations of E2 (10, 15, 20 and 30 pg/ml), P4 (1, 2, 4, 8 ng/ml) or a combination of both (20 pg/ml +4 ng/ml), which mimic the concentrations observed during estrus cycle [29] and after 1 h of treatment the expression of CCR3 and CCR5 in BMMCs was analyzed by flow cytometry. As shown in Fig. 4E–H both, E2 and P4 significantly up-regulated the expression of CCR5 and CCR3. Moreover, a combination of both hormones significantly induced the expression of CCR5 and CCR3 in BMMCs after 1 h of treatment (Fig. 4I–J). Accordingly, CCL5 (RANTES) is able to attract murine MCs (Fig. 4K), which express both CCR3 and CCR5. Thus, CCR4 and CCR5 in humans and CCR3 and CCR5 in mice are capable to mediate MC migration upon hormone signals.


Estradiol and progesterone regulate the migration of mast cells from the periphery to the uterus and induce their maturation and degranulation.

Jensen F, Woudwyk M, Teles A, Woidacki K, Taran F, Costa S, Malfertheiner SF, Zenclussen AC - PLoS ONE (2010)

BMMCs and HMC-1 express high levels of CD117 as well as estradiol and progesterone receptors.Dot plots of cultured BMMCs (A) or HCM-1 cells (B) stained for CD117/FceRI as and CD117 as analyzed by flow cytometry. (C) BMMCs and HCM-1 cells (D) were stained with toluidine blue and they present typical features of MCs as analyzed by light microscopy using a total augmentation of 1000 X (Zeiss AX 10/Axiovision Rel 4.6). (E) and (F) represent western blots for estrogen receptor (ERα) and beta (ERβ) while (G) represent western blot for progesterone receptor (PR), respectively for HCM-1 (a) and BMMCs cells (b). β-actin served as house keeping gene.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3008683&req=5

pone-0014409-g003: BMMCs and HMC-1 express high levels of CD117 as well as estradiol and progesterone receptors.Dot plots of cultured BMMCs (A) or HCM-1 cells (B) stained for CD117/FceRI as and CD117 as analyzed by flow cytometry. (C) BMMCs and HCM-1 cells (D) were stained with toluidine blue and they present typical features of MCs as analyzed by light microscopy using a total augmentation of 1000 X (Zeiss AX 10/Axiovision Rel 4.6). (E) and (F) represent western blots for estrogen receptor (ERα) and beta (ERβ) while (G) represent western blot for progesterone receptor (PR), respectively for HCM-1 (a) and BMMCs cells (b). β-actin served as house keeping gene.
Mentions: It is known that MCs exist in the periphery as precursor cells and migrate to the tissues where they undergo their maturation upon different signals [25]. It has also been reported that the number of MCs oscillate in the uterus of mice throughout estrus cycle [26]. We confirmed that BMMC cultures which contain >97% pure mast cells (MCs) after 5 weeks (Fig. 3A–D) express both estradiol and progesterone receptor (Fig. 3E–G) and that they migrate to uterine cells upon hormones (Fig. 2C). Uterine cells both, in human and mouse release chemokines under hormonal influence throughout menstrual and estrus cycle [15]–[17]., We next analyzed the ability of physiological concentrations of E2 and P4, to modulate the expression of chemokine receptors in the immature human mast cells HMC-1 as well as in mouse bone marrow-derived MCs (BMMCs) as a possible mechanism for MC migration. As previously reported, HMC-1 and BMMCs express both, E2 and P4 receptors [27], [28], [24] (Fig. 3E–G). We then treated HMC-1 cells with 50, 100, 200 and 400 ng/ml of E2 and with 1, 5, 10 and 50 pg/ml of P4 which represent the physiological concentrations of these hormones during the female menstrual cycle in humans. Additionally we treated HMC-1 cells with a combination of both hormones at their most effective concentration (100 pµ/ml-10 ng/ml). We analyzed the expression of CCR4 and CCR5, which represent the most important receptors for chemokines known to be released from the uterus under hormonal influence in humans [15]. As shown in Fig. 4A and 4B, E2 concentrations of 200 and 400 pg/ml or 100 and 400 pg/ml, corresponding to middle cycle and early luteal phase significantly up-regulated the expression of CCR4 and CCR5 respectively on HMC-1 cells after 1 h of treatment. Physiological concentrations of P4 did not affect the expression of these chemokine receptors on HMC-1 cells (data not shown). Furthermore, when HMC-1 cells were treated with a combination of both hormones, a significant up regulation of CCR4 and CCR5 was observed (Fig. 4C, D). In mice, the most important chemokine receptors for the chemokines released from the uterus are CCR3 and CCR5 [16]–[17]. BMMCs were stimulated with physiological concentrations of E2 (10, 15, 20 and 30 pg/ml), P4 (1, 2, 4, 8 ng/ml) or a combination of both (20 pg/ml +4 ng/ml), which mimic the concentrations observed during estrus cycle [29] and after 1 h of treatment the expression of CCR3 and CCR5 in BMMCs was analyzed by flow cytometry. As shown in Fig. 4E–H both, E2 and P4 significantly up-regulated the expression of CCR5 and CCR3. Moreover, a combination of both hormones significantly induced the expression of CCR5 and CCR3 in BMMCs after 1 h of treatment (Fig. 4I–J). Accordingly, CCL5 (RANTES) is able to attract murine MCs (Fig. 4K), which express both CCR3 and CCR5. Thus, CCR4 and CCR5 in humans and CCR3 and CCR5 in mice are capable to mediate MC migration upon hormone signals.

Bottom Line: Mast cells (MCs) have long been suspected as important players for implantation based on the fact that their degranulation causes the release of pivotal factors, e.g., histamine, MMPs, tryptase and VEGF, which are known to be involved in the attachment and posterior invasion of the embryo into the uterus.Moreover, MC degranulation correlates with angiogenesis during pregnancy.By using a model of ovariectomized animals, we provide clear evidences that also in vivo, estradiol and progesterone attract MC to the uterus and further provoke their maturation and degranulation.

View Article: PubMed Central - PubMed

Affiliation: Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany. federico.jensen@med.ovgu.de

ABSTRACT

Background: Mast cells (MCs) have long been suspected as important players for implantation based on the fact that their degranulation causes the release of pivotal factors, e.g., histamine, MMPs, tryptase and VEGF, which are known to be involved in the attachment and posterior invasion of the embryo into the uterus. Moreover, MC degranulation correlates with angiogenesis during pregnancy. The number of MCs in the uterus has been shown to fluctuate during menstrual cycle in human and estrus cycle in rat and mouse indicating a hormonal influence on their recruitment from the periphery to the uterus. However, the mechanisms behind MC migration to the uterus are still unknown.

Methodology/principal findings: We first utilized migration assays to show that MCs are able to migrate to the uterus and to the fetal-maternal interface upon up-regulation of the expression of chemokine receptors by hormonal changes. By using a model of ovariectomized animals, we provide clear evidences that also in vivo, estradiol and progesterone attract MC to the uterus and further provoke their maturation and degranulation.

Conclusion/significance: We propose that estradiol and progesterone modulate the migration of MCs from the periphery to the uterus and their degranulation, which may prepare the uterus for implantation.

Show MeSH