RESUMEN
The objective of this study was to document the expression and functional role of BMPs in the placental (caruncle; CAR, cotyledon; COT) during different stages of pregnancy in water buffalo. Samples collected from Early pregnancy 1 (EP1); Early pregnancy 2 (EP2), Mid pregnancy (MP), Late pregnancy (LP) while the third stage of oestrus cycle (NP) was taken as control. Also, the synergistic role of BMP4/BMP7 or combination on mRNA expression of vWF, PCNA, StAR, CYP11A1, 3ßHSD, and BAX were studied in trophoblast cells cultured (TCC) during an early stage. The qPCR and immunoblotting studies revealed that BMP2, BMPR1A, BMPR1B, and BMPR2 mRNA level was significantly (pâ¯<â¯0.05) upregulated during early pregnancy in COTs while in CARs it was significantly upregulated (pâ¯<â¯0.05) during all the stages of pregnancy.BMP4 mRNA level was significantly upregulated (pâ¯<â¯0.05) during early pregnancy in COTs as well as in CARs. BMP6 expression was significantly upregulated (pâ¯<â¯0.05) during early and late stages of pregnancy. BMP7 mRNA level was upregulated (pâ¯<â¯0.05) during the late stage of pregnancy in COTs. At 100â¯ng/ml, the BMP4 maximally stimulated the transcripts of StAR, CYP11A1, and 3ßHSD while BMP7 maximally stimulated the transcripts of 3ßHSD that paralleled with P4 accretion in the media (Pâ¯<â¯0.05). BMP4 as well as BMP7 upregulated the transcripts of PCNA, vWF, and downregulated BAX in the TCC (Pâ¯<â¯0.05). In conclusion, BMPs are expressed in a regulated manner with stage-specific differences in the placenta and promotes the angiogenesis, proliferation, cell survivability, and steroidogenesis thereby regulating placental function in an autocrine/paracrine manner in water buffalo.
Asunto(s)
Proteínas Morfogenéticas Óseas/genética , Búfalos/genética , Regulación del Desarrollo de la Expresión Génica , Placenta/metabolismo , Animales , Proteínas Morfogenéticas Óseas/metabolismo , Supervivencia Celular , Femenino , Neovascularización Fisiológica/genética , Placenta/irrigación sanguínea , Embarazo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de Tiempo , Trofoblastos/citologíaRESUMEN
BMPs and their receptors modulate the granulosa cell (GC) function in the follicle of domestic animals. Since little is known on BMPs in the buffalo, the present study was aimed to investigate the expression of BMP2, 4, 6, 7 and their receptors BMPR1A, BMPR1B, BMPR2 in the GC and theca cells (TC) of ovarian follicles and the role of BMP4 and BMP7 on buffalo GC. Follicles were classified into four groups based on size and E2 level in the follicular fluid as follows: (i) Group1(4-6â¯mm; <0.5â¯ng/mL) (ii) Group 2 (7-9â¯mm; 0.5-5â¯ng/mL) (iii) Group 3 (10-13â¯mm; 5-40â¯ng/mL) and (iv) Group 4 (dominant follicle) (>13â¯mm; >180â¯ng/mL). The results revealed that except BMP6, BMP2, 4 7 and receptors BMPR1A, BMPR1B and BMPR2 showed a minimum of 1.5-2 fold increase in mRNA expression in the GC of dominant follicle as compared to other follicle classes. In the dominant follicle, a two-fold increase in BMP4 and BMP7 expression was observed in the TC. At 100â¯ng/mL, the BMP4 and BMP7 either alone or in combination maximally down-regulated CASPASE3 and stimulated the transcripts of PCNA, FSHR and CYP19A1 that was supported by E2 secretion in the granulosa cell culture suggesting their role in cell survival and E2 production. In conclusion, GC and TC of dominant follicles express BMP 2, 4, 6, 7 and their receptors BMPR1A, BMPR1B and BMPR2. BMP4 and BMP7 stimulate E2 production and promote GC survival.