Activin βA subunit, follistatin and follistatin-like 3 are expressed in the endometrium of ovariectomized rats and regulated by estrogen replacement

Abstract

Activin A is a growth factor expressed in the endometrium, where it modulates tissue remodeling and enhances decidualization. The effects of activin A are counteracted by two binding proteins, namely follistatin and follistatin-like 3 (FSTL3). We have evaluated the effects of estrogen and progestin on the endometrial expression of activin betaA subunit, follistatin and FSTL3 in ovariectomized rats. Adult female Wistar rats (n = 21) were ovariectomized and received one week later a single dose of estradiol benzoate (1.5 mg/kg body weight, i.m. injection), either alone (n = 7) or associated with depot medroxyprogesterone acetate (3 mg/kg body weight, i.m. injection, n = 7), or oil vehicle (control group, n = 7). One week later, activin betaA subunit mRNA levels had increased significantly in the uteri of rats treated with estradiol alone (7.4 fold increase over controls, P < 0.05) and to the same extent in rats receiving estradiol plus medroxyprogesterone (6.1 fold increase over controls, P < 0.05). This was accompanied by increase of betaA subunit immunostaining in estradiol and estroprogestin treated rats, which was noted only in the surface endometrial epithelium. Follistatin mRNA expression, conversely, showed a significant decrease in the groups treated with estrogen alone and estrogen plus progestin (P < 0.05), and follistatin immunostaining in the glandular epithelium was weaker in estradiol and estroprogestin-treated rats compared to controls. FSTL3 expression was similar in the 3 groups. In conclusion, the expression of activin betaA subunit increases and that of follistatin decreases following estrogen replacement in the endometrium of ovariectomized rats, and these effects are not further altered by the addition of progestin.

Effect of physical training on liver expression of activin A and follistatin in a nonalcoholic fatty liver disease model in rats

“…For real-time PCR, 20 ng cDNA and 0.4 µM of each primer were used in a 25 µL reaction volume containing SYBR Green PCR Master Mix (Applied Biosystems Inc., USA). The primer sequences used were: beta-A activin (forward primer: 5′-ATGGACCTAACTCTCAGCCAGA-3′; reverse primer: 5′-CTCTCCCCCTTCAAGCCCAT-3′); follistatin (forward primer: 5′-GGCGTACTGCTTGAAGTGAA-3′; reverse primer: 5′-GGGAAGCTGTAGTCCTGGTC-3′); GAPDH (forward primer: 5′-GATGCTGGTGCTGAGTATGTCG-3′; reverse primer: 5′-GTGGTGCAGGATGCATTGCTGA-3′) (18,19). In addition, a rat liver cDNA standard was run in duplicate for every plate to produce a standard curve for quantification.…”

Differential expression of follistatin and FLRG in human breast proliferative disorders

“…The sections were incubated with the primary antibodies in 0.1% BSA/PBS at 4°C overnight at a final dilution of 1:200. Primary polyclonal antibodies raised against human antigens used in the present experiment were: rabbit anti-FST kindly donated by Dr. Wylie Vale (Salk Institute, La Jolla, USA) as previously described [21-23] and rabbit anti-FLRG kindly donated by Dr Véronique Maguer-Satta (Centre Léon Bérard, Lyon, France) [21,22,24,25]. Subsequently, biotinylated secondary antibodies were added to the sections during 30 min followed by peroxidase streptavidin incubation in ABC reagent (Vectastain Elite Universal Kit - Vector Laboratories, Burlingame CA, EUA).…”

Expression of myostatin, myostatin receptors and follistatin in diabetic rats submitted to exercise

“…The primer sequences used were as follows: for MSTN, 5′-CAA ACA GCC TGA ATC CAA CTT AG-3′ (forward) and 5′-CCG TGA GGG GGT AGC GAC AG-3′ (reverse); for ActRIIB, 5′-CAG GTTGGC ACC AGA CGG TAC-3′ (forward) and 5′-TCG ATG GTC ACG CAG AGC TGG-3′ (reverse); for FS, 5′-GGC GTA CTG CTT GAA GTG AA-3′ (forward) and 5′-GGG AAG CTG TAG TCC TGG TC-3′ (reverse); for ALK4, 5′-CAC TGA CAC CAT AGA CAT TGC T-3′ (forward) and 5′-GGC AGA GGA ACA GCT TAA ATC-3′ (reverse). [19][20][21][22]…”