For MIF staining, sections (4-im thickness) of paraffin-embedded placenta tissues were cut, deparaffinized, rehydrated, and washed in Tris-buffered saline (TBS; 20 mM Tris-HCl [pH 7.6], 150 mM NaCl). TBS was used for all subsequent washes and for dilution of the antibody. Antigen retrieval was carried out by incubating sections in 10 mM sodium citrate buffer (pH 6.0) in a microwave oven at 750 W for 5 min. For CSF1 immunostaining, frozen tissues were cut (6-im thickness) and fixed in ice-cold acetone.
Anti-human MIF and CSF1 goat polyclonal antibodies were purchased from R&D Systems (Abingdon, UK) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively. Horseradish peroxidase-conjugated rabbit anti-goat antibody was obtained from Calbiochem (San Diego, CA). All the chemicals were of analytical grade (Sigma Chemical Co., St. Louis, MO).
In keeping with its known immunomodulatory functions, we have proposed the involvement of MIF in regulating macrophage accumulation in the pregnant endometrium.
The current study evaluated the involvement of CSF1 and MIF in the recruitment and maintenance of macrophages in human decidua. After demonstrating the presence of these two potentially antagonistic chemokines by immunohistochemical staining of sections of term decidua, we sought to elucidate the mechanisms underlying decidual CSF1 and MIF expression.
These mice are depleted of circulating monocytes and tissue macrophages in several organs, including the uterus. They experience severely reduced fertility, lower implantation rates, and greater embryonic wastage compared with wild-type females. Paradoxically, excess decidual macrophage infiltration has been linked to preeclampsia and intrauterine growth restriction in humans.
The early pregnant human endometrium (decidua) contains a diverse population of leukocytes that mediate innate immunity, including uterine natural killer (uNK) cells (7075%) and macrophages (20-25%). In contrast, T and B lymphocytes, which mediate adaptive immunity, account for 10% and between 1% and 3 % of the white cell population, respectively. In recent years, research has focused on the role played by uNK in placentation, with far less attention being directed towards decidual macrophages.
In addition, aromatase activity may reflect the response of Lhb in the pituitary in vivo. The effects of GnRH and its various analogs on gonadotropin have been extensively studied in mammals and teleosts. It has been determined that GnRH is secreted from the hypothalamus in a pulsatile fashion, which parallels the pulsatile release of LH and FSH, and that a feedback regulation exists between them. It has also been demonstrated that GnRH is capable of stimulating cga, fshb, and lhb mRNA levels in the goldfish pituitary following 24 h of treatment in vivo.
Our further study of the effects of rgLh (0.5, 2.5, 5, 10 ig) on T and E2 secretion with different treatment time (2, 6, 12, and 24 h) suggests that rgLh can stimulate sex steroid hormone secretion in a time-dependent manner.
Gonadotropin secretion in vertebrates is mainly regulated by the brain-pituitary-gonadal axis. It is produced in the pituitary by the stimulation of GnRH produced in the hypothalamus and acts on the gonad to produce androgens and estrogens.
In vitro experiments on the vitellogenesis of common carp, tuna, and chum salmon show that Fsh and Lh have equivalent function in stimulating ovarian steroid production. Experiments have shown that sex steroids differentially regulate gonadotropin gene expression in the pituitary of goldfish: a strong in vivo inhibitory effect on fshb mRNA production, but a weak stimulatory effect on lhb in sexually immature and recrudescent fish. T treatment in vitro does not significantly decrease fshb mRNA levels, but increases that of lhb only in the cells of immature fish.
Baculoviruses offer many advantages over other expression vector systems, including safety, ease of scale-up, high levels of recombinant gene expression, and accuracy. In the present study, Lh of the orange-spotted grouper was synthesized using the Bac-to-Bac baculovirus expression system.
It has been accepted that FSH is involved in the control of puberty and gametogenesis and LH regulates the maturation of the ovary and spermatogenesis in mammals.