哺乳动物子宫腔胞外囊泡的组分及功能研究进展

胞外囊泡(Extracellular vesicles,EVs)是细胞间通讯的一种媒介,包含两个大亚类-微泡和外泌体。胞外囊泡能选择性地包裹着脂质、蛋白质、mRNAs、miRNAs以及DNA等物质,可以远距离的运输到靶细胞,通过调控靶细胞的转录、翻译等来影响细胞的功能。哺乳动物子宫腔液中含有丰富的胞外囊泡,它们对精子获能,胚胎与母体子宫间的相互识别以及胚胎附植都具有重要作用。本文主要对子宫腔液中胞外囊泡的组分及其在胚胎附植过程中的作用进行综述。     

Perturbations in the uterine luminal fluid composition are detrimental to pregnancy establishment in cattle

Background: A major,unresolved issue is how the uterine microenvironment determines pregnancy success in cattle.Before implantation,conceptus development depends on the uterine secretome(i.e.,histotroph).Despite its pivotal role,little is known about the dynamics of histotroph synthesis and changes in composition throughout the early diestrus and the relevance to pregnancy establishment.We hypothesize that disturbances on histotroph composition affect the establishment of pregnancy.Aim was to disturb histotroph composition at early diestrus and verify the effects on:(Exp.1) timing to restore its composition; and(Exp.2) pregnancy rate after multiple-embryo transfer.Estrous cycle of multiparous Nelore cows were synchronized and estrus was considered d 0(D0) of the experiments.Disturbance was through flushing each uterine horn with 30 m L of DMPBS and collecting the resulting uterine luminal flushing(ULF) on D1; D4; D7; D1 + D4 + D7.Control group remained not-collected.In Exp.1,ULF was collected on D7.5 from all animals and used for quantification of total protein concentration and abundance of albumin.In Exp.2,three in vitro-produced embryos were transferred to the uterine horn ipsilateral to the ovary containing the CL on D7.5 and pregnancy was checked on D25 by ultrasound.Results: In Exp.1,ULF collection on D4 or D7 increased(1.5-to 2.2-folds) the total protein concentration and albumin abundance.ULF collection on D1 did not alter(P 0.10) these endpoints.In Exp.2,ULF collected on D4 or D7 decreased pregnancy rates to approximately half of that measured in the remaining groups.Conclusions: Subtle perturbations imposed to the native intrauterine milieu,such as those caused by a single,low-volume collection of ULF,profoundly disturbs intrauterine composition and pregnancy success.At least 4 d were necessary for the uterus to recover its composition and the functional capacity to carry post-implantation gestation.     

Activity of glycosidases (beta-N-acetyloglucosaminidase, alpha-mannosidase, and beta-galactosidase) in the uterine luminal fluid of cows after multiple ovulation.

This study investigated the activity of beta-N-acetyloglucosaminidase (beta-NAGASE), alpha-mannosidase, and beta-galactosidase in the uterine luminal fluid of cows after superovulation treatment, along with the possible associations between the activity of these 3 glycosidases and the superovulatory response. Embryos and a sample of fluid flushed from each uterine horn were collected on day 7 after artificial insemination (on estrus day 0) from 32 cows in which superovulation was induced with porcine follicle-stimulating hormone. Glycosidase activity was assayed colorimetrically. The cows were classified as to superovulatory response according to the number of corpora lutea per ovary (group 1, 1 to 4; group 2, > 4) and according to the total number of embryos per horn (T1, 0; T2, 1 to 2; T3, 3 to 4; T4, > 4) and the number of transferable embryos per horn (TR1, 0; TR2, 1 to 2; TR3, 3 to 4; TR4, > 4). The mean activity of beta-NAGASE was significantly lower (P < 0.05) in group 2 than in group 1, at 95.99 (standard error 20.43) versus 226.72 (46.77) IU/L. It was also significantly lower (P < 0.01) in group T4 compared with groups T1, T2, and T3, at 50.09 (8.21) versus 129.25 (34.60), 222.27 (62.62), and 290.26 (93.77) IU/L, respectively, as well as in group T1 compared with group T3. There was a positive relationship between beta-NAGASE activity and both the total number of embryos (P = 0.047) and the number of transferable embryos per horn (P = 0.013) when 1 to 4 corpora lutea developed per ipsilateral ovary. No difference in alpha-mannosidase or beta-galactosidase activity was detected among the groups.     

Abdominal ultrasonography in pet guinea pigs

Ultrasonographic examination of 40 pet guinea pigs was performed in the framework of this study. Preparation of the patient for ultrasonography and the examination procedure are described. Normal ultrasonographic findings of the abdominal organs including the liver, spleen, kidney and urinary bladder are presented and illustrated for the first time in this species. Ultrasonography was found to be a useful investigative tool in the diagnosis of abdominal diseases in guinea pigs. In comparison to other diagnostic methods like physical examination including palpation, haematological examination, radiography and diagnostic laparotomy, ultrasonography offers a rapid, non-invasive and reliable means of diagnosing abdominal diseases in this species.     

Immunogenicity of Clostridium septicum in guinea pigs

Five strains of Clostridium septicum were used to prepare bacterins, bacterin-toxoids, toxoid, and combinations of bacterins or bacterin-toxoids. These preparations were tested for immunogenicity in guinea pigs vaccinated subcutaneously with 1.0 ml of product. Usually, a second vaccination was given 21 to 24 days later. The immunity of groups of vaccinated guinea pigs was challenged with as many as 22 strains of C septicum. When challenge exposed with homologous strains at 21 to 24 days after one vaccination or 10 t0 18 days after a second vaccination, 60% to 100% of the guinea pigs in each group survived. Demonstrable cross-protection among strains of C septicum varied from none to 100% protection in vaccinated guinea pigs. A combination of bacterin-toxoid prepared from four selected strains protected 70% to 100% of the vaccinated guinea pigs challenge exposed with 21 strains. Duration-of-immunity studies demonstrated a twofold to fourfold decrease in protection when the vaccination-to-challenge interval was extended an additional 3 weeks. Strains of C septicum do not have an effective common immunogen and the stimulated immunity appears to be of short duration. Antitoxin was demonstrated to be less important than other factors in protecting against C septicum infection.     

Osteodystrophia fibrosa in two guinea pigs.

Two adult guinea pigs were examined because they were lethargic and reluctant to walk. Additionally, I guinea pig had otitis media, and the other had dental malocclusion. Both guinea pigs had been fed a commercially available diet of cereals and pellets enriched with vitamin C and formulated for this species. Radiographically, the guinea pigs had coarse trabecular bone patterns, skeletal deformations, pathologic fractures, and polyarthritic degenerative joint disease. A double cortical line was also evident on several long bones, the pelvis, and the vertebrae. A diagnosis of osteopenia was confirmed by use of dual-energy x-ray absorptiometry. Analysis of a food sample fed to 1 guinea pig revealed calcium and phosphorus contents of 0.524 and 0.425%, respectively (Ca:P ratio, 1.23:1). Microscopic examination of bone tissue from both guinea pigs revealed severe fibrous osteodystrophy. Nutritional secondary hyperparathyroidism caused by calcium-phosphorus imbalance was considered to be the underlying cause of osteodystrophia fibrosa in both guinea pigs.