Expression of prostasin and protease nexin-1 in rhesus monkey (Macaca mulatta) endometrium and placenta during early pregnancy.

Serine proteases have been documented to play key roles in uterine matrix turnover and trophoblastic invasion during implantation. Roles of prostasin serine protease in these processes, however, are currently unclear. The present study was first conducted to investigate the colocalization of prostasin and its cognate serpin, protease nexin-1 (PN-1), in rhesus monkey endometrium and placenta on days 12, 18, and 26 of pregnancy by using in situ hybridization (ISH) and immunohistochemistry. With ISH, expression of prostasin mRNA was intensely localized in the glandular epithelium on days 12 and 18 and in the placental villi, trophoblastic column, trophoblastic shell, and fetal-maternal border on days 18 and 26. With the progress of pregnancy, expression level in the glandular epithelium was significantly decreased, and the accumulation in the placental compartments was further increased. In addition, the stroma and arterioles exhibited modest levels of prostasin signals. However, expression level of PN-1 in these compartments on adjacent sections in the three stages of early pregnancy was weak or below the level of detection. Prostasin protein expression in the endometrium was found to be consistent with the distribution patterns revealed in the ISH experiments. It may be suggested from these results that prostasin is involved in endometrial epithelial morphology establishment, tissue remodeling, and trophoblastic invasion during early pregnancy. The cognate serpin PN-1 was not coordinately expressed along with prostasin, creating a tissue environment favorable for proteolytic activities of prostasin during early pregnancy events.     

Expression of the vascular endothelial growth factor-receptor system in the porcine endometrium throughout the estrous cycle and early pregnancy

The objective of this study was to investigate the protein and mRNA expression of vascular endothelial growth factor (VEGF), VEGFR-1 (fms-like tyrosine kinase, Flt-1) and VEGFR-2 (fetal liver kinase-1/kinase insert domain-containing receptor, Flk-1/KDR) in the endometrium during the estrous cycle and early pregnancy in pigs. The VEGF-receptor system was localized in epithelial and stromal cells, blood vessels, and myometrium. Western blot analysis showed higher levels of VEGF protein during the periovulatory and periimplantation periods (P < 0.001, and P < 0.05, respectively). Constant expression of VEGF mRNA during the cycle and significant upregulation on Days 22-25 of gestation (vs. Days 9-17; P < 0.001) was observed. Stable levels of VEGFR-1 mRNA and protein were detected in the endometrium of cyclic animals. However, higher VEGFR-1 protein expression was found on Days 16-17 of the estrous cycle (P < 0.01) and Days 13-15 of gestation (P < 0.05). Protein expression of VEGFR-2 was elevated on Days 2-4 of the estrous cycle (P < 0.001), but mRNA levels were constant during the cycle. In pregnancy, VEGFR-2 protein expression started to increase after Day 15 (vs. Days 9-12; P < 0.05), but induction of VEGFR-2 mRNA expression occurred earlier on Days 13-15. It appears from the present study that the VEGF-receptor system is regulated in a temporal and spatial manner during the estrous cycle and early pregnancy in pigs. The results suggest that VEGF-A family members are probably involved in appropriate preparation of endometrium for implantation and in vascular events during implantation in pigs.     

VEGF, bFGF, and their receptors in the endometrium of rhesus monkey during menstrual cycle and early pregnancy

A number of cytokines and growth factors are known to modulate proliferation and differentiation of human endometrium. In this study, the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and VEGF receptors, fms-like tyrosine kinase (Flt1) and kinase insert domain-containing region (KDR), and bFGF receptor 1 (Flg) were examined in the endometrium of rhesus monkey on Day 5, 10, 16, 20, 25 of menstrual cycle and on Day 19 of early pregnancy. Western blot analysis showed the specificity of the anti-human antibodies with the monkey tissue. The expression of mRNA and protein of VEGF was correlated with that of its receptor KDR, which was detected in epithelial, vascular, and myometrial cells. The localization of bFGF and its receptor Flg was similar to that of VEGF, except that the Flg was absent in the endothelial cells. Strong expression of VEGF and bFGF in the glandular epithelial cells was observed in the proliferative phase, declined in the secretory phase during the cycle. Stronger staining of these factors was also observed in the decidual cells of the pregnant uterus, as compared with the stromal cells of cycling uterus. No expression of Flt1 was detected in the tissue examined in this study. These data suggest that VEGF, bFGF, and their receptors play important roles in epithelial and stromal development, angiogenesis, and blood vessel function in the endometrium during the menstrual cycle and early pregnancy of the rhesus monkey.     

Expression of vascular endothelial growth factor and its receptors in the porcine corpus luteum during the estrous cycle and early pregnancy

The present study was undertaken to determine the expression of vascular endothelial growth factor (VEGF) and its receptors, fms-like tyrosine kinase (Flt-1) and fetal liver kinase-1/kinase insert domain-containing receptor (Flk-1/KDR), in the porcine corpus luteum (CL) during the estrous cycle and early pregnancy. Immunohistochemical studies localized proteins of VEGF ligand-receptor system in the cytoplasm of luteal cells and in some blood vessels. Western blot analysis revealed significantly higher levels of VEGF protein during early and mid-luteal phase (vs. late luteal phase; P<0.001 and P<0.01, respectively). Quantification of VEGF mRNA in the CL showed increased mRNA levels during entire luteal phase (vs. Days 16-17; P<0.05). Expression of Flt-1 protein remained high during luteal phase (P<0.001), but the mRNA levels tended to increase from the early to the late luteal phase. Elevated protein expression of Flk-1/KDR was found in the mid-luteal phase (vs. Days 16-17; P<0.05). However, induction of Flk-1/KDR mRNA expression occurred earlier, in early luteal phase. The lowest VEGF, Flt-1 and Flk-1/KDR mRNA and protein levels were observed in regressed CL (P<0.001). During pregnancy, VEGF, Flt-1 and Flk-1/KDR mRNA and protein expression was comparable to the mid-luteal phase. In conclusion, the present study has demonstrated dynamic expression of VEGF and its receptors in the porcine CL during the estrous cycle and early pregnancy. These data suggest that the VEGF ligand-receptor system may play an important role in the development and maintenance of the CL in pigs.     

Mandibular growth in the rhesus monkey (Macaca mulatta)

Mandibular growth of 42 rhesus monkeys (Macaca mulatta) was analyzed quantitatively and qualitatively. Four groups of animals were defined according to dentitional age (i.e., infant, juvenile, adolescent, young adult). At each age growth was observed for a 24 week period. Since some animals were observed at more than one stage of development, 57 periods of growth were studied. The growth incremental data were collected by superimposing serial cephalograms on mandibular implants. Growth and remodeling of both the skeletal and dento-alveolar components of the rhesus mandible were greatest in the infant monkeys and were less in successive age groups. Posterior relocation of the ramus was noted in all age groups while bone deposition on the anterior and inferior borders of the mandibular body was greatest in the younger animals. The most pronounced dental changes also occurred in the younger animals while the dentitions of the adolescent and adult animals were generally more stable. This study demonstrates that the rate and direction of normal mandibular growth varies with the age of the animal. Furthermore, mandibular growth is quantified at four defined maturational levels to provide a set of values illustrating normal mandibular growth. These values can also be used as control data for experimental studies.     

Expression of vascular endothelial growth factor receptors and their ligands in rat uterus during the postpartum involution period

Abstract

Vascular endothelial growth factor (VEGF) and its specific receptors, FLt1/fms, Flk1/KDR and FLt4, play important roles in vasculogenesis, and physiological and pathological angiogenesis. Whether angiogenic growth factors are involved in regulating angiogenic processes during the postpartum involution period (PP) of the rat uterus is unknown. We used immunohistochemistry to analyze the expression levels of VEGF, the fms-like tyrosine kinase 1 (FLt1/fms), the kinase insert domain-containing region 1 (Flk1/KDR), Fms-related tyrosine kinase 4 (FLt4) and vascular endothelial growth inhibitor (VEGI) in the rat uterus during the days 1, 3, 5, 10 and 15 of the PP to determine the temporal and spatial expressions of VEGF and its receptors during the PP. Throughout the PP, cytoplasmic and membrane staining of VEGI, VEGF and their receptors were observed in the lumens, crypts and glandular epithelial cells as well as in connective tissue and vascular endothelial and smooth muscle cells in the endometrium. We found that the intensity of the immunoreactions in the endometrium varied with the morphological changes that occurred during involution. Immunoreactions for VEGI, VEGF and their receptor, Flk1/KDR, in the luminal epithelial cells were stronger than those in the glandular epithelial and stromal cells, particularly during PP 1, 3 and 5, which suggests that these peptides may contribute to re-epithelialization of the endometrium. On the other hand, Flt1/fms immunoreactivity was strong mainly in the stromal cells during the PP. The presence of VEGF and its receptors (FLt1/fms, Flk1/KDR, FLt4) in the stromal cells and blood vessels during the PP suggests that they may contribute to regulating stromal repair and angiogenesis in the involuting uterus of the rat.     

Expression of vascular endothelial growth factor (VEGF) and its cognate receptors in human pheochromocytomas

Pheochromocytomas are well-vascularized tumors, suggesting that a potent angiogenic factor may be involved in the mechanism of their formation. As vascular endothelial growth factor (VEGF) is a potent mitogen for vascular endothelial cells, here we have investigated the mRNA and protein expression of VEGF and the mRNA expression of its two receptors (Flt-1 and Flk-1/KDR) in pheochromocytomas tissue. An increase in VEGF mRNA (mainly isoforms VEGF(121) and VEGF(165)) and in VEGF protein expression were observed by semi-quantitative RT-PCR and Western blot, respectively, compared to normal adrenomedullary tissue. Flk-1/KDR, and Flt-1 levels of mRNA were also increased markedly in tumors and correlated with levels of VEGF mRNA. Therefore, we speculate that upregulation of VEGF expression and its receptors might be important in the pathogenesis of pheochromocytomas.     

血管内皮生长因子及其受体与Cyclin E-CDK2在肝癌发生发展过程中的表达变化

目的通过观察血管内皮生长因子(VEGF)及其受体和细胞周期蛋白E(Cyclin E)在肝癌模型大鼠肝脏中表达情况,探讨VEGF与细胞周期相关蛋白在肝癌发生发展过程中的作用。方法建立诱发性肝癌模型,采用酶联免疫吸附试验检测血清中VEGF量的变化,免疫组化技术检测VEGFR1、Cyclin E和细胞周期蛋白依赖性激酶(CDK2)的表达情况。结果血清中的VEGF含量在对照组中最低,在实验组中逐渐增多,以癌变期含量最高。VEGFR1、Cyclin E和CDK2蛋白表达的平均光密度值均随着肝癌的发生发展有增高的趋势,大鼠血清中的VEGF量与肝脏组织中VEGFR1、Cyclin E和CDK2蛋白表达的平均光密度值随着肝癌的发生发展呈正相关(r=0.834,F=42.1274,P<0.05)。结论 VEGF及其受体VEGFR1在肝癌发生发展中异常表达,促进肝癌的发生发展,可能与Cyclin E、CDK2细胞周期蛋白异常表达有关。     

恒河猴胚胎浅表型植入机理的探讨

为揭示恒河猴胚胎浅表型植入的分子机理,实验采用免疫组化和原位杂交的方法研究了粘附侵润相关分子在妊娠恒河猴母胎界面的表达模式,发现层粘连蛋白(LN)、Ⅳ型胶原(ColⅣ)和基质金属蛋白酶-2(MMP-2)在滋养层细胞柱的远端开始表达;整合素α1β1从柱的近端到远端表达逐渐升高,但在滋养层细胞鞘中上述分子的表达均降低;大量侵润母体血管的滋养层细胞高水平表达上述分子。此外,植入早期上皮斑和蜕膜基质细胞高表达MMPs组织抑制因子-2(TIMP-2)。结果提示,滋养层细胞鞘中α1β1、LN、MMP-2表达水平的降低及上皮斑、蜕膜和滋养层细胞中TIMP-2的高表达可能是导致浅表型植入形成的分子基础。     

Corneal biometrics of the rhesus monkey (Macaca mulatta)

Keratometry, retinoscopy, horizontal diameter, and pachymetry were measured in adult and infant rhesus monkeys (Macaca mulatta). The adult monkeys had a mean corneal thickness of 0.47 +/- 0.03 mm (+/- standard deviation), corneal diameters of 10.6 +/- 0.5 mm, keratometry values of 51.90 +/- 1.61 diopters, and subjective refractions of 0.48 +/- 2.36 diopters. Female adult monkeys had smaller corneal diameters by 0.06 mm (P = .02) and steeper corneas by 0.77 diopters (P = .04). Infant female monkeys also had steeper corneas by 2.37 diopters (P = .05). No significant differences were found between right and left eyes in either adult or infant monkeys.