Local VEGF-A blockade modulates the microenvironment of the corneal graft bed.

The microenvironment plays an important role in several immunological processes. Vascular endothelial growth factor-A (VEGF-A) not only regulates angiogenesis, but is known as a modulator of the immune microenvironment. Modulating the site of transplantation might be beneficial for subsequent transplant survival. In this study, we therefore analyzed the effect that a local blockade of VEGF-A in the inflamed cornea as the graft receiving tissue has on the immune system. We used the murine model of suture-induced neovascularization and subsequent high-risk corneal transplantation, which is an optimal model for local drug application. Mice were treated with VEGFR1/R2 trap prior to transplantation. We analyzed corneal gene expression, as well as protein levels in the cornea and serum on the day of transplantation, 2 and 8 weeks later. Local VEGF depletion prior to transplantation increases the expression of pro-inflammatory as well as immune regulatory cytokines only in the corneal microenvironment, but not in the serum. Furthermore, local VEGFR1/R2 trap treatment significantly inhibits the infiltration of CD11c+ dendritic cells into the cornea. Subsequent increased corneal transplantation success was accompanied by a local upregulation of Foxp3 gene expression. This study demonstrates that locally restricted VEGF depletion increases transplantation success by modulating the receiving corneal microenvironment and inducing tolerogenic mechanisms.

Influence of polygamous versus monogamous mating on embryo production in four different strains of mice after superovulatory treatment.

We determined the effect of monogamous or polygamous mating with 2 females on vaginal plug (VP) rate, embryo donors (ED), 2-cell embryo production, and male performance after superovulation of females aging 24d or 45-48d. C57BL/6NCrl (B6N), BALB/cAnCrl (BALB/cN), FVB/NCrl (FVB/N), and Crl:CD1(ICR) (CD-1) females received 5 IU eCG and 5 IU hCG (24d) or 7.5 IU eCG and 7.5 IU hCG (45-48d) 48 h apart. After the hCG injection, females were paired with males, which alternated weekly in monogamous or polygamous mating. Significant differences in the percentage of VP-positive females between monogamous and polygamous mating were observed for B6N (71% vs. 49%), FVB/N (77% vs. 51%), and CD-1 (90% vs. 67%) at 45-48d. BALB/cN and CD-1 showed higher VP rates than B6N and FVB/N. A significantly higher percentage of ED was found for monogamous than for polygamous mating for FVB/N (87% vs. 61%) at 24d and for B6N (91% vs. 53%) and CD-1 (90% vs. 68%) at 45-48d. In all strains of mice and in both age groups, no significant differences were observed in the number of intact 2-cells per VP-positive female, ED or treated female between monogamous and polygamous mating except in the B6N strain where monogamous mating resulted in a significantly higher number of intact 2-cell embryos per treated female than polygamous mating at both ages. The present results imply that polygamous mating can be implemented for 2-cell embryo production in all strains studied except for B6N when all females are euthanized. However, when only VP+ females are sacrificed polygamous mating can be employed for all 4 strains studied.

Maternal obesity attenuates predelivery inflammatory reaction in C57BL/6N mice.

Inflammation and oxidative stress are known to increase before labour. Whether gonadal white adipose tissue (gWAT) participates in this process and whether labour-related processes in placental and adipose tissue are altered in obese women is unknown. In our mouse model, lean mice display elevated placental inflammation and oxidative stress towards the end of pregnancy, accompanied by an increased expression of pro-inflammatory factors in gWAT. Obese mice also display elevated levels of pro-inflammatory factors and oxidative stress in placentas shortly before birth. However, placental infiltration with leukocytes and an increase in gWAT pro-inflammatory factor expression in obese dams are lacking.

Estrous cycle staging before mating led to increased efficiency in the production of pseudopregnant recipients without negatively affecting embryo transfer in mice.

The goal was to increase pseudopregnant mice production by estrous cycle staging by visual examination before pairing and to determine the effect of such pseudopregnant recipients on embryo transfer. To compare methods of estrous cycle staging over 14 days, groups consisted of 10 females in proestrus-estrus and 10 vasectomized males; group 1: only daily visual observation; group 2: daily visual observation and cytological examination on day 1; group 3: daily visual observation and daily cytological examination. The average time to first vaginal plug was 1.8 days in group 1, 2.7 days in group 2, and 3.2 days in group 3, whereas the average time between consecutive vaginal plugs was 9.2 days (group 1), 10 days (group 2), and 9.25 days (group 3). The average time between consecutive estrous cycles was 9.7 days (group 1), 11.8 days (group 2), and 9.4 days (group 3). The congruence between visual and cytological examination in determining proestrus-estrus in group 2 was 100% and that for the four stages in group 3 was 79% with a range of 44% to 100%. From 162 plug-positive females originally selected in proestrus-estrus, 49%, 30%, 19%, and 2% were plug-positive on Day 1, Day 2, Day 3, and Day 4, respectively, showing that pseudopregnant mice production was significantly increased on the first 2 days. From 192 plug-positive females originally selected randomly, these values were 31%, 21%, 35%, 10%, and 3% on d1, d2, d3, d4, and d5, respectively. No significant differences were observed between groups with respect to embryo transfers with fresh or cryopreserved embryos although the number of pups born per litter was higher in group A with fresh (7.57 vs. 6.39) and cryopreserved-thawed embryos (5.0 vs. 4.38). Furthermore, the sex ratio and the genotype of the pups were not significantly affected.