The variable success of in vitro maturation: can we do better?

The efficiency of in vitro assisted reproductive technologies, consisting of the transfer of embryos obtained in vitro through in vitro maturation, in vitro fertilization and early embryo culture is still limited. The quality of the oocytes is pivotal for assisted reproductive efficiency and the maturation of the oocyte represents the first key limiting step of the in vitro embryo production system. At the time of removal from the antral follicles, the oocyte is still completing the final growth and differentiation steps, needed to provide the so-called developmental competence, i.e. the machinery required to sustain fertilization and embryo development. In mono-ovular species only one oocyte per cycle is available for procreation, therefore the current assisted reproduction techniques strive to overcome this natural boundary. However, the success is still limited and overall the effectiveness does not exceed the efficiency achieved in millions of years of mammalian evolution. One of the problems lies in the intrinsic heterogeneity of the oocytes that are subjected to in vitro maturation and in the lack of dedicated in vitro approaches to finalize the differentiation process. In this review we will try to overview some of the salient aspects of current practices by emphasizing the most critical and fundamental features in oocyte differentiation that should be carefully considered for improving current techniques

The variable success of in vitro maturation: can we do better?

The efficiency of in vitro assisted reproductive technologies, consisting of the transfer of embryos obtained in vitro through in vitro maturation, in vitro fertilization and early embryo culture is still limited. The quality of the oocytes is pivotal for assisted reproductive efficiency and the maturation of the oocyte represents the first key limiting step of the in vitro embryo production system. At the time of removal from the antral follicles, the oocyte is still completing the final growth and differentiation steps, needed to provide the so-called developmental competence, i.e. the machinery required to sustain fertilization and embryo development. In mono-ovular species only one oocyte per cycle is available for procreation, therefore the current assisted reproduction techniques strive to overcome this natural boundary. However, the success is still limited and overall the effectiveness does not exceed the efficiency achieved in millions of years of mammalian evolution. One of the problems lies in the intrinsic heterogeneity of the oocytes that are subjected to in vitro maturation and in the lack of dedicated in vitro approaches to finalize the differentiation process. In this review we will try to overview some of the salient aspects of current practices by emphasizing the most critical and fundamental features in oocyte differentiation that should be carefully considered for improving current techniques(AU)

Compensatory kidney hypertrophy/hyperplasia after nephrectomy in mice: alterations of connexin 43 (Cx43) phosphorylated isoforms

Compensatory kidney hypertrophy/hyperplasia leads to several changes in kidney structure and function, as increased glomeruli filtration. The aim of this study was to evaluate connexin 43 in remnant mouse kidneys after unilateral nephrectomy. The right kidney was surgically removed from BALB/c mice. Groups were euthanized at 24, 48 and 72 hours, and at 7 and 30 days. Kidney sections of the reminiscent kidneys were stained with Periodic Acid/Schiff and additional slides were submitted to BrdU and Cx43 immunohistochemistry. The results demonstrated an increase in kidney weight as early as 24 hours through 30 post-nephrectomy. In addition, BrdU positive epithelial cells increased at 24 and 48 hours post-nephrectomy. Cx43 was detected in the cytoplasm and membrane of epithelial cells and vasculature. Taking into consideration the quantity, intensity and localization of Cx43 immunostaining pattern, we observed that nephrectomized mice presented lower Cx43 expression and a cytoplasmic localization after 24 hours, peaking in 48 hours. Furthermore, western blot revealed that during the first 24 and 48 hours after nephrectomy, PO (unphosphorylated) and P1 (phosphorylated) Cx43 disappeared, and the products of Cx43 degradation were reduced. On the other hand, after 72 hours the PO and P1 state reappeared and the amount of degraded peptides also increased. Seven and thirty days after nephrectomy, a higher intensity of PO and P1 state and a lower P2 (hyperphosphorylated) band were observed. In conclusion, our results suggest that Cx43 phosphorylation results in the retention of Cx43 in cytoplasm and its increased degradation during compensatory renal hyperplasia/hypertrophy.

Compensatory kidney hypertrophy/hyperplasia after nephrectomy in mice: alterations of connexin 43 (Cx43) phosphorylated isoforms

Compensatory kidney hypertrophy/hyperplasia leads to several changes in kidney structure and function, as increased glomeruli filtration. The aim of this study was to evaluate connexin 43 in remnant mouse kidneys after unilateral nephrectomy. The right kidney was surgically removed from BALB/c mice. Groups were euthanized at 24, 48 and 72 hours, and at 7 and 30 days. Kidney sections of the reminiscent kidneys were stained with Periodic Acid/Schiff and additional slides were submitted to BrdU and Cx43 immunohistochemistry. The results demonstrated an increase in kidney weight as early as 24 hours through 30 post-nephrectomy. In addition, BrdU positive epithelial cells increased at 24 and 48 hours post-nephrectomy. Cx43 was detected in the cytoplasm and membrane of epithelial cells and vasculature. Taking into consideration the quantity, intensity and localization of Cx43 immunostaining pattern, we observed that nephrectomized mice presented lower Cx43 expression and a cytoplasmic localization after 24 hours, peaking in 48 hours. Furthermore, western blot revealed that during the first 24 and 48 hours after nephrectomy, P0 (unphosphorylated) and P1 (phosphorylated) Cx43 disappeared, and the products of Cx43 degradation were reduced. On the other hand, after 72 hours the P0 and P1 state reappeared and the amount of degraded peptides also increased. Seven and thirty days after neph