In vivo oocyte IGF-1 priming increases inner cell mass proliferation of in vitro-formed bovine blastocysts

Studies addressing the effects of supraphysiological levels of IGF-1 on oocyte developmental competence are relevant for unravelling conditions resulting in high bioavailability of IGF-1, such as the polycystic ovary syndrome (PCOS). This study investigated the effects of supraphysiological levels of IGF-1 during in vivo folliculogenesis on the morula-blastocyst transition in bovine embryos. Compacted morulae were non-surgically collected and frozen for subsequent mRNA expression analysis (IGF1R, IGBP3, TP53, AKT1, SLC2A1, SLC2A3, and SLC2A8), or underwent confocal microscopy analysis for protein localization (IGF1R and TP53), or were cultured in vitro for 24 h. In vitro-formed blastocysts were subjected to differential cell staining. The mRNA expression of SLC2A8 was higher in morulae collected from cows treated with IGF-1. Both IGF1R and TP53 protein were present in the plasma membrane and cytoplasm. IGF-1 treatment did not affect protein localization of both IGF1R and TP53. In vitro-formed blastocysts derived from morulae recovered from IGF-1-treated cows displayed a higher number of cells in the inner cell mass (ICM). Total cell number (TCN) of in vitro-formed blastocysts was not affected. A higher mean ICM/TCN proportion was observed in in vitro-formed blastocysts derived from morulae collected from cows treated with IGF-1. The percentage of in vitro-formed blastocysts displaying a low ICM/TCN proportion was decreased by IGF-1 treatment. In vitro-formed blastocysts with a high ICM/TCN proportion were only detected in IGF-1 treated cows. Results show that even a short in vivo exposure of oocytes to a supraphysiological IGF-1 microenvironment can increase ICM cell proliferation in vitro during the morula to blastocyst transition.     

Influence of carbamoylation on some analytical properties of basic polypeptides

The effect of carbamoylation on the assay or identification of histones and polylysine was investigated. Incubation with sodium cyanate decreased the positive charge on these polypeptides as judged by changes in the binding of methyl orange or the electrophoretic mobility. Histones in chromatin appeared less accessible to carbamoylation than isolated histones. Carbamoylation of proteins under conditions in which there was little or no effect on the Lowry procedure could affect their assay by methods utilizing metachromasia with Coomassie Blue G. The Bradford assay has low sensitivity for Hl histone and polylysine but this can be increased by preincubation with sodium cyanate. More extensive carbamoylation of polylysine caused decreased sensitivity which was the only response seen with core nucleosomal histones and bovine serum albumin when preincubated with sodium cyanate. It was concluded that the sensitivity for Hl histone and polylysine in assays dependent on metachromasia with Coomassie Blue G may be changed by factors which decrease the positive charge on these polypeptides.     

Androgen regulation of the motor copulatory pattern in the male New Zealand white rabbit

The effect of castration and testosterone propionate (TP) treatment on the motor copulatory pattern was studied by an accelerometric technique in five sexually experienced New Zealand white male rabbits. This technique permits the oscillographic recording of thrusting frequency, rhythmicity, and amplitude of the pelvic movements occurring during copulation. Castration resulted in a marked decrease in sexual activity in all rabbits. Mounting, including occasional intromissions, was retained by four of the rabbits for periods ranging from 2 to 15 weeks. Castration did not affect mount duration, but decreased strength and frequency of pelvic thrusting. Diminution in the frequency of pelvic thrusting was mainly due to intercalation of pauses within the mounting trains. The change in the rhythm of pelvic thrusting was related to the failure of most mounts performed by castrated rabbits to stimulate lordosis in the female. Testosterone propionate (TP, 10 mg daily for 15 days) restored mounting activity and increased strength and frequency of pelvic thrusting in all rabbits. It is concluded that TP, besides stimulating sexual motivation, regulates the vigor and rhythm of pelvic movements during copulation in the rabbit.     

Perfusion redistribution after alveolar flooding: vasoconstriction vs. vascular compression

We compared the effects of left caudal lobe (LCL) alveolar hypoxia on regional pulmonary blood flow (PBF) with the effects due to alveolar edema induced by plasma instilled directly into the LCL airways of 16 dogs. Regional measurements were made with positron emission tomography. After hypoxic ventilation of the LCL (n = 11), the LCL-to-right caudal lobe (L/R) PBF ratio fell from 0.94 +/- 0.21 during 100% oxygen ventilation of the LCL to 0.46 +/- 0.21 (P less than 0.05). After instillation of either isooncotic (n = 5) or hypooncotic plasma (n = 3) into the LCL, the L/R PBF ratio was similar to that during LCL hypoxia (0.50 +/- 0.27 and 0.64 +/- 0.10, respectively, P less than 0.05 compared with 100% oxygen ventilation of the LCL before plasma instillation). The changes in regional PBF due to LCL hypoxia and plasma instillation could be completely prevented (n = 8) by the prior administration of a single dose of endotoxin (15 micrograms/kg iv). In contrast to previous work, these results indicate that perfusion redistribution occurs regardless of the oncotic state of alveolar edema. More importantly, any change that does occur in regional PBF can be completely prevented by blocking regional vasoconstriction, indicating that mechanical compression cannot be the major factor determining the regional response of PBF to alveolar flooding.