Morphological and Gene Expression Changes in Cattle Embryos from Hatched Blastocyst to Early Gastrulation Stages after Transfer of In Vitro Produced Embryos

A detailed morphological staging system for cattle embryos at stages following blastocyst hatching and preceding gastrulation is presented here together with spatiotemporal mapping of gene expression for BMP4, BRACHYURY, CERBERUS1 (CER1), CRIPTO, EOMESODERMIN, FURIN and NODAL. Five stages are defined based on distinct developmental events. The first of these is the differentiation of the visceral hypoblast underlying the epiblast, from the parietal hypoblast underlying the mural trophoblast. The second concerns the formation of an asymmetrically positioned, morphologically recognisable region within the visceral hypoblast that is marked by the presence of CER1 and absence of BMP4 expression. We have termed this the anterior visceral hypoblast or AVH. Intra-epiblast cavity formation and the disappearance of the polar trophoblast overlying the epiblast (Rauber’s layer) have been mapped in relation to AVH formation. The third chronological event involves the transition of the epiblast into the embryonic ectoderm with concomitant onset of posterior NODAL, EOMES and BRACHYURY expression. Lastly, gastrulation commences as the posterior medial embryonic ectoderm layer thickens to form the primitive streak and cells ingress between the embryonic ectoderm and hypoblast. At this stage a novel domain of CER1 expression is seen whereas the AVH disappears. Comparison with the mouse reveals that while gene expression patterns at the onset of gastrulation are well conserved, asymmetry establishment, which relies on extraembryonic tissues such as the hypoblast and trophoblast, has diverged in terms of both gene expression and morphology.     

The total burden of rare,non-synonymous exome genetic variants is not associated with childhood or late-life cognitive ability

Human cognitive ability shows consistent, positive associations with fitness components across the life-course. Underlying genetic variation should therefore be depleted by selection, which is not observed. Genetic variation in general cognitive ability (intelligence) could be maintained by a mutation–selection balance, with rare variants contributing to its genetic architecture. This study examines the association between the total number of rare stop-gain/loss, splice and missense exonic variants and cognitive ability in childhood and old age in the same individuals. Exome array data were obtained in the Lothian Birth Cohorts of 1921 and 1936 (combined N = 1596). General cognitive ability was assessed at age 11 years and in late life (79 and 70 years, respectively) and was modelled against the total number of stop-gain/loss, splice, and missense exonic variants, with minor allele frequency less than or equal to 0.01, using linear regression adjusted for age and sex. In both cohorts and in both the childhood and late-life models, there were no significant associations between rare variant burden in the exome and cognitive ability that survived correction for multiple testing. Contrary to our a priori hypothesis, we observed no evidence for an association between the total number of rare exonic variants and either childhood cognitive ability or late-life cognitive ability.     

Standardizing Immunohistochemistry: A New Reference Control for Detecting Staining Problems

A new standardized immunohistochemistry (IHC) control for breast cancer testing comprises formalin-fixed human epidermal growth factor receptor 2, estrogen receptor, or progesterone receptor peptide antigens covalently attached to 8-µm glass beads. The antigen-coated beads are suspended in a liquid matrix that hardens upon pipetting onto a glass microscope slide. The antigen-coated beads remain in place through deparaffinization, antigen retrieval, and immunostaining. The intensity of the beads’ stain provides feedback regarding the efficacy of both antigen retrieval and immunostaining. As a first report, we tested the sensitivity and specificity of the new IHC controls (“IHControls”). To evaluate sensitivity, various staining problems were simulated. IHControls detected primary and secondary reagent degradation similarly to tissue controls. This first group of IHControls behaved similarly to tissue controls expressing high concentrations of the antigen. The IHControls were also able to detect aberrations in antigen retrieval, as simulated by sub-optimal times or temperatures. Specificity testing revealed that each antigen-coated bead was specific for its cognate IHC test antibody. The data support the conclusion that, like tissue controls, IHControls are capable of verifying the analytic components of an immunohistochemical stain. Unlike tissue controls, IHControls are prepared in large bulk lots, fostering day-to-day reproducibility that can be standardized across laboratories.     

Consort Intrusion and Female Mate Choice in Japanese Macaques (Macaca fuscata)

Consort intrusion behavior has been proposed to largely function as a means of preventing or suppressing the reproductive activity of subordinate males. In the Japanese macaques of Arashiyama B troop this behavior was found to be ineffective in influencing female mate choice and affecting differential reproduction. Mainly directed towards the female of the pair intrusion resembled sexual solicitation and appeared to be an extension of consort behavior. Although the intruder male was almost always dominant over the other male, intrusion failed to provide priority of access to the estrous female as she most frequently chose to remain with her current partner. Copulations which were intruded upon were as likely to end in ejaculation as those which were undisturbed. Only a small proportion of intruder male-female pairs mated. Pairs that did mate were likely to have been in consort before or during the period in which intrusion occurred. No significant correlation was found between male rank order and number of possible females impregnated. Individual young, middle to lower ranking males were estimated to have been reproductively the most successful.     

Dominance among male chimpanzees in the Mahale Mountains National Park,Tanzania: A preliminary study

Dominance relationships among male chimpanzees in the Mahale Mountains National Park, Tanzania, were analyzed. Although all adolescent males were unequivocally subordinate to all adult males, dominance relationships within the age classes were much less clear. Especially among adolescent males, few pant-grunts or agonistic interactions occurred. While adolescent males frequently pant-grunted at adult males, these latter males, except the alpha and the youngest, rarely pant-grunted to one another. This suggests that a difference of social status exists between adolescent and adult males. Adult males rarely display overt dominance to one another probably because the presence of other males affects their interactions. Moreover, they seem to try to keep their dominance relationship ambiguous when making it overt is not advantageous to them. This may be a political way for males to coexist with one another in a unit-group.     

Glycogen phosphorylase ‘b’ in Dictyostelium: stability and endogenous phosphorylation

Summary The slime mold Dictyostelium discoideum has two forms of the enzyme glycogen phosphorylase. The inactive phosphorylase b form requires 5 AMP for activity and is present in early development. The active phosphorylase a form is 5 AMP independent and occurs during later development. We here show that the 92 kd b enzyme subunit exists either as a singlet or a doublet upon SDS-PAGE, depending on the method of sample extraction. In the presence of exogenously added Mn²⁺ and ATP, the phosphorylase b shows apparent conversion into a 5 AMP independent form as measured by enzyme activity. In addition, Mn²⁺ and ATP also support an in vitro phosphorylation of the 92 kd phosphorylase b subunit. We also demonstrate phosphorylation of the b enzyme subunit in vivo by ³²-P incorporation into the enzyme protein. A protein kinase responsible for the observed in vitro phosphorylation of the phosphorylase b subunit is characterized.     

Hemolytic and coagulation properties of Sphaeridiotrema globulus (Trematoda)

Metacercariae of Sphaeridiotrema globulus (Trematoda) were obtained from naturally infected Goniobasis virginica (Pleuroceridae). Excysted metacercariae were placed individually on blood-agar plates. After 8 hr of incubation at 42 C the plates exhibited beta-hemolysis. The reaction zone was approximately 1.5 mm in diameter. Laboratory-reared mallard ducks (Anas platyrhynchos) were infected with S. globulus. Mallards developed fatal sphaeridiotremiasis and demonstrated increased prothrombin time on days 3 and 6 postinfection.     

The effects of crowding on adults of Echinostoma revolutum (Digenea: Echinostomatidae) in experimentally infected golden hamsters, Mesocricetus auratus

All 30 female golden hamsters, Mesocricetus auratus, fed either 125 +/- 50 (group A), 300 +/- 50 (group B), or 500 +/- 50 (group C) metacercarial cysts of Echinostoma revolutum were infected 7-35 days postexposure. The mean number of worms in A, B, and C were 62, 96, and 212, respectively. Most of the worms in A were in the jejunum, but in C worms were about equally distributed in the duodenum, jejunum, and ileum, and some were in the cecum. The body area and wet and dry weights of worms from C were significantly less than that of A or B at 2, 4, and 5 wk postinfection. Echinostoma revolutum eggs were in the feces of 100% of the hamsters by days 12, 13, and 14 in A, B, and C, respectively.