Nodal cis-regulatory elements reveal epiblast and primitive endoderm heterogeneity in the peri-implantation mouse embryo

Nodal, a secreted factor known for its conserved functions in cell-fate specification and the establishment of embryonic axes, is also required in mammals to maintain the pluripotency of the epiblast, the tissue that gives rise to all fetal lineages. Although Nodal is expressed as early as E3.5 in the mouse embryo, its regulation and functions at pre- and peri-implantation stages are currently unknown. Sensitive reporter transgenes for two Nodal cis-regulatory regions, the PEE and the ASE, exhibit specific expression profiles before implantation. Mutant and inhibitor studies find them respectively regulated by Wnt/β-catenin signaling and Activin/Nodal signaling, and provide evidence for localized and heterogeneous activities of these pathways in the inner cell mass, the epiblast and the primitive endoderm. These studies also show that Nodal and its prime effector, FoxH1, are not essential to preimplantation Activin/Nodal signaling. Finally, a strong upregulation of the ASE reporter in implanting blastocysts correlates with a downregulation of the pluripotency factor Nanog in the maturing epiblast. This study uncovers conservation in the mouse blastocyst of Wnt/β-catenin and Activin/Nodal-dependent activities known to govern Nodal expression and the establishment of polarity in the blastula of other deuterostomes. Our results indicate that these pathways act early on to initiate distinct cell-specification processes in the ICM derivatives. Our data also suggest that the activity of the Activin/Nodal pathway is dampened by interactions with the molecular machinery of pluripotency until just before implantation, possibly delaying cell-fate decisions in the mouse embryo.     

Allocation of inner cells to epiblast vs primitive endoderm in the mouse embryo is biased but not determined by the round of asymmetric divisions (8→16- and 16→32-cells)

The epiblast (EPI) and the primitive endoderm (PE), which constitute foundations for the future embryo body and yolk sac, build respectively deep and surface layers of the inner cell mass (ICM) of the blastocyst. Before reaching their target localization within the ICM, the PE and EPI precursor cells, which display distinct lineage-specific markers, are intermingled randomly. Since the ICM cells are produced in two successive rounds of asymmetric divisions at the 8→16 (primary inner cells) and 16→32 cell stage (secondary inner cells) it has been suggested that the fate of inner cells (decision to become EPI or PE) may depend on the time of their origin. Our method of dual labeling of embryos allowed us to distinguish between primary and secondary inner cells contributing ultimately to ICM. Our results show that the presence of two generations of inner cells in the 32-cell stage embryo is the source of heterogeneity within the ICM. We found some bias concerning the level of Fgf4 and Fgfr2 expression between primary and secondary inner cells, resulting from the distinct number of cells expressing these genes. Analysis of experimental aggregates constructed using different ratios of inner cells surrounded by outer cells revealed that the fate of cells does not depend exclusively on the timing of their generation, but also on the number of cells generated in each wave of asymmetric division. Taking together, the observed regulatory mechanism adjusting the proportion of outer to inner cells within the embryo may be mediated by FGF signaling.     

Early gonadal development and primary males in the protogynous epinepheline, Cephalopholis boenak

Early gonadal development of the protogynous epinepheline, Cephalopholis boenak, was examined histologically in 289 specimens with standard length (L_S) of 42–130 mm, collected from May 2000 to April 2002 in Hong Kong waters, to determine male developmental pathways and establish its sexual pattern. All juvenile gonads developed an ovarian lumen with primary‐growth stage oocytes and scattered spermatogenic tissue prior to sexual differentiation and first sexual maturation. From this bisexual phase containing both female and male tissues, some gonads differentiated as ovaries with further oocyte growth to cortical‐alveolus and vitellogenic stages, the rest differentiated as testes with the proliferation of spermatogenic tissue and the formation of a sperm sinus. All testes retained the lumen and primary‐growth stage oocytes, and sperm sinuses ran within the gonad wall. Unlike most protogynous species, among functional males it was impossible to distinguish those resulting from juveniles through sexual differentiation (i.e. primary male) from those resulting from functional females through sex change (i.e. secondary male) based solely on testicular morphology. A proportion‐spermatogenic‐tissue index (I_ST) was, therefore, developed and determined to be a reliable quantitative indicator for distinguishing differentiating, primary males before a sperm sinus was evident, from differentiating females during sexual differentiation. Since sexually transitional specimens with the concominant appearance of degenerating vitellogenic, or later, stage oocytes and spermatogenic tissue in the gonads were previously noted from Hong Kong, diandric, protogynous hermaphroditism is confirmed in C. boenak. For species, such as this and other epinephelines, in which all males have the same testicular morphology, a complete analysis of a wide range of body sizes from juveniles to adults is necessary for understanding male developmental pathways, and determining sexual pattern.     

Sex-Related Differences in Blood and Gonad Levels of Testosterone in Silver Fox Fetuses

The mass of silver fox fetuses of both sexes, their gonads, and adrenals, and the levels of testosterone in the blood serum and in gonads and adrenals were determined from day 31 of gestation and every five days thereafter until its termination. Marked sex-related differences were revealed: the blood and gonad levels of testosterone in male fetuses were much higher than those in female fetuses. The fetal adrenals contained significantly less testosterone than the gonads. No sex-related differences in the content of testosterone in the fetal adrenals were found. No differences were found in the body and adrenal mass in female and male fetuses at all the developmental stages studied, while the mass of ovaries exceeded that of testes from day 45 of gestation. The data obtained suggest sex dimorphism in the production of testosterone by gonads in silver foxes appears after day 35 and appears to correspond to the period of morphological differentiation of gonads.     

Histological study and immunohistochemical location of cytoskeletal proteins in the testis and epididymis of the three species of lizards of the family Leiosauridae (Reptilia: Squamata)

The objective of the study was to histologically describe the structure of the testis and epididymis, identifying the GAGs in them. Furthermore, the distribution of smooth muscle alpha-actin, desmin, and vimentin is described of E. bilineatus, E. perditus, and U. vautieri, through immunohistochemical method. Two adult males of each species of the lizards were collected in southeastern Brazil. In the testicular albuginea, the presence of neutral glycoproteins was detected. In E. bilineatus, the epididymal duct epithelium is simple, composed of secretory cells, cylindrical in shape and acidophilic cytoplasm. In the testis of the three lizards there was a positive immunolocation accentuated for α-SMA in the dense connective tissue of the tunica albuginea. A moderate reaction was observed in the connective tissue albuginous and in the layer of smooth muscle fibers in a circumferential arrangement of the epididymal duct. The immunolocation for desmin was accentuated in the fibers of the testicular albuginea of E. bilineatus, while in U. vautier and E. perditus was moderate. In the epididymis, only in E. perditus and E. bilineatus positive immunoreaction was verified. In the three lizards there were no observations of immunostaining for vimentin with the antibody utilized.     

原始生物大分子动态的数学模型及其进化意义

本文应用生态学上关于种群增长及种间竞争的逻辑斯缔方程,提出原始生物大分子动态的数学模型,对从该模型推导得的几种可能结果进行了分析,得出在生命起源初期的原始生物圈的生物多样性是很低的结论,即原始生物圈中较大量存在的只是一些种类单一的,可复制的原始生物大分子,因此,可以认为生物进化就是建立在这样的一个基础上,并沿着生物多样性不断提高的途径演化,该模型还有助于对广泛存在于现代生物基因组中的重复序列的起源的认识。     

Origin of life between scylla and charybdis

Summary The package model discussed here is concerned with the preservation of genetic information by primordial compartments. Each viable package encloses a complete set of unlinked genes in varying numbers of copies. Due to stochastic distribution and error-prone replication two potent perils endanger the informational integrity of packages: fluctuation and mutation.A computer simulation was used to quantify the effects of fluctuation, mutation, and package death by accident. Assuming reasonable rates for these parameters it is suggested that life started out with compartments containing not more than 3 different genes.Abbreviations A adenine - C cytosine - G guanine - U uracil - N any one of these 4 bases     

Polymerization of amino acids under primitive earth conditions

Summary Chemical evolution on the primitive earth must have involved the condensation of-amino acids to peptides under a variety of conditions. Subjecting a mixture of methane, ammonia, and water to an electric discharge in the presence of free amino acids yields small peptides. The dehydration-condensation may have taken place via ammonium cyanide, the hydrogen cyanide tetramer, or aminonitriles. The experiments may be considered genuinely prebiotic and significant in the context of chemical evolution.