Bivariate flow karyotyping of human chromosomes: evaluation of variation in Hoechst 33258 fluorescence, chromomycin A3 fluorescence, and relative chromosomal DNA content.

The total variation of chromosome peak positions, in bivariate distributions of Hoechst 33258 and chromomycin A3 fluorescence of 19 healthy individuals, was compared with the experimental variation, determined from 23 bivariate distributions of chromosomes prepared separately from a single cell lineage. The experimental variation in Hoechst and chromomycin fluorescence and the relative chromosomal DNA content were determined from experiments performed over several days. The additional variance contributed by time was the same as the daily variance. The accuracy by which the relative chromosomal DNA content can be calculated from bivariate peak positions was investigated. A least squares method was used to fit the distributions of relative DNA content, obtained, respectively, from mono- and bivariate flow analyses of chromosomes from the same cell lineage. In general the DNA contents match quite well, but for a few chromosomes a difference was found, statistically discernible at the 5% level. The average relative chromosomal DNA content of the chromosomes from the 19 normal individuals, calculated from bivariate peak positions, showed a linear relation with the estimates published by other investigators.     

Structure of the bovine eye lens gamma s-crystallin gene (formerly beta s)

The organization of a number of crystallin genes has already been resolved. One of the remaining genes of which the structure was hitherto unknown is the gamma s gene (formerly beta s). We determined the complete sequence of the bovine gamma s-crystallin-coding gene, apart from the middle region of the first intron. Since it contains three exons and two introns, we conclude that the former beta s, also at the gene level is gamma-crystallin-like. However, it is located on chromosome 3, in contrast to other gamma genes which occur in tandem on the human chromosome 2.     

Presence of hybridizing DNA sequences homologous to bovine acidic and basicβ-crystallins in all classes of vertebrates

Summary The eye lens-crystallins in cow and chicken are encoded by a family of at least six genes. In order to assess the distribution of the corresponding genes among other vertebrates we hybridized -crystallin sequences (A2, A3/A1, A4, B1, B2, B3), isolated from a bovine lens cDNA library, to Southern blots on whichEcoR1-digested chromosomal DNA was blotted from different vertebrate species. These included human, chimpanzee, calf, rat, pigeon, duck, monitor lizard, toad, trout, and lamprey. Positive hybridization signals were found in the representatives of virtually all classes of vertebrates. The basic B-crystallins gave hybridization signals in more species than the acidic A ones. In monitor lizard and toad the weakest hybridization signals for basic crystallin probes were found. For acidic crystallin probes the distribution pattern was more simple; among cold-blooded vertebrates a signal for A2 was found in trout and lamprey, for A4 in trout, and for A3/A1 only in toad. The results demonstrate that the duplications leading to the -crystallin gene family occurred before or during the earliest stages of vertebrate evolution.     

Piglet and placental traits at term in relation to the estrogen receptor genotype in gilts

Liveborn piglets from gilts with estrogen receptor (ESR) genotype AA (95 AA-AA and 91 AA-AB piglets), AB (88 AB-AA, 118 AB-AB, and 37 AB-BB piglets), and BB (97 BB-AB and 89 BB-BB piglets) were compared after farrowing, to examine whether piglet ESR genotype (ESRp) nested within maternal ESR genotype (ESRm) affected placental traits at term, piglet birth weight, and growth until weaning. Furthermore, the relation of birth weight to various placental traits and the relations between placental traits were evaluated relative to ESR genotype. For this study, 62 Large White x Meishan F2 crossbred gilts (18 AA, 24 AB, and 20 BB) were used. The gilts belonged to a population in which the A allele is favorable for litter size. ESRp nested within ESRm did not affect placental length, weight, surface area and number of areolae. ESRp nested within ESRm affected amnion weight (AA-AA amnions were heavier than AA-AB, AB-AA and BB-AB amnions), placental weight after including placental surface area in the model (AA-AB placentae were lighter than AA-AA, AB-BB and BB-AB placentae), placental efficiency calculated as birth weight divided by placental weight (AB-AA placentae were less efficient than AA-AB placentae), and the relations of birth weight to placental weight and birth weight to number of areolae. The found differences imply an interaction of maternal and fetal ESR genotype on placental traits (especially weight and number of areolae) during fetal development. Furthermore, the found effects on placental and amnion weight might be the result of a difference in thickness or vascularization or both. The favorable ESR allele for litter size, i.e. the A allele, appears to be the unfavorable allele for pre-weaning piglet growth. Therefore, further research on ESR in relation to vascularization, weight and thickness of placentae. uterine size, endometrial gland development, and piglet growth is recommended.     

The monotreme genome: a patchwork of reptile, mammal and unique features?

The first specimen of platypus (Ornithorhynchus anatinus) that reached Britain in the late 18th century was regarded a scientific hoax. Over decades the anatomical characteristics of these unique mammals, such as egg laying and the existence of mammary glands, were hotly debated before they were accepted. Within the last 40 years, more and more details of monotreme physiology, histology, reproduction and genetics have been revealed. Some show similarities with birds or reptiles, some with therian mammals, but many are very specific to monotremes. The genome is no exception to monotreme uniqueness. An early opinion was that the karyotype, composed of a few large chromosomes and many small ones, resembled bird and reptile macro- and micro-chromosomes. However, the platypus genome also features characteristics that are not present in other mammals, such as a complex translocation system. The sex chromosome system is still not resolved. Nothing is known about dosage compensation and, unlike in therian mammals, there seems to be no genomic imprinting. In this article we will recount the mysteries of the monotreme genome and describe how we are using recently developed technology to identify chromosomes in mitosis, meiosis and sperm, to map genes to chromosomes, to unravel the sex chromosome system and the translocation chain and investigate X inactivation and genomic imprinting in monotremes.     

Fetal and placental traits at day 35 of pregnancy in relation to the estrogen receptor genotype in pigs

Fetuses from gilts with estrogen receptor (ESR) genotype AA (AA-AA and AA-AB) and BB (BB-AB and BB-BB) were compared at Day 35/36 of pregnancy, to examine whether fetal ESR genotype nested within maternal ESR genotype would affect fetal traits. Furthermore the relation of fetal body weight and fetal heart weight to various placental traits were evaluated relative to ESR genotype. Fetal and placental weight and length, and implantation surface area were not affected by fetal ESR genotype nested within maternal ESR genotype. Fetal weight was related similarly to placental length, placental weight, and implantation surface area: up to a certain threshold value (40 cm, 40 g and 250 cm2, respectively), an increase in the trait was associated with an increase of fetal weight. Thereafter, fetal weight did not change anymore. Thus, at Day 35/36 of pregnancy porcine fetuses seem to have a maximum growth potential. The percentage of AA-AA fetuses that had not reached this maximum growth potential was larger than of the other three genotype combinations studied, and therefore a higher subsequent fetal mortality may be expected in this group. Hearts of AA-AB fetuses were significantly heavier than those of BB-AB and BB-BB fetuses and tended to be heavier than those of AA-AA fetuses. The reason for this hypertrophy is unclear, but might be related to a difference in placental vascularity. Heart weight of fetuses from BB gilts increased with fetal weight, while heart weights of fetuses from AA gilts did not. Heart weight increased with an increase of placental length and implantation surface area up to 51 cm and 437 cm2, respectively, and thereafter decreased again. For BB-AB fetuses a similar relation was found between heart weight and placental weight, while heart weight of the other three genotype combinations remained unaffected as placental weight increased. The fetus and placenta are continuously changing during early pregnancy, therefore different mechanisms may change the demands for cardiac output. However, keeping in mind that placental size and blood volume are relatively large, placental vascularity and vascular development may play a major role. Therefore, further research on heart size, placental size and vascularity, relative to ESR genotype, is recommended.     

The evolution of imprinting: chromosomal mapping of orthologues of mammalian imprinted domains in monotreme and marsupial mammals

Background  

The evolution of genomic imprinting, the parental-origin specific expression of genes, is the subject of much debate. There are several theories to account for how the mechanism evolved including the hypothesis that it was driven by the evolution of X-inactivation, or that it arose from an ancestrally imprinted chromosome.     

Cytokine production by leukocytes of military personnel with depressive symptoms after deployment to a combat-zone: a prospective, longitudinal study

Major depressive disorder (MDD) is frequently diagnosed in military personnel returning from deployment. Literature suggests that MDD is associated with a pro-inflammatory state. To the best of our knowledge, no prospective, longitudinal studies on the association between development of depressive symptomatology and cytokine production by peripheral blood leukocytes have been published. The aim of this study was to investigate whether the presence of depressive symptomatology six months after military deployment is associated with the capacity to produce cytokines, as assessed before and after deployment. 1023 military personnel were included before deployment. Depressive symptoms and LPS- and T-cell mitogen-induced production of 16 cytokines and chemokines in whole blood cultures were measured before (T0), 1 (T1), and 6 (T2) months after return from deployment. Exploratory structural equation modeling (ESEM) was used for data reduction into cytokine patterns. Multiple group latent growth modeling was used to investigate differences in the longitudinal course of cytokine production between individuals with (n = 68) and without (n = 665) depressive symptoms at T2. Individuals with depressive symptoms after deployment showed higher T-cell cytokine production before deployment. Moreover, pre-deployment T-cell cytokine production significantly predicted the presence of depressive symptomatology 6 months after return. There was an increase in T-cell cytokine production over time, but this increase was significantly smaller in individuals developing depressive symptoms. T-cell chemokine and LPS-induced innate cytokine production decreased over time and were not associated with depressive symptoms. These results indicate that increased T-cell mitogen-induced cytokine production before deployment may be a vulnerability factor for development of depressive symptomatology in response to deployment to a combat-zone. In addition, deployment to a combat-zone affects the capacity of T-cells and monocytes to produce cytokines and chemokines until at least 6 months after return.