Alteration of SREBP activation in liver of trisomy 21 fetuses.

We previously reported that trisomy 21 (T21) fetuses have an intrinsic lipid metabolism abnormality resulting in higher serum cholesterol levels than their matched controls. In an attempt to clarify the biochemical basis of this derangement we analyzed the liver cholesterol levels and activation of the sterol regulatory element binding proteins SREBP-1 and SREBP-2. We report here for the first time that SREBP-1 and SREBP-2 are present in human fetal liver and their activation follows a different regulatory pattern. Moreover T21 fetuses show a peculiar pattern of SREBP activation which, at variance from control fetuses, involves sterol-independent maturation of SREBP-1. Multiple defects accompanied the lipid derangement in T21, resulting in high circulating and tissue cholesterol. This may serve as an early biochemical marker of an unknown, possibly genetically determined mechanism, whose consequence on lipid homeostasis during postnatal and adult life is still not understood.     

Distribution of extracellular matrix components in nuchal skin from fetuses carrying trisomy 18 and trisomy 21

We have investigated histologically the elevations of the skin in dorsal and lateral neck (nuchal) regions of human fetuses carrying karyotypes of trisomy 18 (Edwards' syndrome) and trisomy 21 (Down's syndrome). Cavities filled with interstitial fluid were found in the dermis, epidermal basement membrane and occasionally in the epidermis of trisomy-18 fetuses, but were not delineated by an epithelium or basement membrane as judged by the absence of immunostaining for laminin, collagen IV and collagen VII. Dilated vessels were also found at the interface between dermis and subcutis. Neither normal fetal skin nor that of trisomy-21 fetuses contained cavities or dilated vessels. In order to detect possible alterations of the extracellular matrix in trisomy-18 and trisomy-21 skin, the distribution of glycoproteins, glycosaminoglycans and proteoglycans was studied immunohistochemically. In trisomy-21 and control skin, the dermis stained intensely for fibronectin, whereas the subcutis reacted only weakly. In trisomy-18 skin, the stronger staining for fibronectin appeared in the subcutis, and the prevailing collagen type was collagen III, collagen type I being absent. In the skin of trisomy-21 fetuses, collagen VI was more irregularly arranged and densely packed, whereas collagen I was more widely spaced than in normal fetuses. More hyaluronan was present in the dermis and subcutis of trisomy-21 fetuses than in that of trisomy-18 and control fetuses. A correlation seems to exist between undelimited cavities and collagen III in trisomy-18 skin, and between hyaluronan and the specific arrangement of collagen in trisomy-21 skin.Abbreviations bm Basement membrane - ep epidermis - d dermis - sc subcutis - hf hair follicle - c capillary This article is dedicated to Professor Dr. Konrad Märkel on the occasion of his 70^th birthday     

Partial trisomy 21

Summary Cytogenetic analysis of a 6-year-old girl with moderate mental retardation revealed 46 chromosomes with a tandem translocation (21;21) resulting in a partial trisomy 21. Only the terminal band 21q22 was not in triplicate. G-, Q-, R-, and C-banding techniques and silver nitrate staining of the nucleolus organizer regions (NORs) were used to identify this chromosome fully.The phenotype of the patient was not typical for Down's syndrome, providing additional evidence that trisomy of band 21q22 is pathogenetic for the phenotype of Down's syndrome. This is also a new example in human pathology of a stable dicentric chromosome in which one of the centromeric constrictions appears to be nonfunctional.     

Altered DNA methylation in leukocytes with trisomy 21

The primary abnormality in Down syndrome (DS), trisomy 21, is well known; but how this chromosomal gain produces the complex DS phenotype, including immune system defects, is not well understood. We profiled DNA methylation in total peripheral blood leukocytes (PBL) and T-lymphocytes from adults with DS and normal controls and found gene-specific abnormalities of CpG methylation in DS, with many of the differentially methylated genes having known or predicted roles in lymphocyte development and function. Validation of the microarray data by bisulfite sequencing and methylation-sensitive Pyrosequencing (MS-Pyroseq) confirmed strong differences in methylation (p<0.0001) for each of 8 genes tested: TMEM131, TCF7, CD3Z/CD247, SH3BP2, EIF4E, PLD6, SUMO3, and CPT1B, in DS versus control PBL. In addition, we validated differential methylation of NOD2/CARD15 by bisulfite sequencing in DS versus control T-cells. The differentially methylated genes were found on various autosomes, with no enrichment on chromosome 21. Differences in methylation were generally stable in a given individual, remained significant after adjusting for age, and were not due to altered cell counts. Some but not all of the differentially methylated genes showed different mean mRNA expression in DS versus control PBL; and the altered expression of 5 of these genes, TMEM131, TCF7, CD3Z, NOD2, and NPDC1, was recapitulated by exposing normal lymphocytes to the demethylating drug 5-aza-2'deoxycytidine (5aza-dC) plus mitogens. We conclude that altered gene-specific DNA methylation is a recurrent and functionally relevant downstream response to trisomy 21 in human cells.     

Gonadal function in trisomy 21

Endocrinologic evaluation of 39 patients with trisomy 2 and associated hypogonadism demonstrated elevations of follicle-stimulating hormone and luteinizing hormone; consequently, it can be concluded that both germinal and Leydig cell function are affected. A negative correlation between testicular size and plasma follicle-stimulating hormone was documented. Plasma testosterone levels were found to be normal in male patients as were estradiol levels in female patients with trisomy 21. On the basis of these findings, the simplest and most practical diagnostic approach to evaluate germinal cell function appears to be a single plasma follicle-stimulating hormone determination supplemented by an accurate measurement of testicular volume in males.     

Recurrences of trisomy 18 and trisomy 13 after trisomy 21

Summary Between 40 years and 43 years of age, a woman had three consecutive pregnancies with different prenatally diagnosed autosomal trisomies. This is compatible with the view that the predisposition to non-disjunction is not chromosome-specific.     

Telomeres in trisomy 21 amniocytes

Individuals with trisomy 21 have an increased risk of developing leukemia and premature dementia. They also have a higher rate of telomere loss. The aim of the study was to compare telomere length and the hTERC gene copy number, which encodes the telomerase RNA subunit, in amniocytes of trisomy 21 conceptions and normal pregnancies. A quantitative fluorescence-in-situ protocol (Q-FISH) was used to compare telomere length in amniocytes cultured from 11 trisomy 21 conceptions and from 14 normal pregnancies. Quantification was conducted using novel computer software. Fluorescence in situ hybridization (FISH) was used to assess the percentage of cells with additional copies of hTERC. We found that the immunofluorescence intensity, which represents telomere length, was significantly lower in amniocytes from trisomy 21 conceptions compared to the control group. The trisomy 21 group had a higher number of cells with additional copies of hTERC. This observation could be one of the cytogenetic parameters that represent a state of genetic instability and might play a role in the pathomechanism of typical features of Down syndrome, such as dementia and malignancy.     

Anomalies in thyroid function in children with trisomy 21

Thyroid function of 60 children with Down (DS) aged 3 months to 16 years was studied by evaluation of serum concentration of ultra-sensitive thyroid stimulating hormone (TSH), free T4 and T3 (FT4, FT3), total T4 and T3 (T4 and T3) and reverse T3 (rT3). Each DS child was matched to a control of the same age. The concentration of TSH was increased in DS children while the concentration of rT3 of the DS children was significantly decreased compared to the controls as was the ratio rT3/TSH. These results showed that thyroid function of DS children is abnormal.     

Alpha-fetoprotein and trisomy 21

In 31 affected pregnancies with Down syndrome, the median maternal serum alpha-fetoprotein value was lower than normal, 0.76 MoM, and median amniotic fluid value was quite normal, 0.98 MoM. Selecting an arbitrary cutoff-point of 0.5 MoM, 4.1 percent of normal gestations show values less than 0.5 MoM. Authors discuss problems about screening for fetal Down's syndrome by measuring maternal serum AFP levels.     

Phospholipid methylation in fibroblasts of patients with trisomy 21

Comparative studies of phospholipid methylation in normal and Down syndrome (trisomy 21) fibroblasts were performed on a series of 7 pairs. Two patients showed an increased phospholipid methylation pathway as compared to the appropriate controls; one patient showed a decreased pathway; and in four patients there was no significant difference. The phospholipid methylation pathway appears to be a minor pathway, as compared to the choline pathway of phosphatidylcholine synthesis and thus does not allow the detection of an eventual disturbance of monocarbon metabolism in Down syndrome.     

Isochromosome not translocation in trisomy 21q21q

Summary After primary trisomy, de novo 21q21q trisomy is the most frequent chromosomal aberration responsible for Down syndrome. This rearrangement is more commonly referred to as a Robertsonian translocation or centric fusion product than as an isochromosome, e.g., t(21q;21q) instead of i(21q); however, in practice, it has not so far proved possible to distinguish between these alternatives. The aim of this work was to establish which of the two alternatives is acceptable.     

Altered calcium currents in cultured sensory neurons of normal and trisomy 16 mouse fetuses, an animal model for human trisomy 21 (Down syndrome)

Down syndrome is determined by the presence of an extra copy of autosome 21 and is expressed by multiple abnormalities, with mental retardation being the most striking feature. The condition results in altered electrical membrane properties of fetal dorsal root ganglia (DRG) neurons, as in the trisomy 16 fetal mouse, an animal model of the human condition. Cultured trisomic DRG neurons from human and mouse fetuses present faster rates of depolarization and repolarization in the action potential compared to normal controls and a shorter spike duration. Also, trisomy 16 brain and spinal cord tissue exhibit reduced acetylcholine secretion. Therefore, we decided to study Ca2+ currents in cultured DRG neurons from trisomy 16 and age-matched control mice, using the whole-cell patch-clamp technique. Trisomic neurons exhibited a 62% reduction in Ca2+ current amplitude and reduced voltage dependence of current activation at -30 and -20 mV levels. Also, trisomic neurons showed slower activation kinetics for Ca2+ currents, with up to 80% increase in time constant values. Kinetics of the inactivation phase were similar in both conditions. The results indicate that murine trisomy 16 alter Ca2+ currents, which may contribute to impaired cell function, including neurotransmitter release. These abnormalities also may alter neural development.     

Parental trisomy 21 mosaicism.

A family with three children with trisomy 21 in which the mother is a phenotypically normal, trisomy 21/normal mosaic was studied. Chromosome 21 fluorescent heteromorphisms were used to document that two of the three number 21's in two of the Down syndrome offspring were of maternal origin. Five cytogenetic surveys in which both parents of a child with trisomy 21 were studied have been reviewed. From these data, it is estimated that 3% of couples producing a child with trisomy 21 can be explained by parental mosaicism. From 17 informative sibships, with one parent mosaic, the segregation ratio was estimated to be 0.43 +/- 0.11.     

Reciprocal translocations and full trisomy (trisomy 18 and trisomy 21) in the offspring

In this report, we present examples of trisomy 18 and trisomy 21, both resulting from maternal reciprocal translocations: 46, XX, t(5;18) (q21;q11) and 46, XX, t(5;21) (p11.2;p11), respectively.     

Partial trisomy 21 (21q21 - 21q22.2)]

An abnormal chromosome 21 is reported in a child with a phenotype strongly reminiscent of trisomy 21 syndrome. It is shown to result from duplication of the segment 21q21 leads to 21q22.2. Comparison of the phenotype with that of other partial and total trisomics shows that the characteristic features of the trisomy 21 syndrome (mongolism), the mental retardation in particular - is due to trisomy 21q22.2 and perhaps 21q22.2.     

Partial trisomy and monosomy 21 in an infant with an unusual de novo 21/21 translocation

A 3-month-old boy with a 46,XY,--21,+t(21;21)(pter leads to q22.3::q22.3 leads to q11::p11 leads to pter) karyotype, implicating trisomy for the 21q11 leads to 21q22.2 segment and monosomy for the 21q22.3 sub-band, is described. Most of the clinical features corresponded to Down syndrome ; other signs such as large ears, prominent nasal bridge and retromicrognathia were interpreted as the expression of 21q22.3 monosomy. The abnormal monocentric chromosome had satellites and stalks on both ends as a result of a 21q;21q translocation followed by deletion of one centromere region. Despite similar stalk size and NOR-Ag positiveness a significantly higher association frequency of the centrometric end as compared to the acentric end was found. This observation suggests that the satellite association phenomenon is not exclusively NOR-dependent, but that the centromeric and/or p11 regions of acrocentrics also play an important role.     

Meiotic configurations in female trisomy 21 foetuses

Summary Analysis of the meiotic configurations formed by the three No 21 chromosomes in oocytes from two trisomy 21 foetuses was undertaken using a spreading technique. Light microscope analysis of the first gave limited resolving power, such that over half the oocytes could not be classified as to presence or absence of trivalent or bivalent plusunivalent. In the second, investigated at the electron microscope level, all 65 cells analysed were informative and precise detail of meiotic pairing in trivalents could be obtained. Two principal forms of trivalent occurred, one in which pairing was initiated at opposite ends of the three No 21's, each initiation point involving only two of the three homologous lateral elements; the other in which pairing was initiated by all three elements at the same end, a triple synaptonemal complex being formed. Only in one oocyte out of the 65 analysed at EM level, however, did triple pairing occur along the entire length of the No 21 trivalent. All others showed splitting into bivalent and univalent at some point along the structure. Unpaired regions within trivalents and all univalents were consistently seen to be thickened and dark staining with silver over the whole period from pachytene to diplotene. This contrasted with the desynapsing lateral elements of previously paired synaptonemal complexes which appeared thin by comparison at diplotene. The significance of the thickening remains, as yet, obscure.