Microdissection and Dop-PCR-Based Reverse Chromosome Painting as a Fast and Reliable Strategy in the Analysis of Various Structural Chromosome Abnormalities

Müller‐Navia, Jutta; Nebel, Almut; Oehler, Detlef; Theile, Ursel; Zabel, Bernhard; Schleiermacher, E.

doi:10.1002/(sici)1097-0223(199610)16:10<915::aid-pd966>3.0.co;2-v

Cited by 21 publications

(7 citation statements)

References 32 publications

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“…The majority of WHS-associated chromosome changes are de novo, and they are expected to be isolated deletions. However, unbalanced de novo translocations, consisting more often of t(4p;8p) translocations, but also including a t(4p;7p) translocation, have been reported with unexpected high frequency in WHS, and they were all maternal in origin (Wieczorek et al 2000;Zollino et al 2004;Tonnies et al 2001;Petit et al 1998;Muller-Navia et al 1996). These reports point at a great complexity of the WHS-associated chromosome defects, which can be the cause of the great phenotypic variability of this condition.…”

Section: Discussionmentioning

confidence: 99%

Wolf–Hirschhorn syndrome-associated chromosome changes are not mediated by olfactory receptor gene clusters nor by inversion polymorphism on 4p16

et al. 2007

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The basic genomic defect in Wolf-Hirschhorn syndrome (WHS), including isolated 4p deletions and various unbalanced de novo 4p;autosomal translocations and above all t(4p;8p), is heterogeneous. Olfactory receptor gene clusters (ORs) on 4p were demonstrated to mediate a group of WHS-associated t(4p;8p)dn translocations. The breakpoint of a 4-Mb isolated deletion was also recently reported to fall within the most distal OR. However, it is still unknown whether ORs mediate all 4p-autosomal translocations, or whether they are involved in the origin of isolated 4p deletions. Another unanswered question is whether a parental inversion polymorphism on 4p16 can act as predisposing factor in the origin of WHS-associated rearrangements. We investigated the involvement of the ORs in the origin of 73 WHS-associated rearrangements. No hotspots for rearrangements were detected. Breakpoints on 4p occurred within the proximal or the distal olfactory receptor gene cluster in 8 of 73 rearrangements (11%). These were five t(4p;8p) translocations, one t(4p;7p) translocation and two isolated terminal deletions. ORs were not involved in one additional t(4p;8p) translocation, in a total of nine different 4p;autosomal translocations and in the majority of isolated deletions. The presence of a parental inversion polymorphism on 4p was investigated in 30 families in which the 4p rearrangements, all de novo, were tested for parental origin (7 were maternal and 23 paternal). It was detected only in the mothers of 3 t(4p;8p) cases. We conclude that WHS-associated chromosome changes are not usually mediated by low copy repeats. The 4p16.3 inversion polymorphism is not a risk factor for their origin.

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Section: Discussionmentioning

confidence: 99%

Wolf–Hirschhorn syndrome-associated chromosome changes are not mediated by olfactory receptor gene clusters nor by inversion polymorphism on 4p16

et al. 2007

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“…(ii) For reverse painting five copies of the derivative chromosome 14 (derived from peripheral blood lymphocyte metaphases of the propositus) were microdissected and amplified by DOP‐PCR as described by Guan et al [1994]. Amplified microdissected DNA was labeled with Biotin‐16‐dUTP (Roche Diagnostics) in a secondary PCR reaction according to the protocol of Müller‐Navia et al [1996]. Chromosomal in situ suppression (CISS) hybridization and detection of the biotinylated probes was carried out as described [Lichter and Cremer, 1992] with minor modifications.…”

Section: Methodsmentioning

confidence: 99%

Mental and psychomotoric retardation in two brothers with pure partial trisomy 7q32-q34 due to a maternal insertion (14;7)

et al. 2000

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We present two brothers with mental retardation, seizures disorder, generalized muscular hypertonia, kyphoscoliosis, minor anomalies and a prominent midface. GTG-banded chromosome analysis showed a derivative chromosome 14 without clues toward the origin of the rearrangement. Microdissection of the derivative chromosome 14 and subsequent reverse painting demonstrated partial trisomy 7q32-q34 as the unbalanced product of a maternal insertion (14;7). Thus, we identified two cases with pure trisomy 7q32-q34 that allowed further delineation of this aneusomy syndrome.

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“…Theoretically, a single cell contains ample nuclear material for PCR amplification. Whilst a number of methods for single cell PCR have been described, 13,[25][26][27][28] the small quantity of template obtained from such specimens is not readily responsive to standard methods of manipulation. Until reliable and reproducible methods for such specimens are more common-place, molecular analysis of microdissected tissue requires that many cells be acquired for each experiment.…”

Section: Downstream Applications For Microdissected Specimensmentioning

confidence: 99%

New developments in the analysis of gene expression

Burgess

Hazelton

2000

Redox Report

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An understanding of the relationship between gene expression, protein expression and the influences of genetic responses upon gene function is vital before we can understand the complexity of genomes. Traditional methods for the study of gene expression are limited to studying small groups of genes at a time and a source of pure starting material has been difficult to obtain. Recent technological advances have enabled large numbers of genes, from specific cell populations, to be studied in a single experiment. Laser capture microdissection (LCM) and microarray technology are providing the next revolution in the study of gene expression. LCM-based molecular analysis of histopathological lesions can be applied to any disease process that is accessible through tissue sampling. Examples include: (i) mapping the field of genetic changes associated with oxidative stress; (ii) analysis of gene expression patterns in atherosclerotic tissues, sites of inflammation and Alzheimer's disease plaques; (iii) infectious micro-organism diagnosis; and (iv) typing of cells within disease foci. Microarray hybridisation glass chips spotted with sets of genes can then be used to obtain a molecular fingerprint of gene expression in the microdissected cells. The variation of expressed genes or alterations in the cellular DNA that correlate with a particular disease state can be compared within or between individual samples. The identification of gene expression patterns may provide vital information for the understanding of the disease process and may contribute to diagnostic decisions and therapies tailored to the individual patient. Molecules found to be associated with defined pathological lesions may provide clues about new therapeutic targets in the future.

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Microdissection and Dop-PCR-Based Reverse Chromosome Painting as a Fast and Reliable Strategy in the Analysis of Various Structural Chromosome Abnormalities

Cited by 21 publications

References 32 publications

Wolf–Hirschhorn syndrome-associated chromosome changes are not mediated by olfactory receptor gene clusters nor by inversion polymorphism on 4p16

Wolf–Hirschhorn syndrome-associated chromosome changes are not mediated by olfactory receptor gene clusters nor by inversion polymorphism on 4p16

Mental and psychomotoric retardation in two brothers with pure partial trisomy 7q32-q34 due to a maternal insertion (14;7)

New developments in the analysis of gene expression

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