Human interphase chromosomes: a review of available molecular cytogenetic technologies

Abstract

Human karyotype is usually studied by classical cytogenetic (banding) techniques. To perform it, one has to obtain metaphase chromosomes of mitotic cells. This leads to the impossibility of analyzing all the cell types, to moderate cell scoring, and to the extrapolation of cytogenetic data retrieved from a couple of tens of mitotic cells to the whole organism, suggesting that all the remaining cells possess these genomes. However, this is far from being the case inasmuch as chromosome abnormalities can occur in any cell along ontogeny. Since somatic cells of eukaryotes are more likely to be in interphase, the solution of the problem concerning studying postmitotic cells and larger cell populations is interphase cytogenetics, which has become more or less applicable for specific biomedical tasks due to achievements in molecular cytogenetics (i.e. developments of fluorescence in situ hybridization -- FISH, and multicolor banding -- MCB). Numerous interphase molecular cytogenetic approaches are restricted to studying specific genomic loci (regions) being, however, useful for identification of chromosome abnormalities (aneuploidy, polyploidy, deletions, inversions, duplications, translocations). Moreover, these techniques are the unique possibility to establish biological role and patterns of nuclear genome organization at suprachromosomal level in a given cell. Here, it is to note that this issue is incompletely worked out due to technical limitations. Nonetheless, a number of state-of-the-art molecular cytogenetic techniques (i.e multicolor interphase FISH or interpahase chromosome-specific MCB) allow visualization of interphase chromosomes in their integrity at molecular resolutions. Thus, regardless numerous difficulties encountered during studying human interphase chromosomes, molecular cytogenetics does provide for high-resolution single-cell analysis of genome organization, structure and behavior at all stages of cell cycle.

Somatic genomic variations in extra-embryonic tissues

“…This low frequency of genomic variants marks a demand for highly sensitive technologies-of-scale that offer the high throughput necessary to detect such rare events. The application of improved methods for interphase cell analysis the [62][63][64] and novel genomic and proteomic technology platforms that have been developed in recent years promise to meet this need [65]. …”

Genomic Instability in the Brain: Etiology, Pathogenesis and New Biological Markers of Psychiatric Disorders

“…Однако только немногие из них были оцене-ны в свете вариаций генома, проявляющихся как GIN или CIN. Среди них такие моногенные CIN-синдромы, как атаксия-телеангиэктазия, болезнь Альцгеймера и синдром Дауна (трисомия хромосомы 21) [27,33,42,43]. Обнаружено, что возрастание уровня анеуплоидии в онто-генезе, по-видимому, характерно для болезней, ассоции-рованных с преждевременным старением (например, АТ) [18,41], но также наблюдается и в малигнизированных тканях [3,44].…”

To see an interphase chromosome or: How a disease can be associated with specific nuclear genome organization

“…Since the introduction of interphase molecular cytogenetics a significant effort has been made to provide comprehensive information about the meaning of nuclear genome (chromosome) organization [1][2][3][13][14][15]. As a result, interphase chromosome architecture was demonstrated to be involved in critical nuclear processes, which are relevant to cellular homeostasis in health and disease (Box 1).…”

“…As noted below, interphase molecular cytogenetic techniques are usually applied either for analysis of specific genomic loci (using probes for relatively small DNA sequences (rarely >1Mb) comparing to the whole chromosomes) or for painting the whole chromosome, visualized as a chromosome territory (reviewed in [3]). Although these approaches are successfully applied for studying chromosomal numbers and intranuclear arrangement, the impossibility to identify positioning of specific chromosomal regions in relation to the chromosome itself and to other chromosomal regions significantly reduces the resolution of interphase chromosomal analysis [12].…”

“…More precisely, the availability to visualize either specific genomic loci or ambiguous chromosome territories was found to be insufficient for interphase cytogenetics. To solve this problem, a technique providing for simultaneous visualization of the whole chromosome and its regions in a given nucleus has appeared to be required [3]. The development of interphase chromosome-specific multicolor banding (ICS-MCB) has led the way towards the high-resolution interphase cytogenetic analysis for Visualization of interphase chromosomes studying chromosomal numbers, structure and spatial arrangement within the nucleus [4][5][6][7].…”