Single lymphocytes from two healthy individuals with mitochondrial point heteroplasmy are mainly homoplasmic

被引：11

作者：

Lutz-Bonengel, Sabine ^{[1
]}

Saenger, Timo ^{[1
]}

Parson, Walther ^{[2
]}

Mueller, Helena ^{[3
]}

Ellwart, Joachim W. ^{[4
]}

Follo, Marie ^{[5
]}

Bonengel, Bernhard ^{[6
]}

Niederstaetter, Harald ^{[2
]}

Heinrich, Marielle ^{[1
]}

Schmidt, Ulrike ^{[1
]}

机构：

[1] Univ Freiburg, Inst Med Legale, D-79104 Freiburg, Germany

[2] Innsbruck Med Univ, Inst Med Legale, Innsbruck, Austria

[3] Advalytix AG, D-85649 Brunnthal, Germany

[4] GSF Natl Res Ctr Environm & Hlth, Inst Mol Immunol, D-81377 Munich, Germany

[5] Univ Freiburg, Med Ctr, Core Facil, Dept Hematol & Oncol, Freiburg, Germany

[6] HLS Offenburg BTG, D-77652 Offenburg, Germany

来源：

INTERNATIONAL JOURNAL OF LEGAL MEDICINE | 2008年 / 122卷 / 03期

关键词：

mitochondrial DNA; sequence heteroplasmy; single cells; flow cytometry;

D O I：

10.1007/s00414-007-0190-6

中图分类号：

DF [法律]; D9 [法律]; R [医药、卫生];

学科分类号：

0301 ; 10 ;

摘要：

The nature of mitochondrial DNA heteroplasmy is still unclear. It could either be caused by two mitochondrial DNA (mtDNA) haplotypes coexisting within a single cell or by an admixture of homoplasmic cells, each of which contains only one type of mtDNA molecule. To address this question, single lymphocytes were separated by flow cytometry assisted cell sorting and analyzed by cycle sequencing or minisequencing. To attain the required PCR sensitivity, the reactions were carried out on the surface of chemically structured glass slides in a reaction volume of 1-2 mu l. In this study, blood samples from two healthy donors showing mitochondrial point heteroplasmy in direct sequencing (195Y and 234R, respectively) were analyzed. Nearly 96% of single lymphocytes tested were found to be in a homoplasmic state, but heteroplasmic cells were also detected. These results suggest that mitochondrial point heteroplasmy in blood may well be mainly due to the mixture of homoplasmic cells.

引用

页码：189 / 197

页数：9

共 51 条

[1] Results of the 1999-2000 collaborative exercise and proficiency testing program on mitochondrial DNA of the GEP-ISFG:: an inter-laboratory study of the observed variability in the heteroplasmy level of hair from the same donor [J].

Alonso, A ;

Salas, A ;

Albarrán, C ;

Arroyo, E ;

Castro, A ;

Crespillo, M ;

di Lonardo, AM ;

Lareu, MV ;

Cubría, CL ;

Soto, ML ;

Lorente, JA ;

Semper, MM ;

Palacio, A ;

Paredes, M ;

Pereira, L ;

Lezaun, AP ;

Brito, JP ;

Sala, A ;

Vide, MC ;

Whittle, M ;

Yunis, JJ ;

Gómez, J .

FORENSIC SCIENCE INTERNATIONAL, 2002, 125 (01) :1-7

[2] Reanalysis and revision of the Cambridge reference sequence for human mitochondrial DNA [J].

Andrews, RM ;

Kubacka, I ;

Chinnery, PF ;

Lightowlers, RN ;

Turnbull, DM ;

Howell, N .

NATURE GENETICS, 1999, 23 (02) :147-147

[3]

BENDALL KE, 1995, AM J HUM GENET, V57, P248

[4]

Bendall KE, 1996, AM J HUM GENET, V59, P1276

[5] Quantification and sequencing of somatic deleted mtDNA in single cells: evidence for partially duplicated mtDNA in aged human tissues [J].

Bodyak, ND ;

Nekhaeva, E ;

Wei, JY ;

Khrapko, K .

HUMAN MOLECULAR GENETICS, 2001, 10 (01) :17-24

[6] Mitochondrial DNA heteroplasmy or artefacts -: a matter of the amplification strategy? [J].

Brandstätter, A ;

Parson, W .

INTERNATIONAL JOURNAL OF LEGAL MEDICINE, 2003, 117 (03) :180-184

[7] The frequency of heteroplasmy in the HVII region of mtDNA differs across tissue types and increases with age [J].

Calloway, CD ;

Reynolds, RL ;

Herrin, GL ;

Anderson, WW .

AMERICAN JOURNAL OF HUMAN GENETICS, 2000, 66 (04) :1384-1397

[8] The mitochondrial bottleneck occurs without reduction of mtDNA content in female mouse germ cells [J].

Cao, Liqin ;

Shitara, Hiroshi ;

Horii, Takuro ;

Nagao, Yasumitsu ;

Imai, Hiroshi ;

Abe, Kuniya ;

Hara, Takahiko ;

Hayashi, Jun-Ichi ;

Yonekawa, Hiromichi .

NATURE GENETICS, 2007, 39 (03) :386-390

[9] Analysis of mtDNA copy number and composition of single mitochondrial particles using flow cytometry and PCR [J].

Cavelier, L ;

Johannisson, A ;

Gyllensten, U .

EXPERIMENTAL CELL RESEARCH, 2000, 259 (01) :79-85

[10] The inheritance of mitochondrial DNA heteroplasmy: random drift, selection or both? [J].

Chinnery, PF ;

Thorburn, DR ;

Samuels, DC ;

White, SL ;

Dahl, HHM ;

Turnbull, DM ;

Lightowlers, RN ;

Howell, N .

TRENDS IN GENETICS, 2000, 16 (11) :500-505

← 1 2 3 4 5 6 →