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, 10 (4), 731-752

Comparison of Different Cytogenetic Methods and Tissue Suitability for the Study of Chromosomes in Cimex lectularius (Heteroptera, Cimicidae)


Comparison of Different Cytogenetic Methods and Tissue Suitability for the Study of Chromosomes in Cimex lectularius (Heteroptera, Cimicidae)

David Sadílek et al. Comp Cytogenet.


In the article we summarize the most common recent cytogenetic methods used in analysis of karyotypes in Heteroptera. We seek to show the pros and cons of the spreading method compared with the traditional squashing method. We discuss the suitability of gonad, midgut and embryo tissue in Cimex lectularius Linnaeus, 1758 chromosome research and production of figures of whole mitosis and meiosis, using the spreading method. The hotplate spreading technique has many advantages in comparison with the squashing technique. Chromosomal slides prepared from the testes tissue gave the best results, tissues of eggs and midgut epithelium are not suitable. Metaphase II is the only division phase in which sex chromosomes can be clearly distinguished. Chromosome number determination is easy during metaphase I and metaphase II. Spreading of gonad tissue is a suitable method for the cytogenetic analysis of holokinetic chromosomes of Cimex lectularius.

Keywords: holokinetic chromosomes; karyogram; midgut; spreading method; squashing method; testes.


Figure 1.
Figure 1.
Adult and 5th instar larva Cimex lectularius gonads. A Adult testes B Adult ovaries, without eggs C Adult ovaries, with well-developed eggs D 5th instar larva testes, well-developed, probably sub adult specimen E 5th instar larva ovaries. Scale bar = 1 mm.
Figure 2.
Figure 2.
Various stages of mitotic and meiotic Cimex lectularius chromosomes with basic karyotype 2n = 26+X1X2Y (A, B, D–L) and karyotype 2n = 26+X1-10Y (C), stained with Giemsa. A Mitotic prometaphase ♂ B Mitotic metaphase ♂ C Metaphase I ♂ D Leptotene ♀ E Pachytene ♀ F Diffuse stage ♂ G Diffuse stage - postpachytene transition ♂ H Postpachytene ♂ I Late postpachytene ♂ J Prometaphase I ♂ K Metaphase I ♂ L Metaphase II ♂. Arrow = sex chromosome (F, G, L) or fragments of supposedly sex chromosomes (C). Scale bar = 5 μm.
Figure 3.
Figure 3.
Male mitotic and meiotic karyograms of Cimex lectularius chromosome variants. A-C Mitotic prometaphase. A 2n = 26+X1X2Y B 2n = 26+X1-6Y C 2n = 26+X1-10Y D Prometaphase II, 2n = 26+X1X2Y E Metaphase II, 2n = 26+X1-7Y. Scale bar = 5 µm.

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