1Reptiles show a remarkable diversity of sex determination mechanisms and sex chromosome 2 systems, derived from different autosomal pairs. The origin of the ZW sex chromosomes of 3 Lacerta agilis, a widespread Eurasian lizard species, is a matter of discussion: is it a small 4 macrochromosome from the 11-18 group, common to all lacertids, or this species has unique ZW 5 pair derived from the large chromosome 5. Using independent molecular cytogenetic methods, 6 we investigated the karyotype of L. agilis exigua from Siberia, Russia, to identify the sex 7 chromosomes. FISH with the flow-sorted chromosome painting probe, derived from L. strigata 8 and specific to chromosomes 13, 14, and Z, confirmed that the Z chromosome of L. agilis is a 9 small macrochromosome, the same as in L. strigata. FISH with the telomeric probe showed an 10 extensive accumulation of the telomeric repeat on the W chromosome in agreement with 11 previous studies, excluding the possibility that the lineages of L. agilis studied in different works 12 could have different sex chromosome systems due to a putative intra-species polymorphism. Our 13 results reinforce the idea of the stability of the sex chromosomes and lack of evidence for sex-14 chromosome turnovers in known species of Lacertidae.
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