Karyotyping and Chromosomal Analysis of Rodents

Eighteen specimens belonging to seven species were karyotyped. Mitotic chromosome suspensions were made in the field from short-term culture of bone marrow and/or spleen after colchicine treatment in vivo as described previously^{44 (link)}. For U. aff. soricipes, a chromosome suspension from primary fibroblast culture was prepared following the standard technique⁴⁵. Air-dried chromosome spreads of all the specimens were conventionally stained with 2% Giemsa for 1–2 min and sequentially subjected to differential staining.
The standard trypsin/Giemsa staining procedure was carried out for the identification of each chromosome arm by G-bands^{46 (link)}. C-banding was performed by the classic technique^{47 (link)}. NORs were detected by silver nitrate staining^{48 (link)}.
A Leica DFC-295 CCD camera mounted on a DM1000 (Leica) microscope or a Metasystems CCD (Zeiss) camera mounted on an AxioScope 2 (Zeiss) microscope was employed to capture images using MetaSystems Ikaros ver.5.3 and Leica Application ver.3.2 software packages, respectively.

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Pavlova S.V., Lebedev V.S., Yakushov V.D., Zhu Y., Fang Y., Sun Y.H, & Sheftel B.I. (2021). High diversity of small insectivorous mammals on Qinghai–Tibet Plateau and first description of karyotype for four endemics of China. Scientific Reports, 11, 24496.

Publication 2021

DM1000 AxioScope 2 Leica Application ver.3.2

Bone marrow Chromosome Colchicine Fibroblast Microscope Nors Silver nitrate Spleen Trypsin

Corresponding Organization : Severtsov Institute of Ecology and Evolution

Other organizations : Lomonosov Moscow State University, Institute of Zoology

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Variable analysis

independent variables

Treatment with colchicine in vivo for preparing mitotic chromosome suspensions
Use of primary fibroblast culture for preparing chromosome suspension for U. aff. soricipes

dependent variables

Chromosome karyotypes of the 18 specimens belonging to 7 species

control variables

Short-term culture of bone marrow and/or spleen for preparing mitotic chromosome suspensions (except for U. aff. soricipes)
Conventional Giemsa staining of chromosome spreads
Trypsin/Giemsa staining for G-banding
C-banding technique
Silver nitrate staining for NOR detection

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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