Detection of cross-sex chimerism in the common marmoset monkey (Callithrix jacchus) in interphase cells using fluorescence in situ hybridisation probes specific for the marmoset X and Y chromosomes

Abstract

Chimerism associated with placental sharing in marmosets has been traditionally analysed using conventional chromosome staining on metaphase spreads or polymerase chain reaction. However, the former technique requires the presence of proliferating cells, whereas the latter may be associated with possible blood cell contamination. Therefore, we aimed to develop a single-cell analysis technique for sexing marmoset cells. We applied fluorescent in situ hybridisation (FISH) to cell nuclei using differentially labelled X and Y chromosome-specific probes. Herein we present the validation of this method in metaphase cells from a marmoset lymphoblastoid cell line, as well as application of the method for evaluation of cross-sex chimerism in interphase blood lymphocytes and haematopoietic bone marrow cells from marmosets of same- and mixed-sex litters. The results show conclusively that haematopoietic cells of bone marrow and leucocytes from blood are cross-sex chimeric when the litter is mixed sex. In addition, single samples of liver and spleen cell suspensions from one individual were tested. Cross-sex chimerism was observed in the spleen but not in liver cells. We conclude that FISH is the method of choice to identify cross-sex chimerism, especially when combined with morphological identification of nuclei of different cell types, which will allow a targeted tissue-specific analysis.

Sibling sex, but not androgens, shapes phenotypes in perinatal common marmosets (Callithrix jacchus)

“…While uncommon in most mammals, marmosets regularly exhibit genetic chimerism. That is, because of inter-mingling of fetal stem cells during prenatal development, marmosets often contain some of their siblings’ genetic material 61–63 . However, levels of chimerism vary across individuals.…”

After short interbirth intervals, captive callitrichine monkeys have higher infant mortality

“…Our results also add to a growing body of knowledge about the complex reproductive strategies of callitrichine monkeys, particularly processes associated with sibling competition. Firstly, during callitrichine pregnancy, the intrauterine environment promotes cell swapping between siblings, resulting in whole-body chimeras ( Patten, 2021 ; Ross et al., 2007 ; Silva et al., 2017 ; Wedi et al., 2016 ). That is, any given monkey likely has chimeric tissues composed of two or more individual genomes (even potentially including in the germline ( Patten, 2021 ; Ross et al., 2007 ), but see ( Wedi et al., 2016 )).…”

“…Firstly, during callitrichine pregnancy, the intrauterine environment promotes cell swapping between siblings, resulting in whole-body chimeras ( Patten, 2021 ; Ross et al., 2007 ; Silva et al., 2017 ; Wedi et al., 2016 ). That is, any given monkey likely has chimeric tissues composed of two or more individual genomes (even potentially including in the germline ( Patten, 2021 ; Ross et al., 2007 ), but see ( Wedi et al., 2016 )). Such chimerism is predicted to alter the balance between cooperation to conflict among siblings (see ( Haig, 1999 ; Patten, 2021 ) for detailed treatments).…”

Developmental and family history‐based analysis of congenital fused labia phenotype in the captive common marmoset (Callithrix jacchus)

“…Common marmoset blood samples were taken from hand‐held, conscious young adult animals (routine procedure in the department) . To investigate the possibility of steroid hormone‐associated differences between CFL and non‐CFL adult females, a standard set of three hormones (total oestrogens, progesterone, and androstenedione) was evaluated in serum samples of a random selection of 11 non‐CFL and 15 CFL animals.…”