NOR expression increases in interphase lymphocytes of Down syndrome babies/children as AgNORs surface, according to the mitogen concentration in the culture medium.
The extra chromosome 21 of Down syndrome (DS) or trisomy 21 patients contains an average of 40 extra copies of rRNA genes and the in vivo/in vitro regulation of the activity of these genes is not fully understood. The objective of this work was to compare the NORs expression pattern in interphase lymphocytes of DS patients with regular trisomy 21 and control individuals according to phytohemagglutinin (PHA) concentration (0.37, 0.75, 1.48 and 2.21 ml) per 100 ml of medium. Because the AgNOR staining is an indicator of the active rRNA genes, comparison of the image analysis values of the AgNOR area in 72 h cultivated lymphocytes for each concentration of PHA between DS patients (N=30) and controls (N=24) provided a plausible conclusion on the regulation of the extra rRNA genes in DS lymphocytes. The nucleolus organizer regions area/total nuclear area (NORa/TNa) was calculated using an in-house computer program. Fifty consecutive interphases per PHA concentration were analysed for each individual, for determination of the NORa/TNa. In contrast to healthy controls, NORa/TNa of lymphocytes from DS patient babies/children (0-8 years old) increased gradually in parallel with the PHA concentration in the culture medium: 10.44+/-1.72% for 0.37 ml of PHA, 11.74+/-1.93% for 0.75 ml of PHA, 13.25+/-2.03% for 1.48 ml of PHA and 13.43+/-2.08% for 2.21 ml of PHA per 100 ml of medium. Contrary to control cells (in which the NORa/TNa ratio according to PHA concentration in the culture medium remains constant), DS interphase lymphocytes in culture do not down-regulate their NOR expression. These results obtained from interphase NORs are consistent with the previous results obtained by evaluating the mean of AgNOR+ chromosome number in metaphase cells, also in relation to the mitogen concentration in the culture medium.
Imamoglu
Nalan
N
Department of Medical Biology, Medical Faculty, Erciyes University, 38039 Kayseri, Turkey. nimamoglu@erciyes.edu.tr
Demirtas
Halil
H
Ilten
Ali
A
D034961:Antigens, Nuclear / Q000235:genetics*; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D004314:Down Syndrome / Q000097:blood* / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D020459:Genes, rRNA; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007223:Infant; D007231:Infant, Newborn; D007399:Interphase; D008214:Lymphocytes / Q000502:physiology*; D008934:Mitogens / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics*; D009697:Nucleolus Organizer Region / Q000235:genetics*; D010835:Phytohemagglutinins / Q000494:pharmacology; D013194:Staining and Labeling
D034961:Antigens, Nuclear; D008934:Mitogens; D009687:Nuclear Proteins; D010835:Phytohemagglutinins; C051963:nucleolar organizer region associated proteins