Difference in constitutive heterochromatin behaviour between human amniocytes and lymphocytes detected by a sequential in situ exonuclease III digestion-random primer extension procedure.
Fixed chromosomes from human amniotic fluid cells and peripheral blood lymphocytes were digested in situ with exonuclease III and the single stranded DNA obtained was used as template for an in situ random primer extension. Under these conditions an R banding pattern, more evident in lymphocytes than in amniocytes, was obtained. Nevertheless, constitutive heterochromatin of chromosomes 1, 16, Yq, and mainly the pericentromeric region of chromosome 9 was far more intensely labelled in amniocytes than in lymphocytes. Fluorescence in situ hybridisation with a specific classical satellite DNA probe, showed that this differential labelling was dependent on a greater sensitivity of chromosome 9 constitutive heterochromatin to exonuclease III digestion in amniocytes than in lymphocytes, thus indicating qualitative differences in this region between both human cellular materials.
Fernández
J L
JL
Centro Oncológico de Galicia, La Coruña, Spain.
Campos
A
A
López-Fernández
C
C
Gosálvez
J
J
Goyanes
V
V
D000293:Adolescent; D000328:Adult; D000653:Amniotic Fluid / Q000737:chemistry / Q000166:cytology*; D002454:Cell Differentiation; D002648:Child; D002675:Child, Preschool; D002871:Chromosome Banding; D002878:Chromosomes, Human, Pair 1; D002885:Chromosomes, Human, Pair 16; D002899:Chromosomes, Human, Pair 9; D017931:DNA Primers; D004277:DNA, Single-Stranded / Q000235:genetics; D005090:Exodeoxyribonucleases; D005260:Female; D006570:Heterochromatin / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007223:Infant; D007231:Infant, Newborn; D008214:Lymphocytes / Q000737:chemistry*; D008297:Male; D009928:Organ Specificity; D013379:Substrate Specificity; D014998:Y Chromosome
D017931:DNA Primers; D004277:DNA, Single-Stranded; D006570:Heterochromatin; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III