Immunolocalization of actin in intact and DNA- and histone-depleted nuclei and chromosomes of allium cepa.
The presence of actin in eukaryotic nuclei and chromosomes, and especially in higher plant nuclei and chromosomes, has not been well established. We detected actin in meristematic cells of Allium cepa with indirect immunofluorescence technique and observed bright fluorescence in the intact nuclei and chromosomes, indicating that actin is present in the nuclei and chromosomes of the higher plant. We labeled sections of the meristematic cells of A. cepa with immunogold technique, gold particles were found over the whole nuclei and a number of gold particles were concentrated in condensed chromatin and nucleoli, confirming the results of the immunofluoresence observations. We treated the nuclei and chromosomes of A. cepa with DNase I and 2M NaCl and obtained DNA- and histone-depleted nuclei and chromosomes. Indirect immunofluorescence tests showed that the DNA- and histone-depleted nuclei and chromosomes reacted positively with the anti-actin antibodies. These results demonstrate that actin exists not only in intact nuclei and chromosomes, but also in DNA- and histone-depleted nuclei and chromosomes of the plant. In addition, our immuno-fluorescence tests indicate that tropomyosin is present in the nuclei and chromosomes of A. cepa.
Wan
L H
LH
Institute of Genetics and Cytology, Northeast Normal University, Changchun, China.
Xing
M
M
D000199:Actins / Q000032:analysis*; D000490:Allium / Q000737:chemistry*; D002467:Cell Nucleus / Q000737:chemistry*; D002875:Chromosomes / Q000737:chemistry*; D004247:DNA; D003850:Deoxyribonuclease I; D019084:Fluorescent Antibody Technique, Indirect; D006657:Histones; D007150:Immunohistochemistry; D018519:Meristem / Q000737:chemistry; D012965:Sodium Chloride / Q000494:pharmacology; D014335:Tropomyosin / Q000032:analysis
D000199:Actins; D006657:Histones; D014335:Tropomyosin; D012965:Sodium Chloride; D004247:DNA; D003850:Deoxyribonuclease I