BIOLOGICAL-ACTIVITY IN THE REPOPULATING RAT SPERMATOCYTE AFTER THE WITHDRAWAL OF GOSSYPOL TREATMENT .6. ALTERATION IN NUCLEAR FACTORS FOR INTERACTION WITH HISTONE GENE PROMOTER
Cs. Teng et al., BIOLOGICAL-ACTIVITY IN THE REPOPULATING RAT SPERMATOCYTE AFTER THE WITHDRAWAL OF GOSSYPOL TREATMENT .6. ALTERATION IN NUCLEAR FACTORS FOR INTERACTION WITH HISTONE GENE PROMOTER, Contraception, 52(2), 1995, pp. 129-134
This article reports the effects of gossypol at the genomic level in r
at spermatogenic cells. After gossypol treatment for various times (8,
12, and 19 weeks), the spermatogonial cells were allowed to rest for
2 to 4 weeks. The function of histone H-4 gene promoter (H(4)GP) in th
e repopulating pachytene spermatocytes (RPS) was investigated. The seq
uences of the oligonucleotides for the H(4)GP binding sites 2 and 2 we
re synthesized by an ABI-392 DNA synthesizer. RPS and the control pach
ytene spermatocytes (CPS) were obtained by centrifugal elutriation and
subsequently they were used for the preparation of nuclear protein ex
tracts (NPE). The NPE interaction with the DNA fragment of site I or 2
was studied by an electrophoresis mobility shift assay (EMSA). EMSA w
ith NPE-CPS revealed ten major gel shift bands for site I and 2. The p
resence of extra unlabeled DNA fragments competed with 6 of the bands.
After 2 to 4 weeks recovery from 8, 12, and 19 weeks of gossypol trea
tment, NPE-RPS failed to shift four bands (b through e) in site 1. The
se results suggested that gossypol treatment affected the transcriptio
n factors for interaction with site 1. On the contrary, no effect was
demonstrated in NPE that interacted with site 2. Furthermore, gossypol
treatment did not change the nucleotide sequence in the H(4)GP site 1
and 2.