An enzyme-linked immunosorbent assay (ELISA) was developed and evaluated to detect equine antisperm antibodies (ASA) in horse serum. Six maiden mares between 12 and 18 mo of age were immunized with stallion sperm cells (SC group, N=2), seminal plasma (SP group, N=2), or phosphate-buffered saline (PBS) as a control (C group, N=2). Horses received a second injection of the same antigen 2 wk after the first. Blood was collected weekly for 10 wk after initial immunization and again at Week 15. Serum ASA levels (IgG and IgA) were measured by ELISA using two assay systems, one containing stallion SC as the plate antigen and another containing SP.
In horses immunized with SC, peak IgG levels were detected by ELISA during Wk 2 and 3 after first injection using either plate antigen. The antibody levels persisted through Week 5 and then slowly declined until Week 15. Horses immunized with SP had IgG levels that did not differ from control horses using either ELISA plate antigen. The only significant elevation in serum IgA ASA occured during Week 5 after initial immunization and only in mares immunized with SC as detected by ELISA using SC as the plate antigen. Attachment of ASA to stallion spermatozoa was confirmed by an indirect immunofluorescence assay. 相似文献
Calluses were induced from immature embryos of an indica type rice and finely dispersed cell suspension cultures were initiated from the callus using modified AA medium (S1 medium). The suspension cultures were maintained alternatively (1–2 passages in each medium) in S1 medium and S2 medium, the latter containing KNO3, NH4NO3, proline and glutamine as nitrogen source. Protoplasts of high quality were isolated form suspension cells cultured in S2 medium supplemented with ABA. Embedding the protoplasts in agarose blocks containing NH4NO3-free modified KM8P(PM1) medium and immersing the blocks in NH4NO3-containing modified KM8P(PM3) medium were most effective for obtaining protoplast division and callus formation. The protoplast-derived calluses were precultured in potato extract-aand/or ABA-containing N6(D1, D2 or D3) media and many embryo-like structures were formed. These structures developed into plantlets after being transferred to N6 differentiation (D4) medium. The regenerated plantlets grew into mature plants and beard seeds normally.Abbreviations AA medium
amino acids based medium
- ABA
abscisic acid
- BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- DF
division frequency
- IAA
indoleacetic acid
- KIN
kinetin
- NAA
naphthaleneacetic acid
- PE
planting efficiency 相似文献