Mouse anti-fibrous sheath antisera (MAFA) produced by immunizing mice
with purified preparations of human sperm tail fibrous sheath (FS) rea
cted with the principal piece of less than 10% of freshly isolated spe
rmatozoa which were immotile and probably dead. Following their dememb
ranation by detergents or repeated freezing and thawing, all spermatoz
oa were stained. This was also demonstrated on spermatozoa dried onto
slides, but the undiluted xenoantisera showed additional reactivity wi
th the acrosomal zone (AZ). Using immunogold electron microscopy, the
target antigens were ultrastructurally localized to the FS, and a few
spermatozoa showed some reaction at the AZ subacrosomal perinuclear th
eca. Following titration of the antibodies, the anti-AZ-reaction becam
e undetectable at a dilution of 1:20 while their reactivity with the p
rincipal piece continued to a 1:400-dilution. These results indicated
that the xenoantisera probably contained an additional unrelated antib
ody component which reacted with the AZ. Western blotting and staining
of purified FS with MAFA detected seven major protein bands with MW r
anging between 25 kDa and 97.4 kDa. In human testes, the 1:50 diluted
MAFA reacted with sperm tails only, indicating the late expression of
the antigenic determinants during spermatogenesis. MAFA did not react
with oesophagus, stomach, duodenum, ileum, nasal lining tissues, uteru
s, pericardium, pancreas, thyroid gland, or cultured fibroblasts. The
xenoantisera did, however stain the skin epidermis and cultured kerati
nocytes which exhibited filamentous cytoplasmic staining although thei
r target antigens could not be biochemically identified. These results
indicate that the FS proteins express antigenic determinants which ar
e not shared with other cytoskeletal elements within the sperm flagell
um or a variety of somatic tissues.