Yolk proteins
during ovary and egg development of mature female freshwater crayfish
(Cherax quadricarinatus)
V. Serrano-Pinto, C. Vazquez-Boucard, H. Villarreal-Colmenares-2003
Comparative Biochemistry and Physiology A
–Molecular & Integrative Physiology 134(1): 35-45
(via Current Contents)
Abstract:
Vitellins
from ovaries and eggs at different stages of development in freshwater
crayfish (Cherax quadricarinatus) were examined by chromatography, PAGE and
SDS-PAGE. With these methods, two forms of vitellin (Vt1 and Vt2) were
observed in ovaries and eggs (stages I and V). In ovaries in secondary
vitellogenesis, native molecular mass was 470 (Vt1) and 440 (Vt2) kDa. The
electrophoretic pattern of the eggs proved to be more complex. The protein
molecular mass depends on the development stage of the egg: stage 1, 650 kDa
(Vt1) and 440 kDa (Vt2); stage V, 390 kDa (Vt1) and 340 kDa (Vt2). The
identified vitellins appear to be lipo-glycocarotenoprotein. A similar
vitellin polypeptide composition was observed in the two forms of vitellin
from ovaries and eggs in stage V. In ovaries the SDS-PAGE analysis showed
four subunits with molecular weights of approximately 180, 120, 95 and 80
kDa (Vt1 and Vt2). The polypeptide composition in the two forms of vitellins
in stage I and stage III eggs were different at 195, 190, 130 and 110 kDa
(Vt1) and 116 and 107 kDa (Vt2). On the other hand, in stage V eggs, 110,
95, 87 and 75 kDa (Vt1 and Vt2) were identified. Two antibodies (Ab1 and
Ab2) were prepared against the purified proteins of stage V eggs and their
specificity was demonstrated by radial immunoprecipitation, and Western
blotting analysis. Two forms of vitellins were also found in stage V eggs
after chromatography on Sepharose CL-2B column and hydroxylapatite and
polyacrylamide gel electrophoresis.
(NW
Ctr. Biol. Res., Dept. Marine Aquaculture Programa Acuacultura, POB 128, La Paz BCS 23000,
Mexico, e-mail: vserrano@cibnor.mx)