1.Ventral Prostate Glands Modification of Sperm Membrane Proteins in Golden Hamster
Jianmin LUO ; Lydia CHENG ; Pakham CHOW
Journal of Sun Yat-sen University(Medical Sciences) 2008;29(3):241-247
[Objective] To investigate the effect of the secretory proteins of the ventral prostate glands on the sperm membrane proteins in golden hamsters. [Methods] The sperm was collected from female hamsters uteri after mated with the males with or without ventral prostate gland. Male golden hamsters were divided into four experimental groups: (i) all accessory sex glands (ASG) removed; (ii) ventral prostate gland removed; (iii) all ASG removed except ventral prostate gland and (iv) sham-operated controls. Each group contained 6 hamsters. The sperm membrane proteins were extracted from uterine sperm and analyzed by SDS-PAGE and two dimension electrophoresis. [Results] The SDS-PAGE results of sperm membrane showed that the ventral prostate glands added up some proteins or increased the quantity of some proteins, which contained molecular mass (MM)15 k, 29 k, 38 k, 55 k, and 91 k proteins, to the sperm membrane. 2D-electrophoresis of sperm membrane showed that some extra protein spots were appeared in with ventral prostate gland groups, their MM and isoelectric point (IP) corresponding to 16 k/8.60, 16.6 k/9.20, 28 k/5.88, 28 k/6.10, 29 k/5.98, 32 k/6.35, 32 k/6.50, 32 k/7.20, 61 k/5.90,and 83 k/6.40, respectively. The quantity of some protein spots increased in sperm membrane of ventral prostate glands group, their MM/IP coresponding to 17 k/5.95, 17.5 k/6.50, 24 k/7.20, 26 k/5.40, 26 k/5.60, 27 k/7.20, 27 k/7.50, 28 k/5.70, 29 k/8.50, 42 k/6.50, and 42 k/7.00, respectively. [Conclusion] The ventral prostate gland secretion plays a role in regulating the sperm membrane proteins and the latter may in turn affect the male fertility or embryo development.
2.Effect of ventral prostate secretory proteins on oviductal fluid glycoproteins in golden hamsters.
Jian-Min LUO ; Lydia CHENG ; Yuan-Cong ZHOU ; Patricia CHOW
National Journal of Andrology 2008;14(1):42-46
OBJECTIVETo investigate the effect of the secretory proteins of the ventral prostate on the glycoproteins in the oviductal fluid of golden hamsters.
METHODSMale golden hamsters were divided into four groups: sham operation (SH), total removal of accessory sex glands (TX), and retainment of the ventral prostate only (VP). Oviductal fluid was collected from female hamsters at 0.5, 2, 4 and 6 h after mating with the males of different operated groups with or without ventral prostate. Glycoproteins were probed with a panel of lectins and their changes in the oviductal fluid were analyzed by Western blot.
RESULTSThe 47 000, 52 000, 81 000 and 128 000 WGA-binding proteins were observed in the oviductal fluid of the 6 h TX group, the 32 000, 35 500, 47 000 and 52 000 WGA-binding glycoproteins noted in the 6 h VP group, the 47 000, 68 000, 95 000 and 128 000 pisum sativum agglutinin (PSA)-binding glycoproteins shown in the 6 h TX and VP groups, two extra 32 000 and 37 500 bands detected in the 6 h VP group, the 47 000 and 52 000 dolichos biflorus agglutinin (DBA)-binding glycoproteins present in the 6 h VP but absent in the 6 h TX group.
CONCLUSIONVentral prostate secretory proteins affect acetylglucosamine, N-acetylgalactosamine/galactose and mannose in the oviductal fluid collected 6 hours after mating. And these glycoproteins may play an important role in the development of embryos.
Animals ; Copulation ; physiology ; Cricetinae ; Fallopian Tubes ; metabolism ; Female ; Glycoproteins ; metabolism ; Male ; Mesocricetus ; Prostatic Secretory Proteins ; physiology
3.Identification of sperm-binding proteins in the ventral prostate of the golden hamster.
Jian-Min LUO ; Lydia CHENG ; Yuan-Cong ZHOU ; O WAI-SUM ; Pak-Ham CHOW
National Journal of Andrology 2007;13(10):883-889
OBJECTIVETo investigate the binding of secretory proteins in the ventral prostate to the surface of sperm.
METHODSWe used different techniques to demonstrate the possibility of ventral prostate secretory proteins binding to sperm in golden hamsters. Polyclonal antibodies against crude secretion of the ventral prostate cultured in rabbits were used to detect the antigens in hamster epididymal, uterine and oviductal spermatozoa by indirect immunofluorescence technique. The uterine and oviductal spermatozoa were collected after mating with the males with or without ventral prostate glands. The ventral prostate secretory proteins were isolated and transblotted to the membrane, which was incubated with the biotinylated epididymal sperm membrane proteins, and then the biotinylated binding proteins were stained.
RESULTSAn immunoreaction restricted to the middle piece was observed in the sperm incubated with the ventral prostate secretion and ejaculated sperm recovered from the uteri and oviducts. The rate of the epididymal sperm bound with the ventral prostate secretory proteins was (80 +/- 5) %, and the rats of the sperm binding to the ventral prostate secretory proteins were (30.0 +/- 4.6) % from the uterus and (16.0 +/- 3.6) % from the oviduct after mating with the males with ventral prostate glands, significantly higher than after mating with those without prostate glands (P < 0.01). Five bands were identified by Western blot analysis in vitro of the ventral prostate secretory proteins incubated with biotinylated epididymal sperm membrane proteins.
CONCLUSIONThe present data indicate that ventral prostate secretory proteins bind to the middle piece of sperm in golden hamsters.
Animals ; Blotting, Western ; Cricetinae ; Epididymis ; metabolism ; Fallopian Tubes ; metabolism ; Female ; Fluorescent Antibody Technique, Indirect ; Male ; Mesocricetus ; Prostate ; metabolism ; Protein Binding ; Seminal Vesicle Secretory Proteins ; metabolism ; Spermatozoa ; metabolism ; Uterus ; metabolism