Heparanase improves mouse embryo implantation.
OBJECTIVE
To improve mouse embryonic implantation by recombinant heparanase supplementation. Heparanase, an endoglycosidase-degrading heparan sulfate proteoglycan, may have a role in embryonic implantation because of its enzymatic, angiogenic, and adhesive properties. Increasing endometrial receptivity could improve one of the most difficult pathologies in human fertility.


METHODS
Comparison between mouse blastocysts obtained after 24-hour incubation of morulae with or without heparanase.


METHODS
Experimental laboratory in a medical center.


METHODS
Mice.


METHODS
Morulae were flushed from CB6F1 female mice and incubated for 24 hours at 37 degrees C in M16 medium supplemented with 0.1 mg/mL heparanase (n = 203), with albumin (n = 60), or with medium alone (n = 258).


METHODS
Blastocysts were evaluated by heparanase immunostaining (n = 10), activity assay (n = 283), and transfer to foster mice uterine horns (n = 228). The number of implantation sites was compared.


RESULTS
Immunostaining demonstrated that heparanase is constitutively expressed in mouse morulae and blastocyts. Heparanase supplementation resulted in increased staining and enzymatic activity in blastocyts. Implantation rates for the heparanase, M16 medium, and albumin groups, were 36.9%, 17.8%, and 20%, respectively (P<.01).


CONCLUSIONS
Heparanase was found to be constitutively expressed by blastocyst-stage embryos. Moreover, the amount of heparanase was markedly increased by incubation of morulae with recombinant heparanase, evaluated by immunostaining and enzymatic activity. Heparanase supplementation resulted in approximately a twofold increase in embryo implantation rate in vivo. Taken together, these data suggest that heparanase is actively involved in embryo implantation.
Revel
Ariel
A
Department of Gynecology, Hadassah-Hebrew Medical Center, Jerusalem, Israel. revel@md.huji.ac.il
Helman
Aharon
A
Koler
Moriah
M
Shushan
Asher
A
Goldshmidt
Orit
O
Zcharia
Eyal
E
Aingorn
Helena
H
Vlodavsky
Israel
I
D000818:Animals; D001757:Blastomeres / Q000187:drug effects / Q000201:enzymology*; D003470:Culture Media / Q000494:pharmacology; D010064:Embryo Implantation / Q000502:physiology*; D005260:Female; D005966:Glucuronidase / Q000378:metabolism* / Q000494:pharmacology; D007150:Immunohistochemistry; D051379:Mice; D009028:Morula / Q000187:drug effects / Q000201:enzymology*; D011247:Pregnancy; D011994:Recombinant Proteins / Q000494:pharmacology; D000099175:Heparanase
D003470:Culture Media; D005966:Glucuronidase; D011994:Recombinant Proteins; D000099175:Heparanase