This study aimed to evaluate the effect of seminal vesicle fluid (SVF) on the acrosome reaction (AR) occurred spontaneously or induced by Ca2+ ionophore A23187, follicular fluid, and progesterone in mouse epididymal sperm. SVF was divided into high (MW>10 kM)) and low (MW<10 kD) fractions by ultrafiltration. The low MW fraction of SVF decreased the rate of spontaneous AR, however the high MW fraction did not. It suggested that the low MW fraction of SVF might have contained decapacitation factor(s) responsible for prolonging of time need for capacitation. When sperm preincubated for 60 min in the presence of SVF, the rate of AR induced by A23187 was decreased, but prolongation of preincubation time for 120 min significantly potentiated the AR by A23187. It suggested that addition of SVF into sperm preincubation medium imposed the epididymal sperm a condition similar to ejaculation. AR induced by human follicular fluid or progesterone was also inhibited by SVF. It suggested that substance in SVF might have affected AR of mouse sperm by inhibiting the interaction between AR inducing ligands and sperm surface receptors involved in acrosomal exocytosis.
Acrosome Reaction* ; Acrosome* ; Animals ; Calcimycin ; Ejaculation ; Exocytosis ; Female ; Follicular Fluid ; Humans ; Ligands ; Male ; Mice* ; Progesterone ; Seminal Vesicles* ; Spermatozoa* ; Ultrafiltration

Stress coping mechanisms are critical to minimize or overcome damage caused by ever changing environmental conditions. They are designed to promote cell survival. The unfolded protein response (UPR) pathway is mobilized in response to the accumulation of unfolded proteins, ultimately in order to regain endoplasmic reticulum (ER) homeostasis. Various elements of coping responses to ER stress including Perk, Ask1, Bip, Chop, Gadd34, Ire1, Atf4, Atf6, and Xbp1 have been identified and were found to be inducible in oocytes and preimplantation embryos, suggesting that, as a normal part of the cellular adaptive mechanism, these coping responses, including the UPR, play a pivotal role in the development of preimplantation embryos. As such, the UPR-associated molecules and pathways may become useful markers for the potential diagnosis of stress conditions for preimplantation embryos. After implantation, ER stress-induced coping responses become physiologically important for a normal decidual response, placentation, and early organogenesis. Attenuation of ER stress coping responses by tauroursodeoxycholate and salubrinal was effective for prevention of cell death of cultured embryos. Further elucidation of new and relevant ER stress coping responses in periimplantation embryos might contribute to a comprehensive understanding of the regulation of normal development of embryonic development and potentiation of embryonic development in vitro.
Blastocyst ; Cell Death ; Cell Survival ; Diagnosis ; Embryonic Development ; Embryonic Structures* ; Endoplasmic Reticulum ; Endoplasmic Reticulum Stress* ; Female ; Homeostasis ; Oocytes ; Organogenesis ; Placentation ; Pregnancy ; Unfolded Protein Response

OBJECTIVE Present study was aimed to verify the effect of granulocyte macrophage-colony stimulating factor (GM-CSF) in the preimplantation development of mouse embryos and the involvement of the mitogen activated protein kiase (MAPK) in the GM-CSF signaling. METHODS: Two-cell embryos were cultured for 96 h in the presence or absence of GM-CSF (0, 0.4, 2, 10 ng/ml) and PD98059, a MEK inhibitor (10 muM). Morphological development, cell number per blastocyst, and apoptotic nuclei, were eamined. MAPK activity of embryonic immunoprecipitate by MAPK (ERK1/2) antibody was measured by in vitro phosphorylation of myelin basic protein. RESULTS: At post hCG 122 h the embryonic development among the experimental groups was significantly different (p=0.018). The rate of blastocyst development and cell number per embryo were the highest in 2 ng/ml GM-CSF treatment group. The percent of apoptotic cells of the GM-CSF-treated embryos was the lowest among the group. in blastocysts, GM-CSF treatment transiently increased MAPK activity. PD098059 attenuated the effect of GM-CSF on the morphological development, increase in cell number per blastocyst, down regulation of apoptosis, and upregulation of MAPK activity, suggesting that activation of MAPK cascade possibly mediated the embryotropic effect of GM-CSF. CONCLUSION: This result suggested that GM-CSF potentiated the development of preimplantation mouse embryos by activation of MAPK.
Animals ; Apoptosis ; Blastocyst ; Cell Count ; Down-Regulation ; Embryonic Development ; Embryonic Structures* ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; Granulocytes* ; Mice* ; Myelin Basic Protein ; Phosphorylation ; Pregnancy ; Up-Regulation

PURPOSE: To verify the regulation of transepithelial resistance (TER) of Sertoli cells by Leydig cells in mouse. MATERIALS AND METHODS: Primary culture of Sertoli cells was established on cell culture plate insert and monolayer culture was subjected to coculture in the Leydig cell culture. Changes in TER was monitored for 48 h using the conductivity meter equipped with two electrodes system. RESULTS: TER gradually increased according to the development of monolayer of Sertoli cells on the cell culture plate insert. Net changes in TER of Sertoli cells culture was significantly higher under the Leydig cells coculture compared to control after 48 h of coculture. CONCLUSIONS: It is the first report about the increase in TER of Sertoli cells by Leydig cells in vitro. Paracrine interaction between Leydig cells and Sertoli cells might be involved in the development of functional blood testis barrier which is made by tight junctions between Sertoli cells in mouse testis.
Animals ; Blood-Testis Barrier ; Cell Culture Techniques ; Coculture Techniques* ; Electrodes ; Leydig Cells* ; Male ; Mice* ; Sertoli Cells* ; Testis ; Tight Junctions

The present study was designed to define the role of prostaglandin in the development and hatching of mouse embryo. The effects of indomethacin, an inhibitor of prostaglandin synthesis, on the development and hatching of morula and blastocyst were examined. In early morula stage, embryos were degenerated significantly at 100 muM and 200 muM indomethacin. However, :he viability of embryos was not influenced by concentration in any other embryonic stages. In all embryonic stages, the hatching was suppressed with concentration dependent manner, but expansion was not suppressed. Particularly, in 84h embryos post hCG injection, the hatching was suppressed significantly compared with post hCG 72h or 96h embryos. When embryos were treated with 100 muM indomethacin for a specific time (12h) in according to the development stage, the hatching was suppressed all groups. These suppressional effect was decreased as embryonic development stage was progressed. However, the expansion was not affected in all treatment group. This study suggests that hatching-related metabolic substances are synthesized from morula stage and intraembryonic signaling mediated prostaglandin was important for development and hatching of mouse embryo.
Animals ; Blastocyst ; Embryonic Development ; Embryonic Structures* ; Female ; Indomethacin* ; Mice* ; Morula ; Pregnancy

No abstract available.
Animals ; Mice* ; Spermatozoa*

No abstract available.
Animals ; Epithelium* ; Mice* ; Oocytes* ; Oviducts* ; Phenobarbital*

PURPOSE: To verify the expression of mouse homologue of glucosamine-6-phosphate deaminase (GNPDA) in mouse epididymis. MATERIALS AND METHODS: GNPDA expression was examined in mouse epididymis using the polyclonal antibody raised against a synthetic oligopeptide of N-terminus of mouse homologue of GNPDA. RESULTS: In Western blot, immunoreactive band of Mr. 33 kDa was detected in the cauda epididymal extracts. In immunostaining, positive signal was found in the cytoplasm and stereocillia of epithelial cells of cauda epididymal tubule, and interstitial cells of adult epididymis. CONCLUSION: Expression of GNPDA in epididymis suggests that it possibly involved in absorption and intracellular digestion of luminal contents, and the remodeling of epithelia in mouse epididymis.
Absorption ; Adult ; Animals ; Blotting, Western ; Cytoplasm ; Digestion ; Epididymis* ; Epithelial Cells ; Humans ; Male ; Mice* ; Phenobarbital

PURPOSE: To verify the expression of calcium-binding proteins in mouse spermatozoa. MATERIALS AND METHODS: Mouse sperm proteins were subjected to 2-dimentional SDS-PAGE combined with calcium shift in the presence or absence of Ca2+, Zn2+ or EDTA. RESULTS: In the epididymal sperm extracts, 10 kinds of protein spots showed mobility shift in the gel containing Ca2+. Most of the CBPs disappeared after the acrosome reaction (AR) induced by Ca2+-ionophore A23187, suggesting that they originated from acrosome and/or the plasma membrane overlaying the acrosome. CONCLUSIONS: The CBPs might be involved in acrosome reaction of mouse spermatozoa. Protein database of sperm CBPs might be useful for diagnosis of male infertility.
Acrosome ; Acrosome Reaction ; Animals ; Calcimycin ; Calcium ; Calcium-Binding Proteins* ; Cell Membrane ; Databases, Protein ; Diagnosis ; Edetic Acid ; Electrophoresis, Polyacrylamide Gel* ; Infertility, Male ; Male ; Mice* ; Spermatozoa*

No abstract available.
Adenosine Triphosphatases* ; Adenosine* ; Alkaline Phosphatase* ; Female ; Ovarian Follicle*