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Akio Miyamoto

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Armando Marques-Guedes

UNL - New University of Lisbon

Case Western Reserve University

Santiago Arango Muñoz

Universidad de Antioquia

DEBAPRASAD BANDYOPADHYAY

Saint-Petersburg State University

MARIA KRITSEPI-KONSTANTINOU

Aristotle University of Thessaloniki

Vietnam National University, Hanoi

Abdulrahman Bello (DVM, MSc, PhD)

Usmanu Danfodio University, Sokoto

University of Sarajevo

Dr. Asmatullah Kaka

Sindh Agriculture University Tandojam-Pakistan

Muhammad Usman Mehmood

University Of Veterinary And Animal Sciences Lahore

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Papers by Akio Miyamoto

Involvement of the luteinizing hormone surge in the regulation of ovary and oviduct clock gene expression in mice

Genes to Cells, Jun 19, 2018

Circadian dysfunction perturbs the female reproductive cycle. In particular, mice lacking the clo... more Circadian dysfunction perturbs the female reproductive cycle. In particular, mice lacking the clock gene Bmal1 show severe infertility, implying that BMAL1 plays roles in ovulation and luteinization. Here, we examined temporal changes in clock gene expression in the ovary and oviduct before and during gonadotropin-induced follicular growth, ovulation, and luteinization in sexually immature mice. While the oviduct did not show a drastic change in clock gene expression, Bmal1 expression in the ovary was higher than that in control mice during the period from 4 to 16 hr after human chorionic gonadotropin (hCG) administration. Bmal1 expression reached a maximum at 16 hr after hCG administration, when follicle luteinization occurred. In an interesting manner, administration of hCG to ex vivo-cultured oviduct triggered a shorter circadian period and inevitably resulted in phase advance. Together, our present data suggest that LH surge induces continuous expression of BMAL1 in the mouse ovary and modulates circadian phase in the mouse oviduct.

Short Communication: A Positive Relationship Between the First Ovulation Postpartum and the Increasing Ratio of Milk Yield in the First Part of Lactation in Dairy Cows

Journal of Dairy Science, May 1, 2007

Changes in Intracellular Calcium Concentrations of in vitro Inseminated and Artificially Activated Bovine Oocytes

Journal of Reproduction and Development, 1996

Expression of 11beta-hydroxysteroid dehydrogenases in bovine follicle and corpus luteum

Journal of Endocrinology, Jun 1, 2003

In glucocorticoid target organs, local concentrations of active glucocorticoid are determined by ... more In glucocorticoid target organs, local concentrations of active glucocorticoid are determined by the relative expression of two 11-hydroxysteroid dehydrogenases (HSDs): bi-directional 11-HSD type1 (11HSD1) that mainly activates cortisone to cortisol, and dehydrogenase 11-HSD type2 (11HSD2) that inactivates cortisol to cortisone. In this study, we examined the expression of mRNA encoding these two 11-HSDs in bovine granulosa cells harvested from preovulatory follicles and corpora lutea (CL). Ovaries were obtained from Holstein cows at a local slaughterhouse. Follicles larger than 10 mm in diameter and CL were dissected and follicular fluid and granulosa cells were taken. Corpora lutea were weighed and their stages were morphologically assessed (stage I, days 1-4; stage II, days 5-10; stage III, days 11-17; stage IV, days 8-20). Follicles were classified into four groups according to their hormonal status (oestradiol (E 2): progesterone (P 4)>1: oestrogen active; E 2 :P 4 <1: oestrogen inactive) and stage of the oestrous cycle (luteal or follicular phase). Total RNA was extracted with phenol-chloroform and subjected to a semi-quantitative RT-PCR for 11HSD1, 11HSD2 and-actin. Concentrations of steroids in follicular fluid were determined by an enzyme immunoassay. In granulosa cells, only 11HSD1 mRNA was detected. There was a negative correlation between the expression of 11HSD1 and the concentration of cortisol in follicular fluid (P<0•05), indicating 11HSD1 may act as a dehydrogenase in the bovine follicle. Both types of 11-HSDs were expressed in CL. The levels of mRNA for both isozymes were high in stage I and II, and were decreased in stage III CL. In stage IV CL, the expression of 11HSD2 but not 11HSD1 mRNA increased. These results indicate that the bovine granulosa cells and CL express 11HSD1 and 11HSD2, and they may play an important physiological role in the bovine ovary through modulating the local glucocorticoid environment.

Evaluation of Teat Skin Cleanliness During Milking at a Dairy Farm Using an ATP-Bioluminescence Assay

Journal of the Japan Veterinary Medical Association, 2013

Intraluteal Release of Prostaglandin F2.ALPHA. and E2 During Corpora Lutea Development in the Cow

Journal of Reproduction and Development, 2002

It is well known that prostaglandin (PG) F2α and PGE2 are actively produced by bovine early corpu... more It is well known that prostaglandin (PG) F2α and PGE2 are actively produced by bovine early corpus luteum (CL) at higher levels than in the CL of later stages. However, there is no in vivo information about the local secretion of PGs within bovine developing CL so far. Thus, the objective of the present study was to determine in detail the real-time changes in PGF 2α and PGE2 secretion within developing CL. A microdialysis system (MDS) was surgically implanted into the newly formed CL of 6 cows on Day 3 after GnRH administration (about 1.5 days after ovulation) to induce superovulation following FSH treatment. Both PGF 2α and PGE2 release were high during the first 24 h within developing CL but decreased thereafter. These profiles were well reflected in the PG concentrations in ovarian venous plasma. Furthermore, reverse transcription polymerase chain reaction analysis using CL from other animals independent from the MDS study revealed that the level of cyclo-oxyganase-2 mRNA expression in early CL was higher than that of later luteal phases. The present results provide the first direct in vivo evidence that active PGs production in CL occurs up to Day 4 after estrus in the cow.

Areolae of the Placenta in the Antarctic Minke Whale (<i>Balaenoptera bonaerensis</i>)

Journal of Reproduction and Development, 2014

In this study, we examined the existence and structure of areolae and the steroidogenesis of areo... more In this study, we examined the existence and structure of areolae and the steroidogenesis of areolar trophoblast cells in the Antarctic minke whale placenta morphologically and immunohistochemically. Placentas were collected from the 15th, 16th and 18th Japanese Whale Research Program under Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. The opening and cavity of fetal areolae formed by taller columnar trophoblast cells (areolar trophoblast cells) with long microvilli and a bright cytoplasm, as compared with the trophoblast cells of the chorionic villi interdigitating with the endometrial crypts, were recognized in observations of serial sections. The opening of the areolar cavity was hidden by chorionic villi with areolar trophoblast cells. Furthermore, a closed pouch-like structure lined by tall columnar cells similar to areolar trophoblast cells within the stroma of chorionic villi was noticed and continued to the areolar cavity, with the opening seen on serial sections. In a surface investigation of the chorion and endometrium by SEM, maternal (endometrial) areolae irregularly surrounded by endometrial folds were obvious. Moreover, we distinguished areolar trophoblast cells with long microvilli attached with many blebs from trophoblast cells. In our immunohistochemical observations, a steroidogenic enzyme, cytochrome P450 side chain cleavage enzyme (P450scc), was detected with strong immunoreactivity in trophoblast cells. However, areolar trophoblast cells showed weak or no immunoreactivity for P450scc.

Hyaluronan regulates sperm-induced inflammatory response by enhancing sperm attachment to bovine endometrial epithelial cells via CD44: in-silico and in-vitro approaches

Frontiers in Endocrinology

Recently, we reported that sperm induce cluster of differentiation 44 (CD44) expression and Toll-... more Recently, we reported that sperm induce cluster of differentiation 44 (CD44) expression and Toll-like receptor 2 (TLR2)-mediated inflammatory response in bovine uterus. In the present study, we hypothesized that the interaction between CD44 of bovine endometrial epithelial cells (BEECs) and hyaluronan (HA) affects sperm attachment and thereby enhancing TLR2-mediated inflammation. To test our hypothesis, at first, in-silico approaches were employed to define the binding affinity of HA for CD44 and TLR2. Further, an in-vitro experiment using the sperm-BEECs co-culture model was applied to investigate the effect of HA on sperm attachment and inflammatory response. Here, low molecular weight (LMW) HA at different concentrations (0, 0.1, 1, or 10 µg/mL) was incubated with BEECs for 2 h followed by the co-culture without- or with non-capacitated washed sperm (106/ml) for additional 3 h was performed. The present in-silico model clarified that CD44 is a high-affinity receptor for HA. Moreo...

Activation of sperm Toll-like receptor 2 induces hyperactivation to enhance the penetration to mucus and uterine glands: a trigger for the uterine inflammatory cascade in cattle

Frontiers in Immunology, Dec 18, 2023

Activation of sperm Toll-like receptor 2 induces hyperactivation to enhance the penetration to mu... more

ヤギ精巣網液および精巣上体液中のインヒビン活性について

Nihon Chikusan Gakkaiho, 1987

Seasonal changes in inhibin activity in seminal plasma and serum concentrations of FSH, LH and testosterone in the male goat ()

Theriogenology, 1987

Inhibin activity in goat seminal plasma was measured by in vitro assay throughout successive 9 mo... more Inhibin activity in goat seminal plasma was measured by in vitro assay throughout successive 9 months and its relationship with the serum FSH, LH and testosterone concentrations was investigated. Total inhibin activity (TIA) in seminal plasma gradually increased from spring to summer, reduced in autumn (P<O.O5) and recovered toward winter (P<O.O5). Serum FSH and LH reached a peak in midsummer (P<O.Ol) and returned to the low levels in autumn. Serum testosterone also increased in midsummer and kept the high levels until the early winter (P<O. 05). Some positive correlation was found in monthly levels between seminal TIA and serum FSH (r=0.305; P<O.O5). Results suggest that the summer increase of inhibin activity in seminal plasma relates with the midsummer rise of serum FSH levels in the male goat.

Evidence that oviduct-sperm-immune cells interaction ensures Th2 environment in bovine oviduct immunity for protecting sperm survival

Zearalenone interferes with the sperm-triggered inflammation in the bovine uterus in vitro: Negative impact on sperm motility and survival

Reproductive Toxicology, 2022

Zearalenone (ZEN)-contaminated diets induce detrimental effects on the bovine reproduction. Recen... more Zearalenone (ZEN)-contaminated diets induce detrimental effects on the bovine reproduction. Recently, we reported that active sperm induce pro-inflammatory responses in bovine endometrial epithelial cells (BEECs) in vitro. This study aimed to investigate the impact of presence of ZEN on the sperm-uterine crosstalk in vitro. BEECs monolayers were stimulated by ZEN (10, 100, and 1000 ng/mL) for 0, 3, 6, 12, or 24 h and gene expressions were analyzed by real-time PCR. Moreover, BEECs were pre-exposed to ZEN (10, 100, and 1000 ng/mL) for 24 h then, co-incubated with sperm for 6 h. Conditioned media (CM) from a sperm-BEECs co-culture, after pre-exposure to ZEN, were harvested and exploited to challenge either polymorphonuclear cells (PMNs) or sperm. Both PMNs phagocytic activity toward sperm and sperm motility parameters were then assessed. Results showed that ZEN alone induced pro-inflammatory responses in BEECs through the induction of mRNA expressions of pro-inflammatory cytokines (TNFA and IL1B) and PGES1 at different time points. Pre-exposure of BEECs to ZEN, amplified the sperm-triggered upregulation of pro-inflammatory cytokines (TNFA and IL1B) and chemokine IL8 mRNA abundance in BEECs. Sperm-BEECs conditioned media, primed by ZEN, stimulated the PMNs phagocytosis for sperm whereas suppressed sperm motility parameters. Taken together, these findings indicate that the presence of ZEN augments the pro-inflammatory cascade triggered by sperm in BEECs, provokes PMNs phagocytosis for sperm, and reduces sperm motility parameters. Such immunological reactions may create a hostile environment for sperm competence and survival in the bovine uterus, thus impair fertility.

Intrauterine infusion of low levels of interferon‐tau on day‐8 post‐estrus stimulates the bovine endometrium to secrete apolipoprotein‐A1: A possible implication for early embryo tolerance

American Journal of Reproductive Immunology, Jul 10, 2022

Toll-like receptor 2 of sperm is involved in sperm attachment to bovine oviductal epithelial cells

Sperm TLR2 regulates Ca2+ influx and acrosome reaction to facilitate sperm penetration to oocytes during in vitro fertilization in cows

An acute-phase protein, alpha 1-acid glycoprotein, is a local regulator to protect sperm from neutrophil phagocytosis in the bovine oviduct

The oviduct is classically described as a sterile milieu, though pathogens and endotoxins could i... more The oviduct is classically described as a sterile milieu, though pathogens and endotoxins could invade its mucosal surfaces via the uterus, peritoneal cavity and follicular fluid. We have recently shown that polymorphonuclear neutrophils PMNs , the first line of defense against microorganisms, are present in the bovine oviduct fluid during preovulatory stages. We further suggested that the bovine oviduct provides a PGE-rich microenvironment to protect sperm from phagocytosis by PMNs that they possibly face in vivo, thereby supporting sperm survival in the oviduct. Alpha 1-acid glycoprotein AGP is a major acute-phase protein produced mainly in the liver, having an immunomodulatory functions. In this study, investigated, 1 the local production of AGP in the bovine oviduct, 2 the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production, and 3 the impact of AGP desialylation on the PMN phagocytosis of sperm. AGP gene was expressed in cultured bovine oviduct epithelial cells BOECs and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps NETs for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to secrete PGE2, and AGP and PGE2 at local levels additively inhibited sperm phagocytosis. The results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.

Evidence that bovine oviduct secrets AGP (a1-acid glycoprotein) to favor a Th2 response and to suppress sperm phagocytosis by neutrophils

Peptidoglycan disrupts early embryo-maternal crosstalk via suppression of ISGs expression induced by interferon-tau in the bovine endometrium

Biochemical and Biophysical Research Communications, Oct 1, 2020

Uterine infection with bacteria and the release of peptidoglycan (PGN), antigenic cell wall compo... more Uterine infection with bacteria and the release of peptidoglycan (PGN), antigenic cell wall components of both Gram-negative and Gram-positive bacteria, can cause early pregnancy losses in ruminants, but the associated mechanisms remain unsolved. Day 7 blastocyst starts to secrete a minute amount of interferon-tau (IFNT) in the uterine horn which is required for early stage of maternal recognition of pregnancy (MRP) in ruminants, and it induces interferon stimulated genes (ISGs) for driving uterine receptivity in cows. This study investigated if PGN disrupts IFNT response through modulation of endometrial ISGs expressions. Cultured bovine endometrial epithelial cells (BEECs) were treated with embryo culture medium (ECM) or IFNT (1 ng/ml) in the presence or absence of a low level of PGN (10 pg/ ml) for 24 h. A real-time PCR analyses revealed that the presence of PGN suppressed IFNT-induced ISGs (OAS1 and ISG15) and STAT1 expressions in BEECs. To visualize the impact of PGN in an ex-vivo model that resembles the in vivo status, endometrial explants were treated by IFNT (1 ng/ml) with or without PGN (10 pg/ml) for 12 h. PGN suppressed IFNT-induced gene expressions of the above factors, but not for IFNA receptor type1 (IFNAR1) or type2 (IFNAR2) in explants. Immunofluorescence analysis illustrated that PGN completely suppressed the IFNT-triggered OAS1 protein expression in the luminal epithelium of explants. Of note, PGN did not stimulate pro-inflammatory cytokines (TNFA and IL1B) or TLR2 mRNA expression in both models. These findings indicate that the presence of low levels of PGN suppresses ISGs expression induced by IFNT secreted from early embryo, at the luminal epithelium of the bovine endometrium. This could severely interfere with early stage of MRP processes in cows, leading to pregnancy failure.

Bovine oviduct epithelial cells suppress the phagocytic activity of neutrophils towards sperm but not for bacteria in vitro: Immunofluorescence and electron microscopic observations

Histology and Histopathology, Oct 17, 2019

Previously, we reported that polymorphonuclear neutrophils (PMNs) are constantly existent in the ... more Previously, we reported that polymorphonuclear neutrophils (PMNs) are constantly existent in the bovine oviduct fluid during the pre-ovulatory stage under physiological conditions. Moreover, incubation of PMNs with bovine oviduct epithelial cells-conditioned medium (BOEC-CM) resulted in suppression of their phagocytic activity for sperm. During pathophysiological conditions, cows may be inseminated by infected semen which exposes oviductal PMNs to allogenic sperm simultaneously with pathogens. This study aimed to visually investigate the role of oviduct epithelium in regulating the phagocytic behavior of PMNs toward sperm as a physiological stimulus, with Escherichia coli (E. coli) as a pathological stimulus. In our experiment, PMNs were incubated for 2 h in BOEC-CM. Phagocytosis was then assayed by co-incubation of these PMNs either with sperm, E. coli, or latex beads. BOEC-CM significantly suppressed the direct phagocytosis of PMNs for sperm, but did not affect their phagocytic activity for E. coli or latex beads. Additionally, an investigation with scanning electron microscopy revealed that BOEC-CM suppressed the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. BOEC-CM did not alter NETs formation towards E. coli. A quantification of NETs formation using an immunofluorescence microscopy showed that the areas of NETs formation for E. coli were significantly larger than those formed for sperm. Our data clearly show that the bovine oviduct, through secretions, protects sperm from phagocytosis by PMNs and eliminates bacterial dissemination through maintaining the phagocytic activity of PMNs towards bacteria.

Involvement of the luteinizing hormone surge in the regulation of ovary and oviduct clock gene expression in mice

Genes to Cells, Jun 19, 2018

Circadian dysfunction perturbs the female reproductive cycle. In particular, mice lacking the clo... more Circadian dysfunction perturbs the female reproductive cycle. In particular, mice lacking the clock gene Bmal1 show severe infertility, implying that BMAL1 plays roles in ovulation and luteinization. Here, we examined temporal changes in clock gene expression in the ovary and oviduct before and during gonadotropin-induced follicular growth, ovulation, and luteinization in sexually immature mice. While the oviduct did not show a drastic change in clock gene expression, Bmal1 expression in the ovary was higher than that in control mice during the period from 4 to 16 hr after human chorionic gonadotropin (hCG) administration. Bmal1 expression reached a maximum at 16 hr after hCG administration, when follicle luteinization occurred. In an interesting manner, administration of hCG to ex vivo-cultured oviduct triggered a shorter circadian period and inevitably resulted in phase advance. Together, our present data suggest that LH surge induces continuous expression of BMAL1 in the mouse ovary and modulates circadian phase in the mouse oviduct.

Short Communication: A Positive Relationship Between the First Ovulation Postpartum and the Increasing Ratio of Milk Yield in the First Part of Lactation in Dairy Cows

Journal of Dairy Science, May 1, 2007

Changes in Intracellular Calcium Concentrations of in vitro Inseminated and Artificially Activated Bovine Oocytes

Journal of Reproduction and Development, 1996

Expression of 11beta-hydroxysteroid dehydrogenases in bovine follicle and corpus luteum

Journal of Endocrinology, Jun 1, 2003

In glucocorticoid target organs, local concentrations of active glucocorticoid are determined by ... more In glucocorticoid target organs, local concentrations of active glucocorticoid are determined by the relative expression of two 11-hydroxysteroid dehydrogenases (HSDs): bi-directional 11-HSD type1 (11HSD1) that mainly activates cortisone to cortisol, and dehydrogenase 11-HSD type2 (11HSD2) that inactivates cortisol to cortisone. In this study, we examined the expression of mRNA encoding these two 11-HSDs in bovine granulosa cells harvested from preovulatory follicles and corpora lutea (CL). Ovaries were obtained from Holstein cows at a local slaughterhouse. Follicles larger than 10 mm in diameter and CL were dissected and follicular fluid and granulosa cells were taken. Corpora lutea were weighed and their stages were morphologically assessed (stage I, days 1-4; stage II, days 5-10; stage III, days 11-17; stage IV, days 8-20). Follicles were classified into four groups according to their hormonal status (oestradiol (E 2): progesterone (P 4)>1: oestrogen active; E 2 :P 4 <1: oestrogen inactive) and stage of the oestrous cycle (luteal or follicular phase). Total RNA was extracted with phenol-chloroform and subjected to a semi-quantitative RT-PCR for 11HSD1, 11HSD2 and-actin. Concentrations of steroids in follicular fluid were determined by an enzyme immunoassay. In granulosa cells, only 11HSD1 mRNA was detected. There was a negative correlation between the expression of 11HSD1 and the concentration of cortisol in follicular fluid (P<0•05), indicating 11HSD1 may act as a dehydrogenase in the bovine follicle. Both types of 11-HSDs were expressed in CL. The levels of mRNA for both isozymes were high in stage I and II, and were decreased in stage III CL. In stage IV CL, the expression of 11HSD2 but not 11HSD1 mRNA increased. These results indicate that the bovine granulosa cells and CL express 11HSD1 and 11HSD2, and they may play an important physiological role in the bovine ovary through modulating the local glucocorticoid environment.

Evaluation of Teat Skin Cleanliness During Milking at a Dairy Farm Using an ATP-Bioluminescence Assay

Journal of the Japan Veterinary Medical Association, 2013

Intraluteal Release of Prostaglandin F2.ALPHA. and E2 During Corpora Lutea Development in the Cow

Journal of Reproduction and Development, 2002

It is well known that prostaglandin (PG) F2α and PGE2 are actively produced by bovine early corpu... more It is well known that prostaglandin (PG) F2α and PGE2 are actively produced by bovine early corpus luteum (CL) at higher levels than in the CL of later stages. However, there is no in vivo information about the local secretion of PGs within bovine developing CL so far. Thus, the objective of the present study was to determine in detail the real-time changes in PGF 2α and PGE2 secretion within developing CL. A microdialysis system (MDS) was surgically implanted into the newly formed CL of 6 cows on Day 3 after GnRH administration (about 1.5 days after ovulation) to induce superovulation following FSH treatment. Both PGF 2α and PGE2 release were high during the first 24 h within developing CL but decreased thereafter. These profiles were well reflected in the PG concentrations in ovarian venous plasma. Furthermore, reverse transcription polymerase chain reaction analysis using CL from other animals independent from the MDS study revealed that the level of cyclo-oxyganase-2 mRNA expression in early CL was higher than that of later luteal phases. The present results provide the first direct in vivo evidence that active PGs production in CL occurs up to Day 4 after estrus in the cow.

Areolae of the Placenta in the Antarctic Minke Whale (<i>Balaenoptera bonaerensis</i>)

Journal of Reproduction and Development, 2014

In this study, we examined the existence and structure of areolae and the steroidogenesis of areo... more In this study, we examined the existence and structure of areolae and the steroidogenesis of areolar trophoblast cells in the Antarctic minke whale placenta morphologically and immunohistochemically. Placentas were collected from the 15th, 16th and 18th Japanese Whale Research Program under Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. The opening and cavity of fetal areolae formed by taller columnar trophoblast cells (areolar trophoblast cells) with long microvilli and a bright cytoplasm, as compared with the trophoblast cells of the chorionic villi interdigitating with the endometrial crypts, were recognized in observations of serial sections. The opening of the areolar cavity was hidden by chorionic villi with areolar trophoblast cells. Furthermore, a closed pouch-like structure lined by tall columnar cells similar to areolar trophoblast cells within the stroma of chorionic villi was noticed and continued to the areolar cavity, with the opening seen on serial sections. In a surface investigation of the chorion and endometrium by SEM, maternal (endometrial) areolae irregularly surrounded by endometrial folds were obvious. Moreover, we distinguished areolar trophoblast cells with long microvilli attached with many blebs from trophoblast cells. In our immunohistochemical observations, a steroidogenic enzyme, cytochrome P450 side chain cleavage enzyme (P450scc), was detected with strong immunoreactivity in trophoblast cells. However, areolar trophoblast cells showed weak or no immunoreactivity for P450scc.

Hyaluronan regulates sperm-induced inflammatory response by enhancing sperm attachment to bovine endometrial epithelial cells via CD44: in-silico and in-vitro approaches

Frontiers in Endocrinology

Recently, we reported that sperm induce cluster of differentiation 44 (CD44) expression and Toll-... more Recently, we reported that sperm induce cluster of differentiation 44 (CD44) expression and Toll-like receptor 2 (TLR2)-mediated inflammatory response in bovine uterus. In the present study, we hypothesized that the interaction between CD44 of bovine endometrial epithelial cells (BEECs) and hyaluronan (HA) affects sperm attachment and thereby enhancing TLR2-mediated inflammation. To test our hypothesis, at first, in-silico approaches were employed to define the binding affinity of HA for CD44 and TLR2. Further, an in-vitro experiment using the sperm-BEECs co-culture model was applied to investigate the effect of HA on sperm attachment and inflammatory response. Here, low molecular weight (LMW) HA at different concentrations (0, 0.1, 1, or 10 µg/mL) was incubated with BEECs for 2 h followed by the co-culture without- or with non-capacitated washed sperm (106/ml) for additional 3 h was performed. The present in-silico model clarified that CD44 is a high-affinity receptor for HA. Moreo...

Activation of sperm Toll-like receptor 2 induces hyperactivation to enhance the penetration to mucus and uterine glands: a trigger for the uterine inflammatory cascade in cattle

Frontiers in Immunology, Dec 18, 2023

Activation of sperm Toll-like receptor 2 induces hyperactivation to enhance the penetration to mu... more

ヤギ精巣網液および精巣上体液中のインヒビン活性について

Nihon Chikusan Gakkaiho, 1987

Seasonal changes in inhibin activity in seminal plasma and serum concentrations of FSH, LH and testosterone in the male goat ()

Theriogenology, 1987

Inhibin activity in goat seminal plasma was measured by in vitro assay throughout successive 9 mo... more Inhibin activity in goat seminal plasma was measured by in vitro assay throughout successive 9 months and its relationship with the serum FSH, LH and testosterone concentrations was investigated. Total inhibin activity (TIA) in seminal plasma gradually increased from spring to summer, reduced in autumn (P<O.O5) and recovered toward winter (P<O.O5). Serum FSH and LH reached a peak in midsummer (P<O.Ol) and returned to the low levels in autumn. Serum testosterone also increased in midsummer and kept the high levels until the early winter (P<O. 05). Some positive correlation was found in monthly levels between seminal TIA and serum FSH (r=0.305; P<O.O5). Results suggest that the summer increase of inhibin activity in seminal plasma relates with the midsummer rise of serum FSH levels in the male goat.

Evidence that oviduct-sperm-immune cells interaction ensures Th2 environment in bovine oviduct immunity for protecting sperm survival

Zearalenone interferes with the sperm-triggered inflammation in the bovine uterus in vitro: Negative impact on sperm motility and survival

Reproductive Toxicology, 2022

Zearalenone (ZEN)-contaminated diets induce detrimental effects on the bovine reproduction. Recen... more Zearalenone (ZEN)-contaminated diets induce detrimental effects on the bovine reproduction. Recently, we reported that active sperm induce pro-inflammatory responses in bovine endometrial epithelial cells (BEECs) in vitro. This study aimed to investigate the impact of presence of ZEN on the sperm-uterine crosstalk in vitro. BEECs monolayers were stimulated by ZEN (10, 100, and 1000 ng/mL) for 0, 3, 6, 12, or 24 h and gene expressions were analyzed by real-time PCR. Moreover, BEECs were pre-exposed to ZEN (10, 100, and 1000 ng/mL) for 24 h then, co-incubated with sperm for 6 h. Conditioned media (CM) from a sperm-BEECs co-culture, after pre-exposure to ZEN, were harvested and exploited to challenge either polymorphonuclear cells (PMNs) or sperm. Both PMNs phagocytic activity toward sperm and sperm motility parameters were then assessed. Results showed that ZEN alone induced pro-inflammatory responses in BEECs through the induction of mRNA expressions of pro-inflammatory cytokines (TNFA and IL1B) and PGES1 at different time points. Pre-exposure of BEECs to ZEN, amplified the sperm-triggered upregulation of pro-inflammatory cytokines (TNFA and IL1B) and chemokine IL8 mRNA abundance in BEECs. Sperm-BEECs conditioned media, primed by ZEN, stimulated the PMNs phagocytosis for sperm whereas suppressed sperm motility parameters. Taken together, these findings indicate that the presence of ZEN augments the pro-inflammatory cascade triggered by sperm in BEECs, provokes PMNs phagocytosis for sperm, and reduces sperm motility parameters. Such immunological reactions may create a hostile environment for sperm competence and survival in the bovine uterus, thus impair fertility.

Intrauterine infusion of low levels of interferon‐tau on day‐8 post‐estrus stimulates the bovine endometrium to secrete apolipoprotein‐A1: A possible implication for early embryo tolerance

American Journal of Reproductive Immunology, Jul 10, 2022

Toll-like receptor 2 of sperm is involved in sperm attachment to bovine oviductal epithelial cells

Sperm TLR2 regulates Ca2+ influx and acrosome reaction to facilitate sperm penetration to oocytes during in vitro fertilization in cows

An acute-phase protein, alpha 1-acid glycoprotein, is a local regulator to protect sperm from neutrophil phagocytosis in the bovine oviduct

The oviduct is classically described as a sterile milieu, though pathogens and endotoxins could i... more The oviduct is classically described as a sterile milieu, though pathogens and endotoxins could invade its mucosal surfaces via the uterus, peritoneal cavity and follicular fluid. We have recently shown that polymorphonuclear neutrophils PMNs , the first line of defense against microorganisms, are present in the bovine oviduct fluid during preovulatory stages. We further suggested that the bovine oviduct provides a PGE-rich microenvironment to protect sperm from phagocytosis by PMNs that they possibly face in vivo, thereby supporting sperm survival in the oviduct. Alpha 1-acid glycoprotein AGP is a major acute-phase protein produced mainly in the liver, having an immunomodulatory functions. In this study, investigated, 1 the local production of AGP in the bovine oviduct, 2 the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production, and 3 the impact of AGP desialylation on the PMN phagocytosis of sperm. AGP gene was expressed in cultured bovine oviduct epithelial cells BOECs and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps NETs for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to secrete PGE2, and AGP and PGE2 at local levels additively inhibited sperm phagocytosis. The results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.

Evidence that bovine oviduct secrets AGP (a1-acid glycoprotein) to favor a Th2 response and to suppress sperm phagocytosis by neutrophils

Peptidoglycan disrupts early embryo-maternal crosstalk via suppression of ISGs expression induced by interferon-tau in the bovine endometrium

Biochemical and Biophysical Research Communications, Oct 1, 2020

Uterine infection with bacteria and the release of peptidoglycan (PGN), antigenic cell wall compo... more Uterine infection with bacteria and the release of peptidoglycan (PGN), antigenic cell wall components of both Gram-negative and Gram-positive bacteria, can cause early pregnancy losses in ruminants, but the associated mechanisms remain unsolved. Day 7 blastocyst starts to secrete a minute amount of interferon-tau (IFNT) in the uterine horn which is required for early stage of maternal recognition of pregnancy (MRP) in ruminants, and it induces interferon stimulated genes (ISGs) for driving uterine receptivity in cows. This study investigated if PGN disrupts IFNT response through modulation of endometrial ISGs expressions. Cultured bovine endometrial epithelial cells (BEECs) were treated with embryo culture medium (ECM) or IFNT (1 ng/ml) in the presence or absence of a low level of PGN (10 pg/ ml) for 24 h. A real-time PCR analyses revealed that the presence of PGN suppressed IFNT-induced ISGs (OAS1 and ISG15) and STAT1 expressions in BEECs. To visualize the impact of PGN in an ex-vivo model that resembles the in vivo status, endometrial explants were treated by IFNT (1 ng/ml) with or without PGN (10 pg/ml) for 12 h. PGN suppressed IFNT-induced gene expressions of the above factors, but not for IFNA receptor type1 (IFNAR1) or type2 (IFNAR2) in explants. Immunofluorescence analysis illustrated that PGN completely suppressed the IFNT-triggered OAS1 protein expression in the luminal epithelium of explants. Of note, PGN did not stimulate pro-inflammatory cytokines (TNFA and IL1B) or TLR2 mRNA expression in both models. These findings indicate that the presence of low levels of PGN suppresses ISGs expression induced by IFNT secreted from early embryo, at the luminal epithelium of the bovine endometrium. This could severely interfere with early stage of MRP processes in cows, leading to pregnancy failure.

Bovine oviduct epithelial cells suppress the phagocytic activity of neutrophils towards sperm but not for bacteria in vitro: Immunofluorescence and electron microscopic observations

Histology and Histopathology, Oct 17, 2019

Previously, we reported that polymorphonuclear neutrophils (PMNs) are constantly existent in the ... more Previously, we reported that polymorphonuclear neutrophils (PMNs) are constantly existent in the bovine oviduct fluid during the pre-ovulatory stage under physiological conditions. Moreover, incubation of PMNs with bovine oviduct epithelial cells-conditioned medium (BOEC-CM) resulted in suppression of their phagocytic activity for sperm. During pathophysiological conditions, cows may be inseminated by infected semen which exposes oviductal PMNs to allogenic sperm simultaneously with pathogens. This study aimed to visually investigate the role of oviduct epithelium in regulating the phagocytic behavior of PMNs toward sperm as a physiological stimulus, with Escherichia coli (E. coli) as a pathological stimulus. In our experiment, PMNs were incubated for 2 h in BOEC-CM. Phagocytosis was then assayed by co-incubation of these PMNs either with sperm, E. coli, or latex beads. BOEC-CM significantly suppressed the direct phagocytosis of PMNs for sperm, but did not affect their phagocytic activity for E. coli or latex beads. Additionally, an investigation with scanning electron microscopy revealed that BOEC-CM suppressed the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. BOEC-CM did not alter NETs formation towards E. coli. A quantification of NETs formation using an immunofluorescence microscopy showed that the areas of NETs formation for E. coli were significantly larger than those formed for sperm. Our data clearly show that the bovine oviduct, through secretions, protects sperm from phagocytosis by PMNs and eliminates bacterial dissemination through maintaining the phagocytic activity of PMNs towards bacteria.