Recent Advances in Research on the Hormone INSL3 in Male Goats

Insulin‐like factor 3 (INSL3), previously called relaxin‐like factor (RLF), is essential for testis descent during fetal development and has been implicated in the testicular and sperm functions in adult males. However, similar functions in ruminants remain largely unknown. This chapter will cover recent advancement in our understanding of INSL3 in goats. First, testicular Leydig cells were the sole source of INSL3, with INSL3 expression increasing during development. Second, INSL3 was constitutively secreted as a B–C–A single‐chain structure with full biological activity. Third, secreted INSL3 was transported into the seminiferous compartments, where its receptor RXFP2 was expressed on germ cells, thus suggesting that the intratesticular INSL3 hormone‐receptor system operates in germ cells. Fourth, functional RXFP2 enabling INSL3 to bind was also identified in the spermatozoa and suggested the existence of the extratesticular INSL3 hormone‐receptor system in the spermatozoa. Interestingly, percentages of INSL3‐binding spermatozoa were significantly reduced in the semen of subfertile bulls compared to that of fertile bulls, suggesting the potential of this system to diagnose fertility in breeding sires. These fascinating findings will give a new perspective in physiological and/or therapeutic actions of INSL3 on male reproductive processes in domestic ruminants, including goats

Vitrification of Germinal Vesicle Stage Oocytes

In order to cryopreserve germinal vesicle (GV) stage oocytes, we first need to develop a novel container for keeping large quantities of GV oocytes, because of collecting them as cumulus oocytes complexes (COCs) that have bigger size and larger volume than oocytes themselves, and second modify a protocol for optimizing vitrification of them. In this mini-review, we describe our recent progress for attaining these objectives. When 65 bovine COCs having GV oocytes could be placed on a sheet of nylon mesh, and plunged directly into liquid nitrogen for vitrification, the recovery rate was significantly higher compared with that in 15 ones on the electron microscope (EM) grid as a control, followed by obtaining the resultant cleavage and developmental rates after in vitro fertilization and culture (IVFC) without significant difference. Using bovine and murine oocytes, we found that a step-wise manner to expose them with the vitrification solution increased rates of in vitro maturation, subsequent development to blastocysts and hatching/hatched blastocysts after IVFC. Our results show that nylon mesh is an alternative material for cryopreserving large quantities of bovine GV oocytes, and that a step-wise exposure to cryoprotectants may have befit for decreasing disadvantage during vitrification

Gene expression profiles of novel caprine placental prolactin-related proteins similar to bovine placental prolactin-related proteins

BACKGROUND: This study reports the identification of a full-length cDNA sequence for two novel caprine prolactin-related proteins (cPRP1 and cPRP6), and their localization and quantitative expression in the placenta. Caprine PRPs are compared with known bovine PRPs. We examined their evolution and role in the ruminant placenta. RESULTS: Full-length cPRP1 and cPRP6 cDNA were cloned with a 717- and 720- nucleotide open-reading frame corresponding to proteins of 238 and 239 amino acids. The cPRP1 predicted amino acid sequence shares a 72% homology with bovine PRP1 (bPRP1). The cPRP6 predicted amino acid sequence shares a 74% homology with bovine PRP6 (bPRP6). The two cPRPs as well as bPRPs were detected only in the placentome by RT-PCR. Analysis by in situ hybridization revealed the presence of both cPRPs mRNA in the trophoblast binucleate cells. These mRNA were quantified by real-time RT-PCR analysis of the placentome at 30, 50, 90 and 140 days of pregnancy. Both new cPRP genes were able to translate a mature protein in a mammalian cell-expression system. Western blotting established the molecular sizes of 33 kDa for cPRP1 with FLAG-tag and 45 kDa for cPRP6 with FLAG-tag. The sequence properties and localized expression of cPRP1 and cPRP6 were similar to those of bovine. However, their expression profiles differed from those in bovine placenta. Although this study demonstrated possible roles of PRPs in caprine placenta, PRPs may regulate binucleate-cell functions like those in bovine, but their crucial roles are still unclear. CONCLUSION: We have identified the novel PRPs in caprine placenta. Localization and quantitative expression of caprine PRPs were compared with bovine PRPs. The data indicate that PRP genes in caprine placenta have coordination functions for gestation, as they do in bovine. This is the first study of PRPs function in caprine placenta

A Case of a Combination of Wearing-Off-Like Symptoms and Autonomic Hyperreflexia Following Spinal Anesthesia for a Patient with Parkinson’s Disease Who is Treated by Deep Brain Stimulation

Deep brain stimulator (DBS) has been widely performed for various medically refractory movement disorders. We report a 70’s male patients with Parkinson’s disease (PD) treated by DBS in addition to anti-parkinsonian drugs who underwent transurethral lithotomy under spinal anesthesia twice. We canceled his usual medications for PD, therefore, his blood pressure was higher than usual. His DBS was turned off in the operation room before induction of anesthesia. Both anesthesia techniques for TUL surgeries were completed by spinal anesthesia used by 0.5% hyperbaric bupivacine. Immediately after spinal anesthesia in both surgeries, severe rigidity, airway obstruction and conscious change emerged. These complications were suspected wearing-off-likes symptoms of PD, which were proved because of spinal anesthesia itself and/or DBS off effects. Autonomic hyperreflexia with severe hypertension, abnormal sweating and excessive oral secretion was gradually appeared after infusion of normal saline to obtain surgical fields of view, however, these symptoms disappeared after the end of infusion. It was suspected that the adverse events were due to low effects of spinal anesthesia on autonomic nervous system degenerated by Parkinson’s disease. We turned on DBS immediately after the completion of surgeries, and resumed his oral medications on the next day of surgeries. There were no events in his courses. As there is little information and no standard anesthetic guidelines available on patients with DBS implant who are present for surgery, a careful management is needed to avoid complications.本稿の要旨は日本麻酔科学会　中国・四国支部第55回学術集会（松山市，2018年）で発表した

Expression and localization of RLF/ INSL3 receptor RXFP2 in boar testes

This study investigated the possibility of the presence of specific receptor for relaxin-like factor (RLF)/insulin-like peptide 3 (INSL3) in boar testes. While RLF/INSL3 was produced by Leydig cells in the boar testis, its own receptor RXFP2 was expressed mainly in meiotic and post-meiotic germ cells, but not in Leydig cells, suggesting the existence of RLF/INSL3–RXFP2 signaling in germ cells of boars