Evidence for existence of insulin-like factor 3 (INSL3) hormone-receptor system in the ovarian corpus luteum and extra-ovarian reproductive organs during pregnancy in goats.

Insulin-like factor 3 (INSL3), initially described as a male hormone, is expressed in female reproductive organs during the estrous cycle and pregnancy but its function has not yet been established. This study explores the function of INSL3 in pregnant Saanen goats by characterizing the expression dynamics of INSL3 and its receptor, relaxin family peptide receptor 2 (RXFP2) and by demonstrating specific INSL3 binding in reproductive organs, using molecular and immunological approaches and ligand-receptor interaction assays. We demonstrate that the corpus luteum (CL) acts as both a source and target of INSL3 in pregnant goats, while extra-ovarian reproductive organs serve as additional INSL3 targets.

Effects of pre-freeze Nigella sativa oil supplementation on cryosurvival of ovine spermatozoa.

The study was designed with three experiments to evaluate the effects of pre-freeze supplementation of Nigella sativa oil (NSO) and thymoquinone (TQ) on total motility, progressive motility, biokinetic characteristics, acrosomal integrity and DNA integrity of cryopreserved ovine spermatozoa. Semen samples collected from three proven fertile Merino rams were diluted with a Tris-based cryomedia containing different levels of NSO (Experiment I: 0, 10, 100 and 1,000 g/ml), TQ (Experiment II: 0, 1, 10 and 20 g/ml) and their optimum levels (Experiment III: 100 g/ml of NSO, 10 g/ml of TQ and 1 mM of α-tocopherol and cryopreserved as pellet (200µL) and subsequently evaluated at different post-thaw incubation periods (0, 2 and 4 hr). The results revealed that the percentage of total motility, progressive motility and biokinetic characteristics such as average path velocity, curvilinear velocity and straight-line velocity were higher (p < 0.

Possible ability of bovine follicular fluid to attract migrating bull spermatozoa.

Aim: To examine the potential of bovine follicular fluid (BFF) to attract bull spermatozoa.

Methods: The ability of the BFF to attract bull sperm was evaluated by observing changes in sperm migration after being placed in a cross-column chamber. The movement parameters of the heads and flagella of the sperm that were attracted to the BFF were analyzed by using the Computer Assisted Sperm Analysis system.

Involvement of calcium channels and intracellular calcium in bull sperm thermotaxis.

Thermotaxis that sperm migrate to higher temperature area has been confirmed in rabbit and human. In this study, we examined the migration ability of bull sperm in a temperature gradient to confirm thermotaxis and elucidate the involvement of calcium in such thermotaxis, as well as the relation between sperm capacitation and bull fertility. Thermotaxis was evaluated in a temperature gradient of 34-42ºC using a cross-type column 22-mm long, 40-mm wide, and 100-μm deep.

Involvement of Transient Receptor Potential Vanilloid (TRPV) 4 in mouse sperm thermotaxis.

Transient Receptor Potential Vanilloid (TRPV) 4 is one of the temperature-sensitive ion channels involved in temperature receptors, and it is known to be activated from 35 to 40ºC. Here we analyzed sperm motility function of Trpv4 knockout (KO) mouse in temperature-gradient conditions to elucidate the thermotaxis of mouse sperm and the involvement of TRPV4 in thermotaxis. The sperm were introduced at the vertical column end of a T-shaped chamber filled with medium in a plastic dish, and we measured the number of sperm that arrived at both ends of the wide column where we had established a temperature gradient of approx.

Expression of insulin-like factor 3 hormone-receptor system in the reproductive organs of male goats.

Relaxin-like factor (RLF), generally known as insulin-like factor 3 (INSL3), is essential for testis descent during fetal development. However, its role in adult males is not fully understood. We investigate the function of INSL3 in male Saanen goats by identifying cell types expressing its receptor, relaxin/insulin-like family peptide receptor (RXFP)2 and by characterizing the developmental expression pattern of INSL3 and RXFP2 and the binding of INSL3 to target cells in the male reproductive system.

The active form of goat insulin-like peptide 3 (INSL3) is a single-chain structure comprising three domains B-C-A, constitutively expressed and secreted by testicular Leydig cells.

Relaxin-like factor (RLF), also called insulin-like peptide 3 (INSL3), is a member of the insulin/relaxin gene family and is produced by testicular Leydig cells. While the understanding of its effects is growing, very little is known about the structural and functional properties of native INSL3. Here, we demonstrate that native INSL3 isolated from goat testes is a single-chain structure with full biological activity, and is constitutively expressed and secreted by Leydig cells.

The active form of goat insulin-like peptide 3 (INSL3) is a single-chain structure comprising three domains B-C-A, constitutively expressed and secreted by testicular Leydig cells.

Abstract Relaxin-like factor (RLF), also called insulin-like peptide 3 (INSL3), is a member of the insulin/relaxin gene family and is produced by testicular Leydig cells. While the understanding of its effects is accumulating, very little is known about the structural and functional properties of native INSL3. Here, we demonstrate that native INSL3 isolated from goat testes is a single-chain structure with full biological activity, and is constitutively expressed and secreted by Leydig cells.

Partial cDNA sequence of a relaxin-like factor (RLF) receptor, LGR8 and possible existence of the RLF ligand-receptor system in goat testes.

Relaxin-like factor (RLF), also known as insulin-like factor 3 (INSL3), is produced by testicular Leydig cells, but its specific receptor LGR8 (leucine-rich repeat family of G-protein-coupled receptor 8) has not been identified in goats. This study aimed to identify complementary DNA (cDNA) sequences of goat LGR8, and characterize the expression of both RLF and LGR8 in goat testes by RT-PCR and immunohistochemistry. Testes were collected from immature (3-month-old) and mature (24-month-old) Saanen goats, and partial cDNA sequences of the goat homologue of human LGR8 were identified.

Effect of selenium and vitamin E on acrosome reaction in porcine spermatozoa.

Purpose: Selenium (Se) and vitamin E (Vit-E), as an integral part of antioxidant systems, play an important role in the motility and acrosome reaction (AR) of mammalian spermatozoa. The purpose of this study was to investigate the effect of Se and Vit-E on motility, viability, AR and accumulation of ammonia in the culture medium during different incubation periods in porcine sperm.

Methods: Sperm samples were washed, swum-up and incubated at 37°C for 1 and 3 h in Sp-TALP medium supplemented with sodium selenite (SS), seleon l-methionine (SeMet) and Vit-E in the presence or absence of ammonia.

Metabolism of exogenous fatty acids, fatty acid-mediated cholesterol efflux, PKA and PKC pathways in boar sperm acrosome reaction.

Purpose: For understanding the roles of fatty acids on the induction of acrosome reaction which occurs under association of cholesterol efflux and PKA or PKC pathways in boar spermatozoa, metabolic fate of alone and combined radiolabeled C-oleic acid and H-linoleic acid incorporated in the sperm was compared, and behavior of cholesterol and effects of PKA and PKC inhibitors upon fatty acid-induced acrosome reaction were examined.

Methods: Semen was collected from a Duroc boar, and the metabolic activities of fatty acids in the spermatozoa were measured using radioactive compounds and thin layer chromatography. Cholesterol efflux was measured with a cholesterol determination assay kit.

Effect of amino acids and dipeptides on the acrosome reaction and accumulation of ammonia in porcine spermatozoa.

The present study was designed to investigate the effect of amino acids and their dipeptides in the medium related to the urea cycle on the motility, viability, acrosome reaction (AR) and accumulation of ammonia in the medium over different incubation periods in porcine spermatozoa and to assess the utilization of glucose. Porcine spermatozoa were washed, swim-up and incubated at 37°C for 0-4 h in mTALP medium supplemented with 75-600 µmol/L ammonia. Amino acids (1.

Sex preselection in bovine by separation of X- and Y-chromosome bearing spermatozoa.

Flow cytometrically-sorted sperm has been involved in the production of sex preselected offspring. More than 30,000 bovine offspring have been produced using AI and other means using spermatozoa separated by flow cytometer. Flow cytometric sperm sorting based on differences in their DNA content is the best method for separation of X- and Y-chromosome bearing spermatozoa.

Development of Mongolian gerbil embryos in chemically defined media: effects of osmolarity, glucose and phosphate.

Mongolian gerbil 2-cell embryos were cultured in modified M16. When osmolarity of the medium with 5.0 mmol glucose l(-1) was varied by adjusting the amount of NaCl added, 2-cell embryos at 280, 290, 300 and 310 mOsmol developed to the 8- and 16-cell stages.