Altered luteal expression patterns of genomic and non-genomic progesterone receptors in bitches at different reproductive states.

The binding of steroid hormones to their specific receptors is necessary to exert their effects on target cells. Progesterone (P4), a steroid hormone, carries out its effects through both genomic and non-genomic (the cell membrane-associated) receptors. This study aimed to ascertain luteal expression patterns of genomic and non-genomic progesterone receptors in bitches in physiological (early dioestrus and early pregnant) and pathological (pyometra) reproductive states. Luteal tissue was collected from the bitches at early dioestrus (ED, n = 5), early pregnant (EP, n = 5), and pyometra (PY, n = 5). The expression profiles of Steroidogenic Acute Regulator Protein (STAR), Progesterone Receptor (PGR), Membrane Progestin Receptors (PAQR5, PAQR7 and PAQR8), and Progesterone Membrane Components (PGMRC1 and PGMRC2) were examined at the mRNA levels using Real-Time Polymerase Chain Reaction (RT-PCR). Protein levels of PGR, PGMRC1 and PGMRC2 were detected by western blotting (WB). The STAR expression was found in all groups, with a statistical difference observed between EP and PY groups (P < 0.05). The protein level of PGR was determined to be highest in the EP group and lowest in the PY group. The expression of PAQR8 increased in the EP group (P < 0.05). The PAQR5 exhibited high expression in the EP group and low expression in the PY group (P < 0.05). PGRMC1 was more elevated in the EP group and lower in the PY group (P < 0.05). Protein levels of PGMRC1 and PGMRC2 were also observed at the highest expression in EP group. According to the altered expression profiles for examined receptors, we suggest that those progesterone receptors have roles in early pregnancy or pyometra in bitches.

An investigation of the distributions of ferroptosis and necroptosis mediators in the maternal-fetal interface at different days of rat pregnancy.

Ferroptosis and necroptosis are recognized as playing major roles in the regulation of various physiological processes. However, the physiological role of the cell death mediated by these two pathways in the developmental process has not yet been clearly established. This study investigated ferroptosis and necroptosis signalling pathways in maternal-fetal tissue in the different gestational days (GD) of rat pregnancy using immunohistochemical and western blot methods in order to fill this gap. Twenty-four female Wistar albino rats were mated and divided into three groups. Maternal-fetal tissue samples were collected on GD 5, 12 and 19 of pregnancy. Expression and total protein levels of the markers glutathione peroxidase-4, soluble transporter family 7 member 11, transferrin receptor, receptor-interacting serine/threonine-protein kinase 1, receptor-interacting serine/threonine-protein kinase 3 and mixed lineage kinase domain-like protein were investigated on both the maternal and fetal surfaces of the placenta using immunohistochemical and western blot methods. The results showed varying levels of protein expression of both ferroptosis and necroptosis mediators in the GD 5, 12 and 19 of pregnancy. Immunohistochemical analyses revealed that these mediators were located on both the maternal (decidua and metrial gland) and fetal surfaces (labyrinth zone, yolk sac and basal zone) and that their expression levels changed in the different GD. The findings revealed the existence of important ferroptosis and necroptosis pathway mediators in rat maternal-fetal tissue. These results may provide a molecular framework for a better understanding of the communication between the placenta, decidua and fetus during the developmental process.

Oleuropein Mitigates Acrylamide-Induced Nephrotoxicity by Affecting Placental Growth Factor Immunoactivity in the Rat Kidney.

OBJECTIVE: Oleuropein is one of the main components of the antioxidant properties of olive leaves. Placental growth factor is an important regulator in angiogenesis and inflammation, its levels being variable in pathological conditions. In this study, we aimed to examine changes in placental growth factor expression and the effect of oleuropein, found in olive leaves, in rats exposed to acrylamide nephrotoxicity. MATERIAL AND METHODS: Twenty-four male Wistar albino rats were allocated into 4 groups. The control group received saline solution only. The oleuropein group received oleuropein (4.2 mg/kg), the acrylamide group received acrylamide (5 mg/kg), and the acrylamide and oleuropein group received acrylamide (5 mg/kg) and oleuropein (4.2 mg/kg). All substances were administered via gastric gavage for 21 days. Kidney tissues were removed at the end of the study and subjected to histopathological, stereological, and immunohistochemical procedures. RESULTS: Histopathological examination revealed dilatation, vacuolization, and degeneration in the proximal and distal tubules and increased placental growth factor immunoreactivity in the acrylamide group. Cavalieri volume analysis revealed increased cortex, distal, and proximal tubule volumes (P < .01). CONCLUSION: Oleuropein significantly attenuated acrylamide-induced kidney injury by altering placental growth factor immunoreactivity. Placental growth factor immunoreactivity can be used as a marker of acrylamide nephrotoxicity, and oleuropein may counteract acrylamide-induced kidney injury.

An investigation of the endoplasmic reticulum stress in obesity exposure in the prenatal period.

OBJECTIVES: Exposure to maternal obesity has been shown to make offspring more prone to cognitive and metabolic disorders later in life. Although the underlying mechanisms are unclear, the role of endoplasmic reticulum (ER) stress in the fetal programming process is remarkable. ER stress can be activated by many chronic diseases, including obesity and diabetes. Therefore, our study aimed to investigate the role of ER stress caused by maternal diet-induced obesity in the offspring hippocampus. We also evaluated the protective effect of N-acetylcysteine (NAC) against ER stress. METHODS: A rat obesity model was created by providing a high-fat (60 % kcal) diet. N-acetylcysteine (NAC) was administered at a dosage of 150 mg/kg via the intragastric route. The animals were mated at the age of 12 weeks. The same diet was maintained during pregnancy and lactation. The experiment was terminated on the postnatal 28th day, and the offspring's brain tissues were examined. Immunohistochemical staining for ER stress markers was performed on sections taken from tissues after routine histological procedures. RESULTS: The results revealed increased GRP78, PERK, and eIF2α immunoreactivities in the hippocampal dentate gyrus (DG) and cornu ammonis 1 (CA1) regions in the obese group offspring, while the expression of those markers in those regions normalized with NAC supplementation (p < 0.01). Statistical analysis of XBP1 immunoreactivity H-scores revealed no difference between the study groups (p > 0.05). DISCUSSION: These results suggest that exposure to obesity during the prenatal period may cause increased ER stress in hippocampal neurons, which have an important role in the regulation of learning, memory and behavior, and this may contribute to decreased cognitive performance. On the other hand, NAC stands out as an effective agent that can counteract hippocampal ER stress.

Expression of placental growth factor and a disintegrin and metalloprotease with a thrombospondin type motifs 1-4-8 during the three trimesters of rat pregnancy at the maternal-fetal interface.

The functional roles of the a disintegrin and metalloprotease with a thrombospondin type motifs (ADAMTS) gene family in reproductive physiology, reproductive organs developments and adult reproductive health are still under investigation. The expression of the anti-angiogenic proteases ADAMTS-1, ADAMTS-4 and ADAMTS-8 in placental angiogenesis at various stages of pregnancy also remains unclear. The purpose of this study was therefore to determine the localization and expression of the ADAMTS-1, ADAMTS-4 and ADAMTS-8 proteins during the three stages of pregnancy in rats. Maternal-fetal tissue samples were collected on Days 5, 12 and 19 of each trimester, corresponding to the first, second and third trimesters. The expression of placental growth factor (PlGF) and ADAMTS-1, ADAMTS-4 and ADAMTS-8 at the maternal-fetal interface was examined using immunohistochemistry and western blot at three distinct phases of pregnancy. ADAMTS-1, ADAMTS-4 and ADAMTS-8 were detected in all three trimesters of pregnancy. The relative amount of PIGF increased in the first trimester and decreased significantly in the third trimester (p < 0.05). The expression of ADAMTS-1 and ADAMTS-4 was significantly higher in the second (p < 0.05) and third trimesters (p < 0.01) compared to the first trimester. However, no statistically significant change was observed in ADAMTS-8 expression between trimesters. The ADAMTS exhibiting the highest expression during the first trimester was ADAMTS8. These findings indicate that the expression of ADAMTS-1, ADAMTS-4 and ADAMTS-8 in the three different stages of rat pregnancy may be involved in the modulation of decidualization, morphogenesis and angiogenesis. Periodic changes in ADAMTS expression are thought to be regulated by gonadal steroids.

Expression of ADAMTS 1-4-8 and placental growth factor in ovary and oviduct during pregnancy in the first trimester.

A Disintegrin and Metalloprotease Domains with Thrombospondins Motifs (ADAMTS), proteinases responsible for the destruction of extracellular matrix structures, have essential roles in the physiological and pathological processes of the female reproductive system, which is a dynamic structure. This study aimed to evaluate the immunoreactivity of placental growth factor (PLGF) and ADAMTS' (1, -4, and -8) in the ovary and oviduct during pregnancy in the first trimester. Our findings suggest a predominant role of ADAMTS-4 and ADAMTS-8 as proteoglycan-degrading enzymes from the ADAMTS-1 in the first trimester. As an angiogenic factor, PLGF showed more immunoreactivity than ADAMTS-1 in the ovary. This study provides the first evidence that ADAMTS-4 and ADAMTS-8 are more expressed in ovarian cells and follicles at different developmental stages during the first trimester of pregnancy than ADAMTS-1. Consequently, we suggest that ADAMTSs and PLGF act together and may exert specific effects on the formation, stabilisation, and function (or a combination thereof) of the matrix surrounding and protecting the follicles.

Protective and therapeutic role of melatonin against tunicamycin-induced ER stress in testicular tissue of rats.

Objectives: This study aimed to investigate the possible consequences of administering exogenous melatonin as prevention or treatment against tunicamycin-induced endoplasmic reticulum (ER) stress in the testicular tissue of rats. Materials and Methods: In this study, 42 adult Sprague Dawley rats, randomly divided into seven equal groups, were administered intraperitoneal tunicamycin to induce ER stress. Both prophylactic (PMel) and therapeutic melatonin (TMel) groups were administered melatonin for seven days. ER stress in the cell was detected through immunohistochemical and molecular analyses using GPR78 expression. Results: Increased oxidant levels and apoptosis rates were shown in testicular tissue because of ER stress. The sections in the melatonin-administered and control groups were similar, with melatonin-administered groups showing an increase in the antioxidant ratio. Histometric examinations revealed both TMel and melatonin applications reduced the diameter of the tubules. However, immunohistochemical and molecular analyses showed that PMel administration decreased the concentration of GRP78 more effectively than TMel. Conclusion: Applying melatonin prior to cell damage occurrence can be recommended for its effectiveness in protecting from tunicamycin-induced ER stress.

The mixture of liquid foam soap, ethanol and citric acid as a new fixative-preservative solution in veterinary anatomy.

The present study investigates the efficiency of liquid foam soap, ethanol, citric acid and benzalkonium chloride as a fixative-preservative solution (a soap-and ethanol-based fixing solution, or SEFS). In this study, ethanol serves as the fixative and preservative, liquid foam soap as the modifying agent, citric acid as the antioxidant and benzalkonium chloride as the disinfectant. The goat cadavers perfused with SEFS (n=8) were evaluated over a period of one year with respect to hardness, colour and odour using objective methods. Colour and hardness were compared between one fresh cadaver and the SEFS-embalmed cadavers. Histological and microbiological examinations were also performed in tissue samples. Additionally, the cadavers were subjectively evaluated after dissection and palpation. The SEFS provided the effectiveness expected over a 1-year embalming period for the animal cadavers. No bacteria or fungi were isolated except for some non-pathogenic Bacillus species. Visible mould was not present on either cadavers or in the surrounding environment. The cadavers maintained an appearance close to their original anatomical appearance, with muscles having good hardness and elasticity for dissection.