Hepatic stellate cells (HSCs) play essential roles in normal and pathophysiological function in liver. In steady state, HSCs contribute to retinoid storage, immune tolerance, and extracellular matrix (ECM) homeostasis. Upon liver injury, they become activated and lead to morphological and functional changes. Studies have demonstrated that activation of HSCs by various stimuli such as toxins, microbial infection, or metabolic overload can promote the fibrotic changes in liver by production of ECM. Herein, we provide current knowledge about the basic characteristics of HSCs and the mechanism by which they are activated.
The Dromedary camel (Camelus dromedaries) is an important species because of its ability to produce good quality meat, milk, and fibers under harsh environmental conditions. Camels are also crucial for transportation, racing, and as draft animals in agriculture. Therefore, dromedary camels play a critical role in the economy for millions of people living in the arid part of the world. The inherent capability of camels to produce meat and milk is highly correlated with their reproductive performance. Compared with other domestic species, the reproductive efficiency in camelids is low. Although recent reproductive technologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) have been successfully applied to camelids and the birth of live offspring following these technologies has been reported; in vitro embryo production (IVP) has lagged in this species. The development of the IVP system for dromedary camels may be a useful tool for the genetic improvement of this species. IVP in farm animals includes three main steps; in vitro maturation (IVM) of an oocyte, IVF of a matured oocyte, and in vitro culture (IVC) of fertilized oocyte up to the blastocyst stage. This review aims to summarize various factors that influence oocyte quality, IVM, and in vitro embryo development in dromedary camel.
The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode’s albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.
The aim of the current study was to compare the effects of season and breed on the reproductive performance of male and female sheep using 12 rams and 318 ewes of Assaf and Awassi breeds under the seasonal environmental condition of United Arab Emirates for two years. The blood level of testosterone hormone was measured monthly. Semen was collected twice a month from each male using artificial vagina and evaluated for volume, motility, livability, abnormality and concentration. The scrotal circumference and thickness as well as the left testicular length, width, height and volume were measured at one-month intervals. The level of testosterone in Assaf breed was significantly higher in autumn than winter and summer. The scrotal circumference and thickness as well as the left testicular length were significantly higher in Assaf breed than Awassi breed. While, left testicular width and volume were significantly higher in Awassi breed than Assaf breed. Scrotal circumference which was higher in spring and summer than in autumn and winter season in both breeds. The SCC of semen was significantly higher in autumn than in other seasons in both breeds. The sperm abnormality was significantly higher in summer than other seasons in both breeds. The livability was significantly lower in summer in both breeds. Fecundity and prolificacy were significantly higher in Assaf than Awassi breed during autumn season. Assaf breed showed the superior reproductive performance in the autumn season when compared with Awassi breed in the same season and other seasons. The Assaf breed tolerated the climatic conditions in UAE and kept the litter size of 1.72 in comparison to Awassi breed which showed litter size of 1.09. in conclusion, the results showed the superiority of Assaf over Awassi breed and offer a good model of breeding with increased fecundity and prolificacy specially in autumn season.
Equine follicle stimulating hormone receptor (eFSHR) has a large extracellular domain and an intracellular domain containing approximately 10 phosphorylation sites within the G protein-coupled receptor. This study was conducted to analyze the function of phosphorylation sties at the eFSHR C-terminal region. We constructed a mutant of eFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 641 (eFSHR-t641). This removed 10 potential phosphorylation sites from the C-terminal region of the intracellular loop. The eFSHR-wild type (eFSHR-wt) and eFSHR-t641 cDNAs were subcloned into the pCMV-ARMS1-PK2 expression vector. These plasmids were transfected into PathHunter CHO-K1 Parental cells expressing β-arrestin 2 enzyme acceptor fusion protein and analyzed for agonist-induced cAMP response. The cAMP response in cells expressing eFSHR-t641 was lower than the response in cells expressing eFSHR-wt. EC50 values of eFSHR-wt and eFSHR-t641 were 1079 ng/mL and 1834 ng/mL, respectively. eFSHR-t641 was approximately 0.58-fold compared with that of eFSHR-wt. The maximal response in eFSHR-wt and eFSHR-t641 was 24.7 nM and 16.7 nM, respectively. The Rmax value of phosphorylation sites in eFSHR-t641 was also decreased to approximately 68.4% of that in eFSHR-wt. The collective data implicate that the phosphorylation sites in the eFSHR C-terminal region have a pivotal role in signal transduction in PathHunter CHO-K1 cells, and indicate that β-arrestin is involved in coupling the activated receptors to the internalization system.
Partheno Embryo's research is known to play a very important role in identifying the development of embryonic cells or analyzing the genetic mechanisms of embryonic development, but the information on apoptosis formed during the early stage of development on Partheno Embryo is very little. Therefore, this study analyzed whether the embryonic cell death of unit embryos can be inhibited by adding Scriptaid, one of HDACi, which plays a role in demethylation of histone proteins as a method of regulating the cell cycle in the early embryo development of Partheno Embryo. As a result, the differentiation rate was higher in the group that added Scriptaid and FBS, but the cellular development was higher in the group that added pregnant serum to Scriptaid. As a result of analyzing the expression of the gene through IF and PCR, the group with the addition of gestational serum increased the expression of BCL2 and PCNA, which affects the anti-Casp3 action in cell survival. In addition, it is interpreted that treatment of Scriptaid for 16 hours, rather than 24 h treatment lowers the expression of Casp-3, a representative factor of apoptosis, and also increases embryonic development, thus affecting early embryo development. Therefore, it is concluded that the 16-hour treatment of Scriptaid and the use of gestational serum will inhibit cell death in the early embryonic development and increase the development rate of the embryo.
Advancing the estrous cycle of mares is an essential breeding strategy that is routinely conducted by Thoroughbred breeders to improve economic outcomes. For this purpose, Equilume light masks have been developed as an alternative to existing technologies such as artificial lighting or hormonal treatments because they are considered as valid as existing methods with additional animal welfare advantages. For example, with the Equilume light masks, horses can be let out into the pasture, whereas they have to be kept indoors during lighting treatment. Because the function of Equilume light mask on the estrous cycle of mares is influenced by environmental factors such as nutrition condition and temperature, Equilume light mask should be studied in various environments. The objective of the present study was thus to verify the effect of Equilume light masks on the onset of the estrous cycle in Thoroughbred mares in South Korea. Mares were randomly selected and separated into two groups at two Thoroughbred horse breeding farms. The mares in the treatment group were equipped with Equilume light masks from November 18 to February 10 the following year. The body condition, the number and size (> 35 mm) of uterine follicles, and the uterine horn score of the mares were assessed on January 6 and February 10. The body condition scores were not different between the two groups. The treatment with the Equilume light mask had no positive effects on developing follicles and the reproductive tract. In conclusion, the use of Equilume light masks did not influence the seasonal resumption of the estrus cycles in Thoroughbred mares in South Korea.
The present study aimed to investigate the conceptus-related changes during early pregnancy in the Black Bengal breed of goat. A total of 22 gravid genitalia of the Black Bengal goats were collected from local slaughterhouses. The crown-rump lengths (CRL) of the conceptuses were determined to estimate the gestational age (GA). The length and diameter of uterine horn and amniotic sac were measured, and volume of amniotic and allantoic fluid formed by individual conceptus were recorded. The results reveal that the CRL is positively correlated with GA of the conceptus (R2 = 0.89, p < 0.05); however, CRL was not influenced by number of conceptus. Both the left and the right uterine horn gradually increased in size with the advancement of pregnancy irrespective of conceptus number present in the horn. The size of the amniotic sac of conceptus gradually increased with the conceptus age but maintained spherical shape from 5 to 7 weeks of pregnancy. The amniotic fluid formed by individual conceptus rapidly increased from 5 weeks (3.4 ± 0.3 mL) to 7 weeks (21.0 ± 2.0 mL) and 9 weeks (111.5 ± 4.0 mL). The volume of allantoic fluid formed by individual conceptus was steadily increased until 7 weeks (60.0 ± 5.0 mL) and began to decline slowly thereafter (50.0 ± 5.0 mL at 9 weeks). Notably, there was no effect of conceptus number per pregnancy on individual amniotic and allantoic fluid volume. The cotyledons have first appeared on the allanto-chorionic surface from 4 to 5 weeks of pregnancy. The closed eye, nostril and hooves of the conceptus became visible at 7 weeks of pregnancy. The present study has shown the basic information on conceptus-related developmental changes during early pregnancy up to 9 weeks in Black Bengal goat.
The pregnancy rate in indigenous ewes inseminated with frozen-thawed Suffolk semen following natural and synchronized estrus was determined. The serum Progesterone and Estrogen concentration and vaginal electrical resistance (VER) of ewes at the time of Artificial Insemination (AI) were observed as successful pregnancy determinants. 21 healthy ewes were selected for this experiment during January-April, 2017. 10 ewes were inseminated in natural estrus. Whereas, 11 ewes were inseminated after estrus synchronization using intravaginal sponges containing 60 mg medroxyprogesterone acetate. Trans-cervical Al (TCAI) was performed in all ewes within 12-16 hours of observed heat. Prostaglandin E1 analogue impregnated vaginal sponge was used for cervical relaxation 6-8 hours before insemination. Pregnancy was diagnosed through trans-abdominal ultrasonography after 40 days of AI. The pregnancy rate of ewes in synchronized estrus was higher (54.5%) than in natural estrus (30%). Higher serum Progesterone level (0.90 ± 0.02 ng/mL) and significantly (p < 0.001) lower VER (257.78 ± 10.11 ohm) were observed at the time of AI in ewes becoming pregnant. Results suggest that higher Progesterone concentration and lower VER could be considered as pregnancy indicators. Oestrous synchronization could be implemented to increase the pregnancy rate in ewes.
The effects of the number of frozen-thawed ram sperm per single and double intra-cervical artificial insemination (AI) on fertility in ewes were studied. A total of 89 non-pregnant ewes were synchronized for oestrus with two doses of 100 μg PGF2α (Cloprostenol) 9 days apart. The ewes were randomly assigned to one of four groups; D200 (n = 23; double AI with 200 × 106 sperm), S200 (n = 24; single AI with 200 × 106 sperm), D100 (n = 24; double AI with 100 × 106 sperm) and S100 (n = 18; single AI with 100 × 106 sperm). Ewes were inseminated within 12 to 18 h for single AI and, within 10 to 12 h and 16 to 18 h for double AI after the onset of oestrus. The onset of oestrus ranged from 28 to 76 h (54.33 ± 1.28 h). The high percentage (29.2%) of ewes showed oestrus between 51 to 60 h. The non-return rates were highest in group D200 (56.5%) and differed significantly (p < 0.05) from group S100 (11.1%). No ewes were pregnant in group S100, and the pregnancy rates among the remaining groups did not differ. The mean gestation period was 152.8 ± 0.5 days and no difference was observed among the groups. The lambing and multiple birth rates were 100% in group D200. The single and twin lambing was highest in group D100 (33.3%) and group D200 (83.3%), respectively. Only one triplet lambing and the highest lambing size (2.2 ± 0.2) was recorded in group D200. In conclusion, double AI with 200 × 106 sperm showed comparatively most practical for achieving high pregnancy rates and lambing performances in Bangladeshi ewes under field conditions.
Food and agricultural production sector, especially livestock production is vital for Mongolia’s economic and social development. Domestic sheep play key roles for Mongolians, providing food (meat, milk) and raw materials (wool, sheepskin), but genetic diversity, origin of sheep populations in Mongolia have not been well studied. Studies of population genetic diversity is important research field in conservation and restoration of animal breeds and genetic resources. Therefore, this study aimed to investigate genetic characteristics and estimate origin through the analysis of mitochondrial DNA control region D-loop and Cytochrome b of Mongolian indigenous sheep (Mongolian native, Orkhon and Altanbulag) and one Europe sheep (Suffolk). As a result of there were found, 220 SNPs (Single nucleotide polymorphism) in the D-loop region, 28 SNPs in the Cytochrome B region, furthermore, 77 Haplotypes. The nucleotide diversity was only found in D-loop region (n = 0.0184). Phylogenetic analysis showed that 3 (A, B, and C) of 5 haplogroups of sheep have been identified in our research. Haplogroup C was only found in Mongolian indigenous sheep. Haplogroup D and E were not observed. As a result of haplogroups, haplogroup A was dominant (n = 46 of 94 sheeps), followed by haplogroup B (n = 36) and haplogroup C (n = 12). Sequence analysis showed that T deletion, insertion and heteroplasmy in D-loop region occurred at a high rate in Mongolian indigenous sheep population (T insertion = 47, T deletion = 83). The heteroplasmy, which has never been found in Mongolian sheep, has been newly discovered in this study. As a result, the Mongolian sheep varieties, which mainly derived from Asia, were in hybridization with European sheep varieties.
Cryopreservation is used for blastocyst preservation of most mammalian embryos and is an important technique for breeding. We aimed to compare the efficiency of the cryopreservation method using the standard Cryotop device and the ReproCarrier device, a domestic product manufactured in Korea. The efficacy of the two devices was analyzed based on the survival rate, intracellular levels of reactive oxygen species (ROS), and apoptosis of the vitrified bovine blastocysts. The survival rates of the vitrified-warmed blastocysts were similar between the ReproCarrier group (58.4 ± 17.7%) and Cryotop group (59.9 ± 14.1%). Intracellular ROS levels and apoptotic index were determined by DCFDA staining and TUNEL assay. Changes in intracellular ROS levels, number of total nuclei, and cellular apoptosis of vitrified blastocysts after cryopreservation were not significantly different between the two groups. These results indicate that the ReproCarrier device method is as effective as the standard Cryotop method for vitrification of bovine blastocysts in vitro.