Methionine promotes the development of hair follicles via the Wnt/β-catenin signalling pathway in Rex rabbits

To investigate the effect and molecular mechanism of methionine (Met) on the growth of hair follicles (HFs) in Rex rabbits. A total of 200 weaning Rex rabbits were divided into four groups and fed varying levels of Met-supplemented diets. We measured the HF density on dorsal skin and the Wnt/β-catenin pathway protein expression level. Meanwhile, whole HFs were isolated from Rex rabbit skins and cultured with Met in vitro to measure hair shaft growth. The relationship between Met and the Wnt/β-catenin signalling pathway was also characterized by using the Wnt/β-catenin signalling inhibitor, XAV-939. The results showed that the addition of dietary Met could significantly increase the HF density on dorsal skin (p < .05) and enhance the protein expression level of Wnt10b (p < .05), β-catenin (p < .05) and DSH (p < .05). Methionine stimulation could also prolong the hair shafts growth in vitro (p < .05). And inhibition of Wnt/β-catenin signalling using XAV-939 could eliminate this phenomenon. In summary, Met can increase the density of HFs on dorsal skin in vitro and prolong the hair shaft growth of HFs in vivo via the Wnt/β-catenin signalling pathway.     

母体埋植褪黑激素对子一代獭兔内脏组织中RORα基因表达的影响

本研究旨在探讨獭兔母体埋植褪黑激素(MT)对RORα基因在子一代獭兔内脏组织中的表达及影响,选择110只相近月龄的健康母獭兔同日配种,第5日后选出已确定妊娠的母兔76只,随机平均分为2组,对照组母兔不做处理,试验组母兔埋植10 mg MT。在仔兔出生第1天时每组选15只宰杀,取皮肤、肌肉、肺脏、心脏、肝脏、肾脏及十二指肠组织样品,测定仔兔组织样品中RORα基因的表达量,结果表明:RORα基因在子一代獭兔各组织中的表达量与表达模式均不同,该基因在子一代獭兔皮肤组织中的表达水平极显著上调,在肌肉、肺脏组织中显著上调,在肝脏组织中极显著下调,在心脏、肾脏组织中显著下调,在十二指肠中的表达与对照组相比无显著变化。     

Polymorphism of the Melatonin Receptor Genes and its Relationship with Seasonal Reproduction in the Gulin Ma Goat Breed

Melatonin is thought to be the main molecule that transmits the signal of seasonal change to the neuroendocrine system in seasonal breeding species. Melatonin exerts its effects through specific melatonin receptors, MTNR1A and MTNR1B. In the present study, six native goat breeds in China and one introduced goat breed were analysed to investigate the relationship between the genetic polymorphism of receptor genes and seasonal reproduction. Sequencing results showed that there were five polymorphic mutations in the MTNR1A gene and two in the MTNR1B gene. In the MTNR1A gene, genotypes AA, AB and BB for 424C>T and genotypes CC, CD and DD for 589C>A were observed in these goat breeds. In all six native goat breeds, only genotype AA was detected. In the MTNR1B gene, genotypes EE, EF and FF for 1179G>A and genotypes GG, GH and HH for 1529A>G were detected. However, in Gulin Ma goats, the genotypes EE and HH were not found. Moreover, the base of G at position 1179 and A at position 1529 were linked (By Arlequin ver 3.1, Zoological Institute, Berne, Switzerland, http://cmpg.unibe.ch/software/arlequin3 ,D′ = 0.7496, r² = 0.4421, χ² = 489.8679, p = 0.000). Among these mutations, no amino acid change was found in MTNR1A, while both of the mutations in MTNR1B gene caused amino acid changes of R222H and S339G, respectively. The structural analysis showed that the R222H mutation occurred in the first amino acid residue of the third cytoplasmic loop, and the S339G mutation was located in the carboxyl terminus of the protein. In terms of seasonal breeding, all the genotypes we detected showed a similar kidding frequency distribution trend with a higher frequency in May–August than in January–April and in September–December. This suggests that the relationship between the polymorphisms in the MTNR1A and MTNR1B genes and seasonal breeding could not be established.     

Polymorphisms of the melatonin receptor 1A gene that affects the reproductive seasonality and litter size in Small Tail Han sheep

Previous researches have shown that MTNR1A plays an essential role in sheep reproduction. However, most researches focused more on the reproductive seasonality of sheep, and few scientists had studied the association of polymorphisms of the MTNR1A gene with ovine litter size and reproductive seasonality. Therefore, we chose MTNR1A gene to detect its novel sequence polymorphisms and population genetics and analyse their association with seasonal reproduction and litter size in ewes. The mRNA expression level in hypothalamus, pituitary and ovary was also detected. In this study, five polymorphisms (g.15118664G > T, g.15118683C > T, g.15118756C > T, g.15118774C > T and g.15118951G > A) were identified in exon 2. Most importantly, the g.15118683C > T and g.15118951G > A were significant difference between year‐round oestrous sheep and seasonal oestrous sheep (p < .01), and g.15118756C > T had a great effect on litter size of Small Tail Han sheep (p < .05). In addition, the mRNA expression level of MTNR1A in the hypothalamus of polytocous Small Tail Han sheep was significantly higher than that in monotocous Small Tail Han sheep (p < .01) and the expression of MTNR1A in the hypothalamus of year‐round oestrous sheep was significantly higher than that in seasonal oestrous sheep (p < .01). Polymorphisms in exon 2 may regulate the reproductive seasonality and litter size of ewes by influencing gene expression to regulate the reproductive seasonality and litter size of ewes. Our studies provided useful guidance in marker‐assisted selection of the litter size in Small Tail Han sheep.     

Effect of melatonin on regulation of apoptosis and steroidogenesis in cultured buffalo granulosa cells

This study was aimed to address melatonin receptor expression, mRNA level of hypothalamus and hypophysis hormone receptors (GnRHR, FSHR, and LHR), steroidogenesis, cell cycle, apoptosis, and their regulatory factors after addition of melatonin for 24 hr in cultured buffalo granulosa cells (GCs). The results revealed that direct addition of different concentrations of melatonin (100 pM, 1 nM, and 100 nM) resulted in significant upregulation (p < 0.05) of mRNA level of melatonin receptor 1a (MT1) without affecting melatonin receptor 1b (MT2). Melatonin treatment significantly downregulated (p < 0.05) mRNA level of FSH and GnRH receptors, whereas 100 nM dose of melatonin significantly increased mRNA level of LH receptor. Treatment with 100 nM of melatonin significantly decreased the basal progesterone production with significant decrease (p < 0.05) in mRNA levels of StAR and p450ssc, and lower mRNA level of genes (Insig1, Lipe, and Scrab1) that affect cholesterol availability. Melatonin supplementation suppressed apoptosis (100 nM, p < 0.05) and enhanced G2/M phase (1 nM, 100 nM, p < 0.05) of cell cycle progression which was further corroborated by decrease in protein expression of caspase‐3, p21, and p27 and increase in bcl2. Our results demonstrate that melatonin regulates gonadotrophin receptors and ovarian steroidogenesis through MT1. Furthermore, the notion of its incorporation in apoptosis and proliferation of buffalo GCs extends its role in buffalo ovaries.     

Expression of melatonin and its related synthase and membrane receptors in the oestrous corpus luteum and corpus luteum verum of sheep

Melatonin is an important factor involved in regulating reproduction; it is synthesized enzymatically by the sequential action of melatonin‐synthesizing enzymes, arylalkylamine N‐acetyltransferase (AANAT) and hydroxyindole‐O‐methyltransferase (HIOMT), and exerts its biological functions mainly through receptor‐mediated action. To evaluate the expression of melatonin, two melatonin‐synthesizing enzymes (HIOMT and AANAT), and membrane receptors (MT1 and MT2) in oestrous corpus luteum (CL) and CL verum of sheep (Ovis aries), we performed ELISA, qRT‐PCR, western blotting and immunohistochemistry. The quantitative results showed that melatonin, HIOMT and AANAT levels in the CL verum were significantly higher than those in oestrous CL (p < 0.05), whereas MT1 and MT2 exhibited no change between the oestrous CL and CL verum (p > 0.05); moreover, the localization results showed that HIOMT, AANAT, MT1 and MT2 were mainly expressed in large luteal cells (LLCs). In summary, the above results suggested that sheep CL has potential for the synthesis of melatonin; meanwhile, they also suggested that CL is one of the targets of melatonin. These results provide not only a basis for whether sheep CL can synthesize melatonin but also provide a reference for further study on the mechanism of melatonin in the CL.     

褪黑激素对獭兔日增重及其毛皮皮尺的影响

选取体重为1.4kg左右日龄相近的健康獭兔60只,随机分为3组,每组20只,公母各半。分别皮下埋植褪黑激素0mg、10mg、20mg,研究褪黑激素对獭兔日增重及其毛皮皮尺的影响。结果表明:褪黑激素对獭兔的日增重增加4.1%(P>0.05)、毛皮长度以及毛皮厚度均无显著(P>0.05)影响;但埋植10mg组的獭兔毛皮宽度要显著(P<0.05)大于埋植20mg组。     

饲粮中谷氨酰胺添加水平对断奶至3月龄獭兔毛皮品质和肠道屏障的影响

本试验旨在研究饲粮中谷氨酰胺添加水平对断奶至3月龄獭兔毛皮品质和肠道屏障的影响。选用体重相近的断奶獭兔180只,随机分为5组,每组36个重复,每个重复1只兔。各组分别饲喂谷氨酰胺添加水平为0(对照)、0.3%、0.6%、0.9%和1.2%的试验饲粮。预试期7 d,正试期56 d。结果表明:饲粮中谷氨酰胺添加水平对生长獭兔毛皮面积、毛皮重量、被毛长度和被毛厚度无显著影响(P0.05)。饲粮中谷氨酰胺添加水平对生长獭兔十二指肠绒毛高度/隐窝深度值有显著影响(P0.05)。与对照组相比,饲粮中添加0.9%谷氨酰胺显著升高了空肠中闭合小环蛋白mRNA的表达量(P0.05),显著降低了空肠中丙酮酸激酶mRNA的表达量(P0.05);此外,饲粮中添加0.9%谷氨酰胺显著增加了十二指肠黏膜中分泌性免疫球蛋白A含量(P0.05)。综上所述,饲粮中谷氨酰胺添加水平没有影响到生长獭兔的毛皮品质,但改善了肠道机械屏障和免疫屏障功能。在本试验条件下,断奶至3月龄獭兔饲粮中谷氨酰胺适宜的添加水平为0.9%。     

Immunolocalization of Melatonin and Follicle‐Stimulating Hormone Receptors in Caprine Ovaries and their Effects During in vitro Development of Isolated Pre‐Antral Follicles

The expression of melatonin type 1 (MT1) and FSH (FSHR) receptors in caprine ovaries and the effects of these hormones on the in vitro development of isolated pre‐antral follicles were evaluated. Follicles (≤200 μm) were cultured for 12 days in α‐MEM (control) or melatonin (100 or 1000 pg/ml) or sequential melatonin medium (100 pg/ml: from day 0 to day 6; 1000 pg/ml: from day 6 to day 12; experiment 1) and in control or sequential FSH (100 ng/ml from day 0 to day 6; 500 ng/ml from day 6 to day 12) or sequential melatonin or this latter plus sequential FSH (experiment 2). MT1 and FSHR expressions were observed in granulosa cells from secondary and antral follicles. The oocytes from primordial and primary follicles also express FSHR. Sequential melatonin increased the percentage of normal follicles and oocyte recovery compared with the control or melatonin (1000 pg/ml) at day 12. In experiment 2, all the treatments increased the normal follicles and growth compared with the control. In conclusion, this study demonstrated the presence of MT1 and FSHR in caprine ovaries. The addition of increased concentrations of melatonin (sequential medium) or FSH can be used to promote the in vitro development of caprine pre‐antral follicles.     

Melatonin improves cryopreservation of ram sperm by inhibiting mitochondrial permeability transition pore opening

Cryopreservation damages permeability of sperm mitochondrial membranes, with formation of a mitochondrial permeability transition pore (mPTP). Mitochondria are both a primary synthesis site and principle target for melatonin, which can directly inhibit mPTP formation. The objective was to determine effects of melatonin on mPTP opening of frozen-thawed ram sperm and elucidate underlying pathways by antagonist and agonists of melatonin receptors (MTs), and antagonists of PI3K and GSK 3β treatments; furthermore, plasma membrane integrity, mitochondrial membrane potential (ΔΨm), mitochondrial cytochrome c (Cyt c) release and fertilization were analysed to assess the effect of mPTP status mediated by melatonin on quality of frozen-thawed sperm. Fresh ram semen was diluted in glucose-egg yolk buffer with 0 or 10^–7 M melatonin (frozen and frozen + melatonin groups, respectively) and slow-frozen. In frozen-thawed sperm, melatonin added at initiation of 4°C equilibration was most effective for inhibiting mPTP opening, decreasing peptidyl-prolyl-cis/trans isomerase activity of cyclophilin D and increasing plasma membrane integrity, ΔΨm, mitochondrial Cyt c concentration and fertilizing ability (p < .05). In a mechanistic study, the melatonin receptor (MT)1 antagonist eliminated inhibition of melatonin on mPTP opening, whereas MT1 agonist had opposite effects (p < .05). Neither MT2 antagonist nor agonist had significant effect, but PI3K and/or GSK 3β antagonist decreased inhibition of MT1 agonist on mPTP opening (p < .05). In conclusion, melatonin improved sperm cryopreservation, perhaps by acting on MT1 via the PI3K-Akt-GSK 3β pathway to inhibit mPTP opening.     

褪黑激素对獭兔生长性能及皮毛质量的影响

本试验旨在研究褪黑激素对獭兔生长性能及皮毛质量的影响。采用单因子试验设计,选取55~60日龄体重相近的健康生长獭兔200只,随机分为4组,每组50只兔。对照组饲喂基础饲粮,试验组分别饲喂含不同水平褪黑激素(10、25、40 mg/kg)的试验饲粮。试验期为99 d。结果表明:60~150日龄,各试验组平均日增重均极显著高于对照组(P0.01),各试验组料重比均极显著低于对照组(P0.01)。90、120、130、150日龄各试验组被毛密度均极显著高于对照组(P0.01)。40 mg/kg试验组臀部、肩部、腹部的皮张厚度均显著或极显著低于对照组(P0.05或P0.01)。由此可见,褪黑激素能够提高獭兔平均日增重,降低料重比,同时提高獭兔被毛密度,促进獭兔皮张提前成熟。     

Pyridoxine regulates hair follicle development via the PI3K/Akt,Wnt and Notch signalling pathways in rex rabbits

This study was conducted to evaluate the effect of pyridoxine on the development of hair follicles in Rex rabbits and the underlying molecular mechanism. Two hundred 3-month-old Rex rabbits were randomly divided into 5 groups and fed diets supplemented with 0, 5, 10, 20, or 40 mg/kg pyridoxine. The hair follicle density on the dorsal skin and the gene and protein expression levels of components of the phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB or Akt), Wnt, Notch and bone morphogenetic protein (BMP) signalling pathways were measured. In addition, free hair follicles were isolated from Rex rabbits and cultured with pyridoxine in vitro to measure hair shaft growth. Furthermore, dermal papilla cells (DPC) were isolated from the skin of Rex rabbits and cultured with pyridoxine in vitro to measure the gene and protein expression levels of components of the PI3K/Akt, Wnt, Notch and BMP signalling pathways. The results showed that the addition of dietary pyridoxine significantly increased the total follicle density, secondary follicle density, and secondary-to-primary ratio (S/P, P < 0.05), that the growth ratio of hair stems was promoted by pyridoxine in basic culture medium, and that the growth length of tentacle hair follicles cultured in the pyridoxine group was longer than that in the control group (P < 0.05). In addition, pyridoxine changed the DPC cycle progression and promoted cell proliferation, and appropriate concentrations of pyridoxine (10 and 20 μmol/L) significantly inhibited cell apoptosis (P < 0.05). Pyridoxine significantly affected the gene expression of components of the PI3K/Akt, Wnt and Notch signalling pathways in the skin and DPC of Rex rabbits (P < 0.05), increased the levels of phosphorylated catenin beta 1 (CTNNB1) and Akt, and decreased the level of phosphorylated glycogen synthase kinase 3 beta (GSK-3β) (P < 0.05). Therefore, the molecular mechanism by which pyridoxine promotes hair follicle density in Rex rabbits probably occurs through activation of the PI3K/Akt, Wnt and Notch signalling pathways, prolonging hair follicle growth and delaying the onset of telogen.     

Effect of melatonin on visceral fat deposition,lipid metabolism and hepatic lipo-metabolic gene expression in male rats

Melatonin (MT) influences lipid metabolism in animals; however, the mechanistic effect of melatonin on liver fat and abdominal adipose deposition requires further clarity. In order to study the effects of melatonin on lipid metabolism, and hepatic fat and abdominal adipose deposition in animals, twenty Sprague–Dawley (SD) rats of 6 weeks of age with similar bodyweight were randomly divided into two groups: control (CTL) and MT-treated (10 mg/kg/day). During a 60-day experiment, food intake and bodyweight were measured daily and weekly respectively. At the end of treatment, blood samples were collected to collect plasma to quantify hormones and metabolic indicators of lipid metabolism. In addition, organ and abdominal adipose depots including liver, and omental, perirenal, and epididymal fat were weighed. Liver tissue was sampled for sectioning, long-chain fatty acid (LCFA) quantification, and gene chip and Real-time quantitative PCR (qPCR) analyses. The results showed that liver weight and index (ratio of liver weight to body weight) in MT group reduced by 20.69% and 9.63% respectively; omentum weight and index reduced by 59.88% and 54.93% respectively, and epididymal fat weight reduced by 45.34% (p = 0.049), relative to CTL. Plasma lipid indices, triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL) and total cholesterol (TC) with MT treatment decreased significantly compared with the control. Fat and 8 LCFA content in liver in MT group also decreased. Gene chip and qPCR demonstrated that there were 289 genes up-regulated and 293 genes down-regulated by MT. Further analysis found that the mRNA expression of lipolysis-related genes increased, while the mRNA expression of lipogenesis-related enzymes decreased (p < 0.05) with MT. This study concluded that melatonin greatly affected fat deposition, and hepatic LCFA supply and the expression of genes associated with lipogenesis and lipolysis.     

Influence of multi-enzyme preparation supplemented with sodium butyrate on growth performance blood profiles and economic benefit of growing rabbits

The present study was carried out to explore the impacts of dietary supplementation of enzyme mixture with sodium butyrate on the growth performance, carcass traits, blood profile and economic benefit in two breeds of weanling rabbits adapted to survive in Egypt (New Zealand White and Rex). One-hundred and twenty weaned male rabbits (New Zealand White and Rex) of 6 weeks of age and 770.5 ± 20 g body weight were allotted randomly into four groups in a factorial arrangement. The obtained results indicated that there were non-significant differences in all growth performance traits, blood profile and economic parameters due to the breed effect. However, there were significant differences in most of carcass traits due to the breed effect except total giblets and New Zealand White breed showed the highest value of these parameters including dressing % (p < .01), forequarter and loin % (p < .001) and hindquarter % (p < .003) compared with Rex breed counterparts. The effect of the treatment and its interaction with the breed significantly (p < .05) improved body weight gain, feed consumption and carcass traits (percentage of dressing, forequarter, hind quarter and lion). However, final body weight and feed conversion ratio were not significantly influenced. Supplementing a diet with treatment significantly decreased blood triglycerides, cholesterol and the ratio between albumin and globulin (A/G ratio), while increased blood total protein and globulin. Although higher feed cost and total costs in treated groups than control ones in each breed, they showed higher total return and net return. Rex non-treated rabbit breed showed the lowest profitability measures compared with other groups. In conclusion, dietary supplementation of multi-enzyme with sodium butyrate is highly recommended in growing rabbits due to their beneficial effects on the growth performance and profitability.     

The Effect of Lactobacillus isolates on growth performance,immune response,intestinal bacterial community composition of growing Rex Rabbits

The objective of this experiment was to investigate the effect of Lactobacillus isolates on intestinal bacterial community composition of growing Rex Rabbits. A total of 120 weaned Rex Rabbits (30 days old, 30 per group) were used for the experiment, which started after an adaptation period of 7 days. The control group was fed with basal diet only, while the treatment I, II and III groups were fed with basal diet adding antibiotics, Lactobacillus zeae (LB1) and Lactobacillus casei (L3) respectively. Growth performance, immune response and intestinal flora have been examined. The results obtained were as follows: (i) F/G of the rabbits fed with Lactobacillus isolates was significantly lower than that of the control group (p < 0.05). (ii) The concentration of ALT decreased significantly (p < 0.05) and that of IgG and IgM increased significantly (p < 0.05) after feeding rabbits with Lactobacillus isolates. (iii) Lactobacillus isolates had no influence on the number of mast cells in duodenum and jejunum, but increased the number of mast cells in caecum significantly (p < 0.05). (iv) The data from pyrosequencing‐based analysis suggested that the bacterial community in the rabbit's intestinal flora can be changed by Lactobacillus isolates and antibiotics, especially for the microbial diversity and abundance in the caecum. In conclusion, the application of proper Lactobacillus isolates can improve the growth performance, enhance the immunological function and adjust the intestinal micro‐ecosystem of growing Rex Rabbits.     

Inhibition of prolactin promotes secondary skin follicle activation in cashmere goats

The aim of this study was to investigate the involvement of prolactin (PRL) on development of secondary skin follicles in cashmere goats. Goats were randomly assigned to either a bromocriptine treatment or control group. Samples of cashmere fiber, blood, and skin were collected from all goats after 1 mo. The results indicated that the length, growth rate, and diameter of fibers were not influenced (P > 0.05) by the inhibition of PRL resulting from the treatment with bromocriptine. There was a tendency for increases in total follicle number, primary and secondary follicle numbers, and in the ratio of secondary to primary follicles following treatment with bromocriptine, but these differences were not significant (P > 0.05). The percentage of active secondary follicles in anagen was increased (P < 0.05) in the bromocriptine-treated goats, but there was no effect of treatment on the percentage of active primary follicles. Bromocriptine decreased (P < 0.05) circulating concentrations of PRL and Insulin-like growth factor 1 (IGF1) and increased (P < 0.05) those of melatonin (MT), but there was no effect of this treatment on the serum concentrations of cortisol, growth hormone, tetraiodothyronine, and triiodothyronine. In bromocriptine-treated goats, mRNA expressions of PRL and MT membrane receptor 1a (MTNR1a) were decreased (P < 0.05) and mRNA expression of MT nuclear receptor (RORα) was increased (P < 0.05), but there was no effect of the treatment on expression of long PRL receptor, short PRL receptor, MT membrane receptor 1b and IGF1. It is concluded that inhibition of PRL promotes secondary hair follicle development in the anagen phase, possibly by downregulating MTNR1a and up-regulating RORα gene expression in the skin.     

Growth of foetal bones and metabolic profile during gestation in primiparous ewes and multiparous ewes

Primiparous ewes and multiparous ewes show physiological differences during pregnancy, which can have an impact on the development of their offspring. The objective of this study was to compare the changes in the metabolic profile and in the size of some foetal bones throughout gestation between primiparous and multiparous ewes. Twelve primiparous (PM) ewes and 14 multiparous (MT) ewes were used. According to the dates of lambing, two groups of ewes were formed: Group 1 (G1, n = 6 PM and n = 7 MT) and Group 2 (G2, n = 6 PM and n = 7 MT). The body weight, body condition score, metabolic and foetal morphometric parameters were determined from before conception until the end of gestation. After lambing, the body weight and survival rate during the first 72 hr of life of lambs, as well as the maternal behaviour score were recorded. The PM ewes were lighter (p < .01) and had a greater mobilization of body reserves during gestation, mainly evidenced by a greater serum concentration of NEFAs and lower serum concentration of total proteins (p < .05) compared with the MT ewes. The parity did not affect the foetal morphometric variables. The lambs of MT ewes were heavier at parturition (p = .002) and tended to have a greater survival rate than those lambs of PM ewes (p = .09). In conclusion, PM ewes and MT ewes differ in their metabolic profile throughout the gestation. However, in the present study, we did not find parity differences in the dimensions of foetal bones during growth in gestation.     

The expression pattern,polymorphisms and association analyses of the porcine NREP gene

NREP (neuronal regeneration related protein homolog) plays a role in the transformation of neural, muscle, and fibroblast cells and in smooth muscle myogenesis. The NREP gene was selected for detailed study as an expressional and functional candidate gene on the basis of data from the expression microarray, which detected the differences in gene expression between Czech Large White pigs and wild boars in the longissimus lumborum et thoracis and biceps femoris muscle tissues. Quantitative real-time PCR results confirmed that porcine NREP was expressed in both skeletal muscles and significantly overexpressed in Czech Large White pigs compared with wild boars (14.5- and 11.6-fold; p < .05). We identified 9 polymorphic sites in the genomic DNA of NREP. Six of these polymorphisms were in complete linkage disequilibrium, and therefore, only 4 loci were informative. The associations of the HF571253:g.103G>A, HF571253:g.134G>A, HF571253:g.179T>C and HF571253:g.402_409delT polymorphisms with backfat thickness, lean meat content and average daily gain were assessed in Czech Large White pigs. The GG genotypes HF571253:g.103G>A and HF571253:g.134G>A, the TT genotypes HF571253:g.179T>C and 67 HF571253:g.402_409delT genotypes had favourable effects on the studied traits. Our results indicate the possibility of utilizing the variability of the NREP gene in marker-assisted selection in order to improve meat production in pigs.     

Presence of melatonin‐catabolizing non‐specific enzymes myeloperoxidase and indoleamine 2,3‐dioxygenase in the ram reproductive tract

The melatonin catabolism is very complex and not completely understood. Melatonin can be metabolized by free radical interaction, but also pseudo‐enzymatically or by enzymatic pathways. We have previously detected the existence of melatonin‐synthesizing enzymes and melatonin receptors MT1 and MT2 in the ram reproductive tract; thus, in order to start to elucidate melatonin catabolism in these organs, we have investigated the presence of the melatonin‐catabolizing enzymes indoleamine 2,3‐dioxygenase (IDO, both IDO1 and IDO2 isoforms) and myeloperoxidase (MPO) in testis, epididymis and accessory glands. Gene expression analyses by real‐time PCR showed the presence of MPO, IDO1 and IDO2 in all the organs of the ram reproductive tract and revealed that MPO is the main melatonin‐catabolizing enzyme, which is mainly expressed in the testis and the bulbourethral glands (p < .05). These results were further corroborated by immunohistochemical staining, and by Western blot. Likewise, MPO was also evidenced in epididymal and ejaculated spermatozoa by indirect immunofluorescence and Western blot. In conclusion, melatonin‐catabolizing enzymes MPO, IDO1 and IDO2 are expressed in the ram reproductive tract, and MPO is the most expressed one, mainly in the testis and the bulbourethral glands. The presented results warrant further studies on the function of these enzymes and their melatonin‐metabolizing activity.