Pit-1 Gene Research Articles

Transcriptome profiling of four candidate milk genes in milk and tissue samples of temperate and tropical cattle

The expression of four genes involved in milk regulation and production in bovine milk and tissue samples profiled using quantitative PCR to identify differential gene expression. Our goal focussed on the differential mRNA expression of milk genes (KCN, PRL, BLG and PIT-1) in milk samples and different tissues from four different breeds of ecologically adapted and geographically separated cattle species. The mRNA expression identified the four milk genes under studied most upregulated in mammary gland and milk samples as compared with other tissues. The expression of PIT-1 gene in the brain identified to have influenced the expression of PRL and K-CN in the mammary and milk samples. Among the four genes, PRL had the highest mRNA expression (144.19-fold change) in Holstein followed by K-CN with 100.89-fold change, while the smallest relative expression for most genes in this study are in the range from 0.79 to 7.35-fold difference. White Fulani cattle was identified to have a higher expression for K-CN, PRL and BLG compared with Angus and Ndama cattle, while Holstein cattle is on top of the list on the basis of the gene expression and gene regulation for all the four genes in this study. Also, White Fulani and Holstein are in the same cluster based on their mRNA expression for milk genes. Our data showed the first evidence of the molecular identification of indigenous White Fulani cattle ofhaving potential for higher milk production.

Genetic polymorphism of Pit-1|HinfI gene in Grati-Ongole Grade cattle at Indonesian Beef Cattle Research Station

The pituitary transcription factor (Pit-1) gene is one of the considered genes that responsible to growth in cattle. A specific Pit-1|HinfI gene polymorphism located within exon 6(g.1256G>A) has been shown to be associated with growth and milk traits in cattle. This study aimed to identify the Pit-1|Hinf1 gene polymorphism in Grati-Ongole Grade cattle (Grati-OG cattle). A total of 107 genomic DNA of Grati-OG cattle were used in this study. The detection of polymorphism was performed by PCR-RFLP method. Only AB and BB genotypes were succesfully identified with the frequency of 0.009 and 0.991, respectively. Whereas, the frequency of A and B alleles were 0.005 and 0.995, respectively. The observed genotype frequencies in this population do not deviated from HWE. The value of Ho, He and PIC were 0.009, 0.009 and 0.009, respectively.In conclusion, the Pit-1|HinfI gene polymorphism in Grati-OG cattle is monomorphic and hence it should not be used in further association studies.

Short Communication: Association of GH, IGF1R, and PIT1 genes polymorphism with average daily gain and body measurement in Pesisir cattle

Arnim, Yurnalis, Afriani T, Putra DE. 2017. Short Communication: Association of GH, IGF1R, and PIT1 genes polymorphismwith average daily gain and body measurement in Pesisir cattle. Nusantara Bioscience 10: 221-225. Growth hormone (GH) andinsuLin-like growth factor I receptor (IGF1R), and Pit1 gene has important effect on growth, carcass, and meat quality traits in manyspecies. The objectives of the present study were to study the associations of the GH/AluI, GH/MboII, IGF1R/MspI and PIT1/HinfI withgrowth and body measurent such as body length (BL), chest girth (CG), height withers (HW) in Pesisir cattle. A total 175 Pesisir cattlewere used in this study. Weight gain, and body measurement were fitted using the General Linear Model (GLM) procedure of the SASprogram. The resuLts showed significant associations between the CC genotype of the GH/MboII polymorphism and higher weight gainand height withers (p < 0.05). The GH/AluI polymorphism did not have any effect on the weight gain and body measurement. Allelefrequencies were found as A: 0.374 and B: 0.626 in GH/MboII polymorphism in GH gene. The effect of GH/AluI polymorphisms wasnot observed on average daily gain and body measurement in Pesisir cattle (P>0.05). The effect of IGF1R/MSpI polymorphism was notobserved on average daily gain but show significant association between AA and AB genotype and height withers (p < 0.005). ThePIT1/HinfI polymorphism was not associated with average daily gain and body measurement in Pesisir cattle. The resuLts demostratedthat the GH/MboII and GHR1/MspI polymorphisms couLd be used as a candidate gene for selection in Pesisir cattle.

Association of polymorphisms in Pit-1 gene with growth and feed efficiency in meat-type chickens.

ObjectiveThe pituitary specific transcription factor-1 (Pit-1) gene is responsible for pituitary development and growth hormone expression and is regarded as a pivotal candidate gene for growth and production in chickens. Therefore, the aim of this study was to investigate the association of Pit-1 polymorphisms with growth and feed efficiency traits in yellow meat-type chickens.MethodsIn the present study, five single nucleotide polymorphisms (SNPs) of Pit-1 were selected and genotyped by high-throughput matrix-assisted laser desorption-ionization time-of-flight mass spectrometry in 724 meat-type chickens.ResultsAssociation analysis showed that rs13687126 of Pit-1 was strongly associated with body weight gain (BWG) and feed intake (FI) (p<0.05), and that rs13687128 was significantly correlated with body weight at 70 days of age (BW70), BWG and feed conversion ratio (FCR) (p<0.05). SNP rs13905622 was strongly related to BW70 and FCR (p<0.05). Furthermore, birds with the GG genotype of rs13687126 had larger BWG and FI than those with the AG genotype (p<0.05). Individuals with the TT genotype of rs13687128 were significantly higher BW70 and BWG than those of the CT and CC genotype, while FCR was just the opposite (p<0.05). For rs13905622, the AA chickens showed strongly larger BW70 and lower FCR compared with the AT and TT chickens (p<0.05). Additionally, an ACA haplotype based on rs13687126, rs13687128, and rs13905622 had significant effects on BW70 and FCR (p<0.05).ConclusionOur studies thus provide crucial evidence for the relationship between polymorphisms of Pit-1 and growth and feed efficiency traits which may be useful for meat-type chicken breeding programs.

Polymorphism of PIT-1 and Prolactin Genes and Their Effects on Milk Yield in Holstein Frisian Dairy Cows Bred in Vietnam

The aim of this study is to investigate the polymorphism of the Pituitary transcription factor (PIT-1) and Prolactin (PRL) genes in Holstein Friesian cows (HF) bred in Vietnam and their effects on milk yields. For these purposes, 125 HF dairy cows of milking age from Da Lat Milk Farm located in Lam Dong, Vietnam were sampled. Phenotypic data of milk yields (calculated on 305-day of milking cycle; kg/day) of each individual milking cow were collected. The PIT-1.E2, PIT-1.E6, PRL.E3 and PRL.E4 genotypes were analyzed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method with restriction enzymes TaqI, HinfI, RsaI, respectively. The significant effects of PIT-1.E6, PRL.E3 and PRL.E4 polymorphisms on milk yields were not observed in HF dairy cows (P > 0.05). However, results revealed statistically significant associations between PIT-1.E2 polymorphism and milk yields of HF dairy cows; individuals carrying the AA genotype yielded more milk compared to others. Our study suggests the potential use of PIT-1.E2 SNP in marker assisted selection (MAS) for improved milk yields of HF dairy cows bred in Vietnam.

Identification of Pit-1 Gen Using PCR-RFLP of Padjadjaran Sheep and Evaluate of Growth Rate

The objectives of this research were to evaluate variation of Pit-1 gen of Padjadjaran Sheep and evaluate of their growth rate. The data comprised of 15 lambs female and 15 lambs male records of 0 - 12 mouths old lambs. Variation of Padjadjaran Sheep PIT-1 gen was analyzed using PCR-RFLP and used primer by 5’-GAGGGATAATTACAAATGGTCC-3’ and 5’-TGTTAACAGCTGTGGGACACAC-3’, length fragment analyse using HinfI restriction enzyme. Presence or absence of deference bands in a PCR fragment of the result can be distinguished by differences in the electrophoretic migration of the fragment. The result showed that the variation Pit1 gene showed that there was in the position 345 bp, 137 bp and 115 bp. In this experiment the difference migration did occur in all samples (monomorphic), that was amplificated with reverse and forward primer. The average male birth weight (BW), weaning weight (WN) and Yearling Weight (YW) was 2.29 ±0.42kg; 8.96 ±1,89 kg and 30.12 ±5,65 kg. The average female birth weight (BW), weaning weight (WN) and Yearling Weight (YW) was 2.33 ±0.49 kg; 8.23 ±1.99 kg and 26.11 ±5.50 kg. Correlation value between age and body weight was 0,99 for female and 0,97 for male. The highest body weight gain per month was 2.85 kg for female at the age of six months and 3.4 kg for male at the age of one year. The best equition of growth rate was Ŷ = 1,862 + 0,127X1 0,076X2

Relationship of PIT-1 Gene Polymorphism with Breeding Parameters and Body Weights of Cows and Calves

Relationship of PIT-1 Gene Polymorphism with Breeding Parameters and Body Weights of Cows and Calves

The relationships between performance traits and the bGH/Alu I and Pit-1/Hinf I polymorphisms in Holstein cows

Progress in genetic selection in livestock can be increased by marker asisted selection. The identification of favorable genetic markers is one of the most important stages in marker-asisted selection. In this study, it was aimed to determine the effects of the bGH/AluI and Pit-1/HinfI polymorphisms on the production traits of organic reared cows. Genotyping was performed on total 245 Holstein cows, n=181 for Pit-1 gene and n=186 for bGH gene. Milk yields and some reproduction traits analyzed by analysis of variance using the general linear model procedure, and 703 production records from cows were used to. The results showed that neither the Pit-1/Hinf I nor bGH/Alu I polymorphisms affect the tested milk traits (p&gt;0.05).

Identification of Pit1 Gene Variants in Sahiwal Cattle of Pakistan

Identification of Pit1 Gene Variants in Sahiwal Cattle of Pakistan

Allele Fixation in Prolactin and Pit-1 Genes Associated with Highest Fat Content than Milk Yield of Murrah Buffalo Population

Allele Fixation in Prolactin and Pit-1 Genes Associated with Highest Fat Content than Milk Yield of Murrah Buffalo Population

Semen Characteristics and Genotyping of Pituitary-Specific Transcription Factor Gene in Buffalo Using PCR-RFLP

THIS STUDY aimed to evaluate the sperm characteristics in buffalo bulls and to screen the genetic polymorphisms in PIT-1 gene as bases for selection of bulls with good breeding value. The study was performed on 60 buffalo bulls aged 2-8 years. The animals were divided into three groups according to the age. The first group were between 2 to <3 years (n= 35). The second group were between 3 to <5 years (n=14). The third group were between 5 to <8 years (n=11).Three semen collections were obtained from each animal at 15-day intervals and evaluated for volume, individual motility, live sperm and chromatin integrity %. The semen samples were stored at 20 °C until DNA extraction then polymerase chain reaction (PCR) and DNA amplification were carried out.Restriction fragment length polymorphism (RFLP) was used for genotyping of pituitary-specific transcription factor gene using HinfI-RFLP. The results showed that, ejaculate volume, individual motility and live sperm % were significantly lower in old than adult and young buffalo bulls. While chromatin damage percentage had no significant difference among groups. All buffalo bulls were genotyped as BB with the predominance of B allele where PCR 451 bp fragment was digested into two fragments 244 and 207 bp. It may be concluded that age had adverse effect on semen quality. Monomorphic pattern of the amplicon 451 bp in PIT-1HinfI locus in exon 6 was fixed in Egyptian buffalo with the predominance of B allele and BB genotype in a high frequency (100%).

Associations of Leptin and Pituitary-Specific Transcription Factor Genes' Polymorphisms with Reproduction and Production Traits in Dairy Buffalo.

This study aimed to detect the genetic variability in Leptin and Pit-1 genes using polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing also to explore their possible associations with reproductive and productive traits of Egyptian buffaloes. Regarding Leptin gene, three genotypes (AA, AG and GG) were identified with frequency of 0.54, 0.40 and 0.06, respectively, and the genotypes were distributed according to the Hardy-Weinberg equilibrium. Allele A was comparatively higher than G with frequency of 0.74 and 0.26, respectively. For Pit-1 gene, the association could not be performed due to the monomorphism (BB). The results showed that AA genotypes were found to be superior in most of production and reproduction traits. AA genotypes yielded more milk (2332.34kg, p=0.04) with higher fat% (6.10, p=0.004) and fat yield (155.75kg, p=0.06), reach peak milk production at 42.19days and required 2.19 services for conception in comparison with GG genotypes. Birthweight of animals with AA genotype was lesser than with GG genotype (39.35 and 43.67kg, p=0.02, respectively). The days open is numerically better in AA genotype animals (99.35days), but the difference between the three genotypes was non-significant. The distinct significant associations reported in this study suggested that Leptin is reputable candidate genetic marker, which might be used to enhance animals' genetic potential for milk production in conjunction with reproduction.

1-Methyl-2-undecyl-4(1H)-quinolone, a derivative of quinolone alkaloid evocarpine, attenuates high phosphate-induced calcification of human aortic valve interstitial cells by inhibiting phosphate cotransporter PiT-1

An abnormally high serum phosphate level induces calcific aortic stenosis (CAS), which is characterized by ectopic valve calcification and stenosis of the orifice area. Inhibition of ectopic calcification is a critical function of any internal medical therapy for CAS disease. The aim of the present study was to investigate the inhibitory effects of several derivatives of evocarpine, methanolic extracts from the fruits of Evodia rutaecarpa Bentham (Japanese name: Go-Shu-Yu) on the high phosphate-induced calcification of human aortic valve interstitial cells (HAVICs) obtained from patients with CAS. High phosphate (3.2 mM) concentrations significantly increased the calcification of HAVICs after 7 days of culture. This calcification was completely inhibited in the presence of sodium phosphonoformate (PFA), a selective inhibitor of the type III sodium-dependent phosphate cotransporter (PiT-1). PiT-1 contributes to phosphate uptake, resulting in calcification. 1-Methyl-2-undecyl-4(1H)-quinolone (MUQ; 30–300 nM), but not evocarpine or its derivatives dihydroevocarpine and 1-methyl-2-nonyl-4(1H)-quinolone, inhibited the high phosphate-induced HAVICs calcification in a concentration-dependent manner. Although all of the evocarpine derivatives attenuated alkaline phosphatase activity, only MUQ also decreased PiT-1 gene expression with cellular PiT-1 protein diminution. These results suggest that MUQ mitigated high phosphate-induced HAVICs calcification by inhibiting PiT-1 gene expression.

A Pit-1 Binding Site Adjacent to E-box133 in the Rat PRL Promoter is Necessary for Pulsatile Gene Expression Activity.

Recent evidence reveals that prolactin gene expression (PRL-GE) in mammotropes occurs in pulses, but the molecular process(es) underlying this phenomenon remains unclear. Earlier, we have identified an E-box (E-box133) in the rat PRL promoter that binds several circadian elements and is critical for this dynamic process. Preliminary analysis revealed a Pit-1 binding site (P2) located immediately adjacent to this E-box133 raising the possibility that some type of functional relationship may exist between these two promoter regions. In this study, using serum shocked GH3 cell culture system to synchronize PRL-GE activity, we determined that Pit-1 gene expression occurred in pulses with time phases similar to that for PRL. Interestingly, EMSA analysis not only confirmed Pit-1 binding to the P2 site, but also revealed an interaction with factor(s) binding to the adjacent E-box133 promoter element. Additionally, down-regulation of Pit-1 by siRNA reduced PRL levels during pulse periods. Thus, using multiple evidences, our results demonstrate clearly that the Pit-1 P2 site is necessary for PRL-GE elaboration. Furthermore, the proximity of this critical Pit-1 binding site (P2) and the E-box133 element coupled with the evidences of a site-to-site protein interactions suggest that the process of PRL-GE pulse activity might involve more dynamic and intricate cross-talks between promoter elements that may span some, or all, of the proximal region of the PRL promoter in driving its pulsatile expression.

Bovine lactotroph cultures for the study of prolactin synthesis functions.

The aim of this study was to establish a bovine anterior pituitary-derived lactotroph (BAPDL) line that expresses prolactin (PRL) in vitro to study the mechanisms of bovine PRL synthesis and secretion. Immunohistochemistry assay of PRL in the newborn calves' anterior pituitary glands showed that most lactotrophs were located within the superior border of the lateral wings of the anterior pituitary. Tissues of the superior border of the lateral wings of the anterior pituitary were dispersed and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). The limiting dilution method was used to establish BAPDL from single cell clone. BAPDL cells constantly expressed mRNAs for PRL and pituitary-specific transcription factor 1 (Pit-1) gene and grew steadily and rapidly in the DMEM supplemented with 10% FBS. PRL immunoreactivity was present in BAPDL at passage 20. The concentration of bovine PRL in BAPDL at passage 20 culture supernatant was decreased to below 35% compared with that in BAPDL at passage 1. The effects of human epidermal growth factor (hEGF) and dopamine (DA) on the expression and secretion of PRL in BAPDL at passage 4 were also investigated. The results are consistent with those of previous studies. Thus, it can be used successfully for studying the mechanisms of stimuli regulating PRL synthesis and release.

Inhibition of osteoblast mineralization by phosphorylated phage-derived apatite-specific peptide

Functionalization of biomaterials with material- and cell-specific peptide sequences allows for better control of their surface properties and communication with the surrounding environment. Using a combinatorial phage display approach, we previously identified the peptide VTKHLNQISQSY (VTK) with specific affinity to biomimetic apatite. Phosphorylation of the serine residues of the peptide (pVTK) caused a significant increase in binding to apatite, as well as a dose-dependent inhibition of osteoblast mineralization. In this study, we investigated the mechanisms behind pVTK mediated inhibition of mineralization using MC3T3 cells and testing the hypothesis that mineralization is inhibited via alteration of the Enpp1–TNAP–Ank axis. Inhibition of mineralization was not due to disruption of collagen deposition or calcium chelation by the negatively charged pVTK. The timing of peptide administration was important in inhibiting mineralization – pVTK had a greater effect at later stages of osteogenic differentiation (days 7–12 of culture corresponding to matrix maturation and mineralization), and could prevent progression of mineralization once it had started. pVTK treatment resulted in a significant decrease in ectonucleotide pyrophosphatase/phosphodiesterase 1 (Enpp1) enzyme activity and gene expression. The expression of ankylosis protein (Ank), osteopontin (OPN) and Pit-1 genes was also significantly reduced with peptide treatment, while tissue non-specific alkaline phosphatase (TNAP), bone sialoprotein (BSP), and Runx2 gene expression was significantly higher. The ability of pVTK to inhibit mineralization can potentially be translated into therapeutics against pathological calcification seen in cardiovascular disease, osteoarthritis or craniosynostosis, or be used to prevent failure of biomaterials due to calcification, such as bioprosthetic heart valves.

English

This study aimed to determine the Pit-1|Hinf-1 gene polymorphism in Bali cattle ( Bos javanicus ) as Indonesian native cattle and besides Madura, Pesisir, Aceh, and Katingan cattle breeds as a comparison. DNA samples were extracted from 488 blood samples consisting of Bali (245 heads), Madura (68 heads), Aceh (25 heads), Pesisir (100 heads) and Katingan (50 heads) cattle. The diversity of the Pit-1|Hinf-1 gene wasere analyzed using PCR-RFLP. Whereas the nucleotide base mutations were identified by sequencing. Genotyping data were analyzed by calculating the allele frequency, observed heterozygosity (Ho) and expected heterozygosity (He) values as well as Hardry-Weinberg equilibrium test using POPGENE 1.31 program. Whereas, Tthe sequence data were analyzed by using MEGA6 program. The Pit-1|Hinf-1 gene fragment analysis showed that Bali, Madura, Pesisir, Aceh, and Katingan cattle had high BB genotype, resulting in B allele frequency of 0.982, 0.963, 0.925, 0.960, and 0.960, respectively. Ho and He values were 0.074-0.130 and 0.036-0.139, respectively. Hardy-Weinberg equilibrium test did not significant for all breed populations, except for Aceh cattle population (P<0.05). Mutation from guanine (G) to adenine (A) was found in Pit-1 gene fragment. Therefore, Pit-1|Hinf-1 gene fragment had low genetic diversity in Bali cattle and other breeds population. Key words: Bali cattle, Pit-1|Hinf-1 gene, polymorphism

Allele substitution effects of IGF1, GH and PIT1 markers on estimated breeding values for weight and reproduction traits in Canchim beef cattle

The aim of this study was to evaluate the allele substitution effects of IGF1, GH and PIT1 markers on the estimated breeding values (EBVs) for birth (BW) and weaning weights (WW), weight at 12 (W12) and 18 (W18) months of age, weight (WFC) and age (AFC) at first calving, and scrotal circumference measured at 12 (SC12) and 18 (SC18) months of age in Canchim beef cattle. Maternal effects were considered for birth (BWmaternal) and weaning (WWmaternal) weight. Regression analyses were carried out considering the EBVs, obtained from multi-trait analyses, and the deviations for each allele studied (four alleles for the IGF1 markers and two for GH and PIT1 markers, respectively). According to the results obtained for IGF1, the substitution effect of the “225” allele was significant (P≤0.05) and presented higher EBVs for BW, BWmaternal, WW, W12, W18, and SC18; and lower EBVs for WWmaternal. The allele substitution effect observed for GH gene was significant (P≤0.05) on EBVs for AFC and WFC. The “Valine” allele was responsible for lower EBVs for AFC and higher EBVs for WFC. For the PIT1 gene, the substitution effect of the “Hinf−” allele was significant (P≤0.05) on the EBVs for WW, WWmaternal, AFC, and WFC, respectively. Important allele substitution effects were found for weight and reproduction traits in Canchim cattle. In general, markers on the IGF1 gene are related to higher EBVs for weight, while markers on the GH and PIT1 showed greater influence on the EBVs for age and weight at first calving. Adding molecular markers information on the selection process could result in increased genetic gains in Canchim cattle. Future studies using high-density genotyping platforms may contribute to our understanding of the effects of these genes on traits of economic importance in the Canchim breed.

Association of PIT-1 gene polymorphism with milk production traits in Sahiwal cattle

In the present study, identification of PIT-1 gene polymorphism and its association with milk production traits was undertaken in 77 Sahiwal cattle maintained at NDRI, Karnal using PCR-RFLP. Amplification of DNA sample revealed 600 bp product and restriction digestion with HinfI showed three types of genotypes, namely, AA (600bp), AB (600, 343 and 257 bp) and BB (343 and 257 bp) genotypes. The frequency of BB genotypes was highest (64.98%) in all screened samples, followed by AB genotype (31.16%) and AA genotype (3.89%). The allelic frequency of PIT-1 A and B alleles were 19.48% and 80.51%, respectively. Association studies of PIT-1 gene with milk production traits showed that age at first calving, gestation period and lactation period had non-significant variation among all the 3 genotypes up to fourth lactation. However, a significant difference was found among the 3 genotypes for total milk yield and milk yield at 300 days with AA genotype showing higher value than AB and BB genotypes in the first lactation. From present investigation, the HinfI /PCR-RFLP revealed polymorphic pattern of PIT-1 gene in Sahiwal cattle and association studies showed significant effect of A allele on milk production.

Polymorphism of Prophet of Pit-1 gene and its relationship with litter size of Small Tail Han sheep.

1Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, People’s Republic of China 2College of Life Sciences, Anhui Agricultural University, Hefei 230036, People’s Republic of China 3College of Life Science, Liaoning Normal University, Dalian 116029, People’s Republic of China 4College of Animal Science and Technology, Agricultural University of Hebei, Baoding 071000, People’s Republic of China 5State Key Laboratory of Agricultural Biotechnology (China Agricultural University), Beijing 100193, People’s Republic of China