PRLR Gene Research Articles

Maintaining Breed Integrity: Successful Introgression of the SLICK1 Allele into the Holstein Breed.

This study evaluated the effectiveness of genetic introgression of the SLICK1 allele derived from Senepol cattle into the Holstein breed to enhance thermotolerance. The SLICK1 allele, located in PRLR gene, confers a short and sleek coat that is inherited as a simple dominant phenotype. Approximately 40 years ago, the University of Florida initiated efforts to introgress this allele into the Holstein population. Here we tracked the introgression of the SLICK1 allele using a medium-density genotyping array and a reference population of both breeds (50 Holstein, 46 Senepol). Among the 31 SLICK1+ Holsteins, there was 15.25% ± 11.11% (mean ± SD) Senepol ancestry on BTA20. Holsteins at the University of Florida descended from slick matings that did not inherit the SLICK1 allele (n=9) exhibited no Senepol ancestry. A secondary introgression of Senepol genetics in SLICK1+ animals was found on BTA4, spanning 54 markers and 15 genes, with 26.67% Senepol ancestry. This region, previously linked to heat stress adaptation, suggests that the introgression extends beyond the SLICK1 allele to incorporate additional beneficial genetics for thermal stress adaptation. These findings indicate that deliberate introgression of the SLICK1 allele enhances specific traits and potentially introduces other adaptive genetic variations. The study demonstrates the successful use of genetic interventions to improve livestock resilience against environmental challenges without significantly disrupting the recipient breed's genetic structure. The introgression of the SLICK1 allele serves as a model for breeding programs aimed at optimizing animal welfare and productivity in the face of global climate change while maintaining breed integrity.

PSV-19 Uncovering breed-origin allele effects and their role in thermotolerance through genome-wide association study in Brangus cattle

Abstract In subtropical and tropical environments, beef cattle production faces significant challenges due to heat stress, affecting animal welfare and productivity. To mitigate these effects, producers have turned to crossbreeding and the development of composite breeds like Brangus, which combine the desirable carcass and meat quality traits of European taurine cattle with the thermotolerance of Indicine breeds. However, the genetic underpinnings of thermotolerance in these composite populations are not well understood, owing to the complex nature of measuring related traits and the genetic intricacies arising from crosses of distinct populations. This study aims to identify key quantitative trait loci (QTL) related to thermotolerance, employing an approach that integrates genetic markers alongside the marker’s breed of origin of alleles (BOA) in Brangus heifers. Data were collected on histological sweat gland area (SWA), hair length (HL), and thermal stress slope as a measure of the increase in body temperature from a low to a high temperature-humidity index (TSS), from 2,233 Brangus heifers genotyped with the 250K functional chip. BOA was determined using LAMP-LD software, using Angus and Brahman cattle from the University of Florida’s multibreed Angus-Brahman project as a reference population. We conducted a genome-wide association study (GWAS) incorporating BOA-specific effects, revealing several genes with BOA-specific effects. Notably, the CGGBP1 gene exhibited a significant effect on TSS only when alleles originated from Brahman. Conversely, the PLK1 gene was significant for TSS when alleles originated from Angus. Both CGGBP1 and PLK1 interact with heat shock proteins during heat stress, these findings reveal different genetic mechanisms for controlling internal body temperature. For HL, a significant Brahman-specific effect was found near the PRLR gene, which is known as the gene controlling the SLICK hair phenotype in Criollo cattle. For SWA, an Angus-specific significant association was found with the PDE4D gene. PDE4D interacts with the cAMP pathway which regulates sweat gland growth and sweat production. These findings underline the genetic complexity of thermotolerance in composite cattle and the crucial role of BOA in identifying key QTL. The study offers novel insights into the genetic mechanisms of thermotolerance, providing a foundation for breeding strategies aimed at improving heat resilience in cattle in subtropical and tropical environments.

Effect of prolactin concentration during the dry period on the subsequent milk production of dairy cows

Shortening the dry period has a negative impact on milk production of the following lactation. One possible explanation is that a period of low prolactin (PRL) concentration is necessary to restore mammary gland milk production capacity. Therefore, the objective of this study was to determine the effect of lowering blood PRL level on subsequent lactation milk production. In this experiment, quinagolide was used to inhibit PRL secretion during the dry period. Thirty Holstein cows were randomly assigned one of 3 dry period managements: a conventional (60 d) dry period (CD) and 2 short (35 d) dry period treatments (SD). Short dry period cows received either water (SDwater) or quinagolide (2 mg, SDquin) injections twice daily from dry-off until 14 d before calving. Cows were followed during the first 20 wks of the subsequent lactation. When CD cows were dry but SD cows were lactating, concentration of PRL was lower in the CD cows than in the SD cows. During the injection period, PRL of SDquin cows was lower than that of the other treatments and was greater in the blood of SDwater than in that of CD cows. After the injection period until calving, no difference in PRL concentration was observed between treatments. After calving, PRL concentration of the SDquin cows was greater than those of CD and SDwater cows. During the first 20 wks of lactation, energy corrected milk (ECM) was lower in SDwater cows than in CD and SDquin cows. The ECM of the latter groups were not different. During the same period, mammary expression of genes related to milk synthesis, pro-apoptotic genes, as well as the expression of the short and long isoforms of the PRLR genes were not affected; however, the expression of SOCS3 gene tended to be lower for the SDquin than the SDwater cows. Lowering the PRL level during short dry period restored milk production to the level normally observed after a conventional dry period, which suggests that higher PRL levels during short dry period are the cause of the lower milk production after a short dry period. Ultimately, strategies to lower blood PRL level may help the adoption of short dry period.

Significance of Duodenal Prolactin Receptor Modulation by Calcium and Vitamin D in Sulpiride-Induced Hyperprolactinemia.

Background and Objectives: Hyperprolactinemia, as a potential side-effect of some antipsychotic medications, is associated with decreased bone density and an increased risk of fractures. This study investigates whether calcium and vitamin D supplementation affects prolactin receptor (Prlr) gene expression in the duodenum, vertebrae, and kidneys of female rats with sulpiride-induced hyperprolactinemia. Materials and Methods: Twenty-one-week-old female Wistar rats were assigned to three groups: Group S consisted of ten rats who received sulpiride injections (10 mg/kg) twice daily for 6 weeks; Group D (10 rats) received daily supplementation of 50 mg calcium and 500 IU vitamin D along with sulpiride for the last 3 weeks; and Group C consisting of seven age-matched nulliparous rats serving as a control group. Real-time PCR was used to assess Prlr gene expression in the duodenum, vertebrae, and kidneys. Results: In Group S, Prlr gene expression was notably decreased in the duodenum (p < 0.01) but elevated in the vertebrae and kidneys compared to Group C. Conversely, Group D exhibited significantly increased Prlr expression in the duodenum (p < 0.01) alongside elevated expression in the vertebrae and kidneys. Conclusions: In sulpiride-induced hyperprolactinemia, decreased Prlr gene expression in the duodenum may lead to reduced intestinal calcium absorption. Consequently, prolactin may draw calcium from the skeletal system to maintain calcium balance, facilitated by increased Prlr gene expression in the vertebrae. However, vitamin D supplementation in sulpiride-induced hyperprolactinemia notably enhances Prlr gene expression in the duodenum, potentially ameliorating intestinal calcium absorption and mitigating adverse effects on bone health.

The prolactin receptor: A cross-species comparison of gene structure, transcriptional regulation, tissue-specificity, and genetic variation.

The conserved and multifaceted functions of prolactin (PRL) are coordinated through varied distribution and expression of its cell-surface receptor (PRLR) across a range of tissues and physiological states. The resultant heterogeneous expression of PRLR mRNA and protein across different organs and cell types supports a wide range of PRL-regulated processes including reproduction, lactation, development, and homeostasis. Genetic variation within the PRLR gene also accounts for several phenotypes impacting agricultural production and human pathology. The goal of this review is to highlight the many elements that control differential expression of the PRLR across tissues, and the various phenotypes that exist across species due to variation in the PRLR gene.

Preliminary monosodium glutamate-induced changes in mammary gland receptors and gene expression, water channel, oxidative stress, and some lactogenic biomarkers in lactating rats

BackgroundChanges induced by monosodium glutamate (MSG) can negatively impact milk production and secretion, among other adverse effects. This study aimed to investigate the effects of MSG consumption on receptor gene expression and quantification of hormones and receptors, as well as oxidative stress biomarkers and other lactogenic parameters in lactating animals. Twenty-four female Wistar rats, nine weeks of age, were randomly assigned to four groups, each containing six rats, at parturition. The rats in groups II, III, and IV were given varying doses of monosodium glutamate (MSG); while, group I was given distilled water and served as the control. The experimental period lasted two (2) weeks.ResultsThe groups administered with MSG showed a significant decrease in mammary PRLR gene expression (p < 0.05), as well as a marked reduction (p < 0.05) in mammary PRLR, OXT receptor, AQP-3, brain antioxidant enzymes (SOD, GPx, and CAT), and pituitary SOD compared to the control group (p < 0.05). Furthermore, there was a significant increase (p < 0.05) in reactive oxygen species levels in the serum and mammary gland homogenates, erythrocyte osmotic fragility, and elevated (p < 0.05) brain and pituitary MDA levels in the MSG-administered groups compared to the control group. Daily milk yields were significantly decreased (p < 0.05) in the MSG-administered groups between days 10 and 14 of lactation.ConclusionThe findings of this study suggest that prolonged consumption of MSG could interfere with lactation-associated functions via increased ROS production, reduced antioxidants, decreased AQP-3, mammary prolactin and oxytocin receptors, and prolactin receptor mRNA in lactating Wistar rats.Graphical

Identification of genomic regions associated with resistance to gastrointestinal parasites in an indigenous sheep by single- and multiple-locus methods.

We investigated the association between 157 SNPs located in 75 candidate genes involved in the immune system and proxy traits for resistance to gastrointestinal nematodes in sheep. A total of 211 lambs from eight flocks were sampled. Nematode eggs per gram were counted and classified as: (i) Strongyles, (ii) Nematodirus spp., (iii) Trichuris spp. and (iv) Marshallagia marshalli. Single- and multiple-locus models were used to test the marker-trait associations. Seven significant SNPs were identified on chromosomes OAR6, 15, 16, and 19. These findings provide insights for breeding nemarode-resistant traits in low-input production systems. General linear model, fixed and random model circulating probability unification, and Bayesian-information and linkage-disequilibrium iteratively nested keyway analyses identified a significant association between the eggs per gram of Strongyles nematodes and a specific variant of the PRLR gene.

Milk-y Way: the impact of single-nucleotide polymorphisms on milk production traits in Kilis dairy goats.

This study aimed to investigate the impact of single-nucleotide polymorphisms (SNPs) on milk production traits in Kilis dairy goats by analyzing the genotypes of POU1F1, PRLR, -Lg, GH1, and GH2 genes and their association with lactation milk yield (LMY), lactation length (LL) and average daily milk yield (ADMY). Blood samples were collected from 227 goats, and genotyping was performed using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The results revealed that the frequencies of the genotypes varied among the genes. The polymorphisms were found to be significantly linked with milk production traits. These findings suggest that SNPs of POU1F1/AluI, PRLR/RsaI and -Lg/SacII are significantly associated with LMY and that the POU1F1-TC genotype, PRLR-TT genotype and -Lg-AB genotype are associated with higher LMY and ADMY. Additionally, the POU1F1-TC genotype was found to have a longer LL. However, no significant association was found between the GH1 and GH2 genotypes and LMY, LL and ADMY. Overall, this study provides valuable insights into the genetic factors influencing milk production traits in Kilis dairy goats, which can be utilized for the selection of high-yielding animals in breeding programs.

The prolactin receptor gene (PRLR) is linked and associated with the risk of polycystic ovarian syndrome

The prolactin receptor gene (PRLR) may contribute to polycystic ovarian syndrome (PCOS) since it plays important roles in physiological ovarian functions. PRLR-knockout mice have irregular cycles and subfertility and variants in or around the PRLR gene were associated in humans with female testosterone levels and recurrent miscarriage. We tested 40 variants in the PRLR gene in 212 Italian families phenotyped by type 2 diabetes (T2D) and PCOS and found two intronic PRLR-variants (rs13436213 and rs1604428) significantly linked to and/or associated with the risk of PCOS. This is the first study to report PRLR as a novel risk gene in PCOS. Functional studies are needed to confirm these results.

FRI303 PRL-R Variants Are Not Only Associated With Prolactinomas But Also With Dopamine Agonist Resistance

Abstract Disclosure: A.R. Moreira: None. E.B. Trarbach: None. A. Glezer: None. G.A. Maciel: None. C.B. Bueno: None. A.S. Monteiro: None. M. Padula: None. I.P. Grande: None. M.D. Bronstein: None. Introduction: Knockout prolactin receptor gene (PRL-R) mice are animal models for prolactinomas and PRL acts via autocrine/paracrine inhibiting lactotroph proliferation. Recent studies identified variants in PRL-R gene in patients with prolactinoma and two of them Glu400(376)Gln/p.Asn516(492)Ile, were more common in patients compared to controls. Functional analysis of the p.Asn516(492)Ile variant showed an increase in Akt expression and cell proliferation induced by PRL. However, the association between PRL-R variants and response to cabergoline (CAB) treatment has not been investigated. Objective: This study aims to analyze the PRL-R gene in an extensive cohort of patients with sporadic prolactinomas and evaluate its association with clinical, laboratorial and tumor imaging characteristics, as well as CAB treatment resistance. Methods: DNA was extracted from peripheral blood samples of 178 patients with prolactinoma and 146 were classified according to cabergoline response. The PRL-R gene was analyzed by PCR techniques and automated Sanger-type sequencing. Four in silico tools were used for analysis. Results: We found six PRL-R variants: p.Ile100(76)Val (n=18), p.Ile170(146)Leu (n=6), p.Glu400(376)Gln/p.Asn516(492)Ile (n=3), p.Glu470Asp (n =1) and p.Ala591Pro (n=1), the last two described for the first time in prolactinomas´ patients. The variants p.Glu400(376)Gln/p.Asn516(492)Ile and p.Ala591Pro were more frequent amongst patients compared to genomic databases and the p.Asn516(492)Ile showed pathogenic potential using in silico analysis. PRL-R variants were associated with male sex (P=0.015), higher serum PRL levels (P=0.007), larger tumors (P=0.001), higher frequency of macroadenomas (P=0.031) and cabergoline resistance (P&amp;lt;0.001). Considering together only the PRL-R variants classified as pathogenic/probably pathogenic (p.Ile170(146)Leu, p.Glu400(376)Gln/Asn516(492) and p.Ala591Pro) versus the benign variants (p.Ile100(76)Val, p.Glu470Asp and wild type) we also observed a higher levels of serum PRL (P=0.008), largest tumor diameter (P=0.028) and higher frequency of CAB resistance (P=0.048).Conclusion: Our study showed a higher frequency of the PRL-R variants p.Glu400(376)Gln/p.Asn516(492)Ile, and p.Ala591Pro in patients with prolactinomas when compared to Brazilian and international databases, suggesting that these variants could play a role in the prolactinoma tumorigenesis. Patients with PRL-R variants presented male predominance, higher serum PRL levels, larger tumor size at diagnosis and higher frequency of CAB resistance. More studies evaluating germinative and somatic PRL-R variants are needed to clarify the PRL/PRL-R system role in the tumorigenesis and progression of prolactinomas and their interesting association with CAB resistance. Presentation: Friday, June 16, 2023

A precise molecular subtyping of ulcerative colitis reveals the immune heterogeneity and predicts clinical drug responses

Background and AimsWe sought to identify novel molecular subtypes of ulcerative colitis (UC) based on large-scale cohorts and establish a clinically applicable subtyping system for the precision treatment of the disease.MethodsEight microarray profiles containing colon samples from 357 patients were utilized. Expression heterogeneity was screened out and stable subtypes were identified among UC patients. Immune infiltration pattern and biological agent response were compared among subtypes to assess the value in guiding treatment. The relationship between PRLR and TNFSF13B genes with the highest predictive value was further validated by functional experiments.ResultsThree stable molecular subtypes were successfully identified. Immune cell infiltration analysis defined three subtypes as innate immune activated UC (IIA), whole immune activated UC (WIA), and immune homeostasis like UC (IHL). Notably, the response rate towards biological agents (infliximab/vedolizumab) in WIA patients was the lowest (less than 10%), while the response rate in IHL patients was the highest, ranging from 42 to 60%. Among the featured genes of subtypes, the ratio of PRLR to TNFSF13B could effectively screen for IHL UC subtype suitable for biological agent therapies (Area under curve: 0.961–0.986). Furthermore, we demonstrated that PRLR expressed in epithelial cells could inhibit the expression of TNFSF13B in monocyte-derived macrophages through the CXCL1-NF-κB pathway.ConclusionsWe identified three stable UC subtypes with a heterogeneous immune pattern and different response rates towards biological agents for the first time. We also established a precise molecular subtyping system and classifier to predict clinical drug response and provide individualized treatment strategies for UC patients.

Analysis of polymorphic variants of MSTN, CAST, PRLR genes associations with economically useful qualities of vyatka breed horses

The relationship of polymorphic variants of MSTN, CAST, PRLR genes with working qualities and body types of Vyatka horses was studied. The purpose of the research is to assess the genetic and breeding potential of the Vyatka breed horses, as well as to study the relationship between the MSTN, CAST, PRLR genotypes and the economically useful qualities of horses. The method of DNA extraction from Vyatka horse hair follicles using ExtraGene DNA Prep. was applied. When scanning the mutations in the loci MSTN (n=43), CAST (n=41) and PRLR (n=41), DNA amplification the method of allele-specific PCR was performed. The frequency of alleles and genotypes was calculated using MS Excel 10. As a result of research, the dependence of Vyatka horses working qualities on the frequency of occurrence of myostatin MSTN alleles (g.66493737 T&gt;C) was noted. The horses with a higher occurrence of the MSTN/C allele have more productive movements than the individuals with the T/T genotype typical for aborigines. Vyatka horses with the T/T genotype are more versatile, and also show better results in sledding, while the horses with the T/C genotype are better under saddle. The horses with the T/T genotype have the highest bony index but the lowest massiveness index. The MSTN T/T (0.581) homozygous genotype for the "wild" allele predominates in Vyatka horses. A relation ship between body types and the calpastatin gene (CAST) has been revealed for the first time in horse breeding. The horses with the G/A genotype turned out to be the most massive and bony, the horses with the A/A genotype were lighter, the highest frequency of occurrence of the CAST G/A genotype (0.463) was noted, the CAST G/G genotype is rare in the breed (0.171). A relationship between the frequency of occurrence of prolactin receptor genes (PRLR) and body types of horses was not found. The frequency of occurrence of PRLR C/C (0.366) and PRLR G/C (0.390) genotypes is approximately identical, the PRLR G/G genotype is less common (0.244). The study of genes associated with economically useful qualities in all breeding stallions will enable to conduct more efficient breeding, using the desired genotypes, which is important for small breeds.

Results of Targeted Sequencing of PRL, PRLR, and PRLHR Genes in Young Women with Nonneoplastic Hyperprolactinemia

Results of Targeted Sequencing of PRL, PRLR, and PRLHR Genes in Young Women with Nonneoplastic Hyperprolactinemia

Genetic diversity of porcine PRLR gene and its relationship to litter size in Large White pigs

Improving the litter performance of sows is one of the main challenges in the current pig industry. In this study, single nucleotide polymorphisms (SNPs) of the PRLR gene were performed, in order to test whether they are associated with the reproductive performance of Large White pigs. In total, we discovered nine SNP loci (g. C260G, g. C362T, g. C527G, g. A540G, g. A584G, g. A673T, g. A745G, g. C765T and g. A934G) in exon 10 of PRLR. The result showed that genotypes CC and CT at the g. C362T locus and genotype AG at g. A584G could significantly increase the litter size of different strains of Large White pigs (p &lt; 0.05). In addition, the genotype CC at the g. C765T locus and the genotype AA at g. A934G could also increase the litter size (TNB could be increased by 1.5 piglets per year; while NBA could be increased by 0.98 piglets per year, p &lt; 0.01). Furthermore, the haplotype combinations of H2H7 and H4H4 were the dominant combinations and contributed to larger litter sizes in the Large White pigs. In conclusion, there were dominant genotypes in the related SNP loci in the PRLR gene that were beneficial to improving the litter traits of sows. These findings will provide a reference for screening the molecular markers of a high reproductive performance in sows, and are helpful for genetic breeding and the reproductive improvement of pigs. Key words: sow, prolactin, polymorphism, reproduction, association analysis.

The Role of PRLR Gene Polymorphisms in Milk Production in European Wild Rabbit (Oryctolagus cuniculus).

One of the problematic points of rabbit breeding is that the nutritional requirements of the kits are not fully satisfied by the does' milk production from the third week of lactation onwards. The prolactin receptor gene has a significant effect on reproductive processes, and its polymorphisms have been associated with milk production in several species (cattle, goats, sheep, and buffalo). The European wild rabbit (Oryctolagus cuniculus), has a more diverse genetic background compared to domesticated lines. In the course of our study, sequencing of the 1210 bp long segment of the PRLR gene promoter region was accomplished. We detected four point mutations (SNP1-407G > A, SNP2-496G > C, SNP3-926T> and SNP4-973A > C) and one microsatellite at position 574. In our population, the four SNPs were segregated into four genotypes: AACCCCCC, GGGGTTAA, AAGGTTAC, and GGGGTCAC. Our results show that the genotype in the homozygous form is associated with higher milk production (1564.7 ± 444.7 g) compared to the other three genotypes (AACCCCCC 1399.1 ± 326.8 g; GTGACCTT 1403.8 ± 517.1 g; GGGGTCAC 1220.0 ± 666.2 g), and the short microsatellite repeat (167 bp) also coincides with significantly higher milk production (1623.8 ± 525.1 g). These results make the marker-assisted selection possible also for domesticated lines.

Molecular characterisation and expression profile of the PRLR gene during goose ovarian follicle development

ABSTRACT 1. Although PRL-PRLR signalling plays important roles in regulating avian reproduction, there is a paucity of information regarding the functional significance of PRLR in goose ovarian follicle development. 2. The full-length 2,496 bp coding sequence of PRLR was obtained from Sichuan White goose (Anser cygnoides) for the first time and was seen to encode a polypeptide containing 831 amino acids. Goose PRLR shares similar sequence characteristics and conserved functional domains to other avian species and was phylogenetically clustered into the avian clade. 3. The qPCR results suggested that the mRNA levels of PRLR significantly increased in primary follicles during weeks 3 to 4 of age and were higher in secondary- than in primordial follicles at week 5 post-hatching, which suggested that the PRLR-mediated signalling could be involved in regulation of early folliculogenesis. 4. The PRLR mRNA was expressed at the highest levels in the prehierarchical 8–10 mm granulosa layers throughout goose ovarian follicle development, indicating a role for PRLR in the process of follicle selection. 5. PRLR mRNA was differentially expressed in the three cohorts of in vitro cultured granulosa cells harvested from different sized goose ovarian follicles, which suggested that PRLR was involved in regulating granulosa cell functions depending on the stage of follicle development. These data provide novel insights into the role of PRLR during goose ovarian follicle development, although the underlying mechanisms await further investigations.

Expression Variance of the PRLR Gene and its Association with Milk Productive Traits in Pure Holstein-Friesian and Montbeliarde×Holstein Crossbred Cattle Raised under Egyptian Condition

The objective of this study was to investigate the association between the expression level of the PRLR gene and milk production traits at different parities in pure Holstein-Friesian and Montbeliarde×Holstein crossbred cattle reared under Egyptian conditions, as well as its potential use in future breeding programs. For this purpose, eighteen healthy cows in mid lactation stage belonging to the first, second, and third parities were selected from each breed for sampling, RNA extraction, and quantitative real-time PCR. The results revealed that, the mean relative expression of prolactin receptor gene and its fold change level were significantly higher in pure Holstein-Friesian than Montbeliarde×Holstein crossbred at the level of the first three parities. In addition, there is a significant and strong positive correlation between PRLR1 and PRLR3 in pure Holstein-Friesian cattle. Significant negative correlations between PRLR2 and PRLR3, DMY3 and TMY3, were observed in Montbeliarde×Holstein crossbred cows. In conclusion, this information about the PRLR gene could be a useful indicator for the milk production trait and could be used in crossing programs that depend on genetic selection.

Changes of proapoptotic and antiapoptotic genes affect sensitivity to apoptotic stimuli in impaired contractility due to long term bladder outlet obstruction.

The normal biological process that necessitates cell removal greatly depends on apoptosis. Long term bladder outlet obstruction (BOO) causes damaged smooth muscle cells to undergo apoptosis. However, smooth muscle cell apoptosis that BOO causes is not well known in impaired bladder contractility. Therefore, we designed this study to investigate whether long-term BOO could induce apoptosis activities and to obtain an expression profile of apoptosis related genes. We used 10 Sprague-Dawley six-week-old female rats. We separated them equally into two groups: a sham intervention group (group 1) and an eight-week BOO group (group 2). We conducted cystometric evaluation eight weeks following BOO onset, with processing of bladder tissue for PCR array. Every array comprised 84 genes, which were established to contribute to an apoptosis response, cell differentiation and metabolism, and 12 sequences were established for the regulation of loading and the quality of cDNA. We performed real-time PCR. Changes in gene expression presented as a fold increase/decrease. Alterations of more than two-fold constituted the cut-off determining expression. Group 2 had a greater bladder weight and Impaired bladder contractility. Immunofluorescent staining with CAS3, TUNEL showed increased in the BOO group. In comparison to group 1, group 2 exhibited an at least two-fold upregulation in five genes, the Bcl-2 (15.1), Birc5 (5.8), Cd40lg (7.5), Il10 (16.2), and Naip2 (13.2). They also demonstrated at least a two-fold downregulation in the PRLR (-18.1) gene. Genes Bcl2ald, Circ5, Cd40lg, Il10, Naip2, and PRLR were among the genes with activity against apoptosis. TNF, STAT3 and TP53 mediated the effect that genes had on one another. This study demonstrated that the relative ratios of pro- and antiapoptotic genes determine bladder cell sensitivity cells to apoptotic stimuli in impaired contractility caused by long term BOO. Although we cannot confirm whether this finding is the result of the decompensated phase of the bladder or the process, the gene expression profiles could explain molecular mechanisms of apoptosis in impaired bladder contractility caused by long-term BOO with further studies.

PSIII-8 Evaluation of Reproductive Efficiency of Holstein Nulliparous Heifers Carrying the Slick1 Mutation of the Prolactin Receptor (Prlr) Gene

Abstract Our objective was to evaluate the reproductive performance of Holstein nulliparous heifers carrying the SLICK1 mutation and half-sisters not carrying the mutation. Heifers were born between November and March of 2020 to two Holstein sires heterozygous for the dominant SLICK1 allele and raised in four dairy farms in the San Joaquin and the Tulare basins of the California Central Valley. Heifers were bred from January to September of 2021 according to farm protocols. Data were obtained via farm records and analyzed with the MIXED and LOGISTIC procedures of SAS; proportions were obtained via the FREQ procedure. Models included the fixed effects of genotype, sire, and farm; animal was considered a random effect. Response variables included overall pregnancy rate, age at first breeding, services per conception, pregnancy rate to first insemination, calving rate and pregnancy length. As shown in Table 1, pregnancy and calving rates were similar between slick and non-slick heifers. Overall, services per conception were similar between slick and non-slick heifers (1.8±0.2 and 2±0.2 services for slick and non-slick heifers, respectively; P = 0.3); however, pregnancy to first service tended to be greater in slick heifers (P = 0.07). Age at first breeding was not affected by genotype or sire, but there was an effect of farm due to one dairy starting the breeding program at an older heifer age. Pregnancy length tended to be longer for slick heifers (P = 0.08). This is the first study to evaluate the reproductive performance of slick Holstein females, although it is limited in sample size. Results indicate that the presence of the SLICK1 allele could provide an advantage to reproductive efficiency of Holstein heifers by improving the rate of pregnancy to first service. The authors thank the participating dairy farms. Support: Holstein Association USA Research Grant Program.

Results of targeted sequencing of the &lt;i&gt;PRL, PRLR, PRLHR&lt;/i&gt; genes in young women with non-tumor hyperprolactinemia

Aim. To study the spectrum of variants in the PRL, PRLR, PRLHR genes in women of reproductive age with non-tumor hyperprolactinemia. Material and methods. In women with non-tumor hyperprolactinemia (n = 15), targeted high-throughput sequencing of the PRL, PRLR, and PRLHR genes was performed. The target panel of genes included coding regions and adjacent splicing sites. Results. When analyzing the PRL, PRLR, PRLHR genes, a number of rare and common variants were identified. The common variant rs1205955 was found in the PRL gene (MAF А = 0.279). For the PRLR gene, a rare variant rs185353023 was identified in the 3’UTR (MAF А/С = 0.003) and 12 common variants. For the PRLHR gene, 10 common variants have been identified. The maximum number of variants was localized in the 3’UTR region and introns. Conclusions. For the first time in Russia, targeted high-throughput sequencing of the PRL, PRLR, PRLHR genes was performed, the results of which did not reveal obvious pathological variants in the studied genes in women with high prolactin content of non-tumor origin. The discovered polymorphism in these genes makes it possible to further study its association with impaired function of the prolactin link of hormonal regulation.