Scholarly Works, School of Animal Sciences

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  • A meta-analysis of the relationship between milk protein production and absorbed amino acids and digested energy in dairy cattle
    Mark Hanigan; Vinícius Carneiro de Souza; Roger Martineuau; Hélène Lapierre; Xin Feng; Veridiana Daley (Journal of Dairy Science)
    Supplemental materials of the paper "A meta-analysis of the relationship between milk protein production and absorbed amino acids and digested energy in dairy cattle".
  • Fetal Programming and Its Effects on Meat Quality of Nellore Bulls
    Christofaro Fernandes, Arícia; Beline, Mariane; Polizel, Guilherme Henrique Gebim; Cavalcante Cracco, Roberta; Ferreira Dias, Evandro Fernando; Furlan, Édison; da Luz e Silva, Saulo; de Almeida Santana, Miguel Henrique (MDPI, 2023-11-24)
    This work aimed to evaluate the effects of prenatal nutritional stimulation at different pregnancy stages on carcass traits and meat quality in bovine progeny. For this purpose, 63 Nellore bulls, born from cows submitted to three nutritional plans, were used: not programmed (NP), which did not receive protein supplementation; partially programmed (PP), which had protein-energy supplementation (0.3% of mean body weight of each batch) only in the final third of pregnancy; and full programming (FP), which received supplementation (0.3% of mean body weight of each batch) throughout pregnancy. The averages of parameters were submitted to the ANOVA, and the supplementation periods, which were different when p value < 0.05, were compared. Carcass weights and rib eye area (REA) did not differ between treatments (p > 0.05), but subcutaneous fat thickness (SFT) showed a tendency (p = 0.08) between groups. For lipids and marbling, no differences were found (p > 0.05). In the analyses of maturation time and shelf life, no difference was observed between treatments. However, there was a tendency between treatments at 14 days of maturation time for cooking loss (CL) (p = 0.08). Treatments did not affect shear force in the progenies (p > 0.05). Fetal programming had no effect on the meat quality of Nellore bulls.
  • Farm-Scale Effectiveness of Feed Additives Supplied through a Mineral Mix for Beef Cattle Grazing Tropical Pastures
    Goulart, Ricardo Cazerta Duarte; Costa, Diogo Fleury Azevedo; Silva, Tiago Alves Corrêa Carvalho da; Congio, Guilhermo Francklin de Souza; Marques, Rodrigo da Silva; Corsi, Moacyr (MDPI, 2023-12-13)
    The effectiveness of feed additives delivered through free-choice mineral mixtures (MMs) to grazing cattle remains unclear. Two farm-scale and one in vitro experiment (Exp.) were conducted to investigate the effects of salinomycin and virginiamycin, delivered through an MM, on growing bulls grazing tropical pastures. In Exp. 1, 316 zebu (Bos indicus) Nellore bulls (225 ± 26.7 kg liveweight (LW)) were randomly allocated to four treatments: (1) MM no additives (CON), (2) MM with salinomycin at 1950 mg/kg (SLI), (3) MM with salinomycin at 780 mg/kg (SHI), and (4) MM with virginiamycin at 1950 mg/kg (VGN). Over 123 days, these bulls grazed tropical grasses on pastures of guinea grass, palisade grass, or Bermuda grass. No significant treatment effects were observed for oocyst eggs or ruminal parameters. Bulls fed VGN had higher average daily gain (ADG) compared to CON (p = 0.02) and SLI (p = 0.03) but similar compared to SHI (p = 0.07). In Exp. 2, 308 zebu cross bulls (237 ± 23.0 kg LW) grazed Bermuda grass paddocks and were allocated into two treatments: (1) MM with no additives (CON) and (2) MM containing virginiamycin at 2522 mg/kg (VGN). Cattle fed VGN had a significantly higher ADG (p = 0.007). Exp. 3 tested salinomycin’s effectiveness in vitro at different exposure times to MM, revealing no impact of exposure time on short-chain fatty acid production. In conclusion, virginiamycin delivered through free-choice MM can increase grazing beef bulls’ ADG by 12% compared with CON, with no clear link to rumen fermentation or coccidiostat effects.
  • Influences of Supplementing Selective Members of the Interleukin-6 Cytokine Family on Bovine Oocyte Competency
    McKinley, Endya; Speckhart, Savannah L.; Keane, Jessica A.; Oliver, Mary A.; Rhoads, Michelle L.; Edwards, J. Lannett; Biase, Fernando H.; Ealy, Alan D. (MDPI, 2023-12-21)
    This work explored whether supplementing selective members of the interleukin-6 (IL6) cytokine family during in vitro bovine oocyte maturation affects maturation success, cumulus–oocyte complex (COC) gene expression, fertilization success, and embryo development potential. Human recombinant proteins for IL6, IL11, and leukemia inhibitory factor (LIF) were supplemented to COCs during the maturation period, then fertilization and embryo culture commenced without further cytokine supplementation. The first study determined that none of these cytokines influenced the rate that oocytes achieved arrest at meiosis II. The second study identified that LIF and IL11 supplementation increases AREG transcript abundance. Supplementation with IL6 supplementation did not affect AREG abundance but reduced HAS2 transcript abundance. Several other transcriptional markers of oocyte competency were not affected by any of the cytokines. The third study determined that supplementing these cytokines during maturation did not influence fertilization success, but either LIF or IL11 supplementation increased blastocyst development. No effect of IL6 supplementation on subsequent blastocyst development was detected. The fourth experiment explored whether each cytokine treatment affects the post-thaw survivability of cryopreserved IVP blastocysts. None of the cytokines supplemented during oocyte maturation produced any positive effects on post-thaw blastocyst re-expansion and hatching. In conclusion, these outcomes implicate IL11 and LIF as potentially useful supplements for improving bovine oocyte competency.
  • An Overview of Reactive Oxygen Species Damage Occurring during In Vitro Bovine Oocyte and Embryo Development and the Efficacy of Antioxidant Use to Limit These Adverse Effects
    Keane, Jessica A.; Ealy, Alan D. (MDPI, 2024-01-21)
    The in vitro production (IVP) of bovine embryos has gained popularity worldwide and in recent years and its use for producing embryos from genetically elite heifers and cows has surpassed the use of conventional superovulation-based embryo production schemes. There are, however, several issues with the IVP of embryos that remain unresolved. One limitation of special concern is the low efficiency of the IVP of embryos. Exposure to reactive oxygen species (ROS) is one reason why the production of embryos with IVP is diminished. These highly reactive molecules are generated in small amounts through normal cellular metabolism, but their abundances increase in embryo culture because of oocyte and embryo exposure to temperature fluctuations, light exposure, pH changes, atmospheric oxygen tension, suboptimal culture media formulations, and cryopreservation. When uncontrolled, ROS produce detrimental effects on the structure and function of genomic and mitochondrial DNA, alter DNA methylation, increase lipid membrane damage, and modify protein activity. Several intrinsic enzymatic pathways control ROS abundance and damage, and antioxidants react with and reduce the reactive potential of ROS. This review will focus on exploring the efficiency of supplementing several of these antioxidant molecules on oocyte maturation, sperm viability, fertilization, and embryo culture.
  • A Multi-Institutional Description of Processes and Outcomes of Postbaccalaureate Research Education Programs in the Mid-Atlantic Region
    Wright, Cynthia F.; Kasman, Laura M.; Robinson, Donita L.; Carey, Gregory B.; Hall, Joshua D.; Lloyd, Joyce A.; Shiang, Rita; Smith, Edward J.; Wilson, Katherine L. (American Association of Medical Colleges, 2024-01-13)
    Outcome data from 6 National Institutes of Health–funded Postbaccalaureate Research Education Programs (PREPs) in the Mid-Atlantic region were combined to give a multi-institutional perspective on their scholars’ characteristics and progress through biomedical research training. The institutions hosting these programs were Johns Hopkins University School of Medicine, the Medical University of South Carolina, the University of Maryland School of Medicine, the University of North Carolina at Chapel Hill, Virginia Commonwealth University, and Virginia Polytechnic Institute and State University. The authors summarize the institutional pathways, demographics, undergraduate institutions, and graduate institutions for a total of 384 PREP scholars who completed the programs by June 2021. A total of 228 (59.3%) of these PREP scholars identified as Black or African American, 116 (30.2%) as Hispanic or Latinx, and 269 (70.0%) as female. The authors found that 376 of 384 scholars (97.9%) who started PREP finished their program, 319 of 376 (84.8%) who finished PREP matriculated into PhD or MD/PhD programs, and 284 of 319 (89.0%) who matriculated have obtained their PhD or are successfully making progress toward their PhD.
  • Editorial: Bio-accessibility of functional compounds and nutrients of animal diets
    Rossi, Luciana; Theodorou, Georgios; Osorio, Johan; Castiglioni, Bianca (Frontiers, 2023-11-16)
  • Reducing barking in a Brazilian animal shelter: A practical intervention
    Baldan, Ana Lucia; Ferreira, Bruna Lima; Warisaia, Vinicius; Feuerbacher, Erica N.; Monticelli, Patricia Ferreira; Gunter, Lisa M. (Elsevier, 2023-08)
    Barking is a common problem in animal shelters. Loud noise is an irritant and stressful to both humans and other animals. In the present study, we tested a positive reinforcement intervention using food delivery with 70 dogs at a municipal animal shelter in Pirassununga, state of São Paulo, Brazil. The experiment consisted of three conditions with three daily phases: pre-intervention (A1), intervention (B), and post-intervention (A2). The intervention consisted of the experimenter (ALB) entering the building in which dogs were housed, stopping at each of its 12 kennels (between 4 and 6 dogs in each kennel), and delivering food to the dogs after they had ceased barking. After the first study condition with its single experimenter, we conducted two other conditions to test the generalization of the intervention with novel stimuli. In Condition 2, the experimenter was accompanied by a student; and in Condition 3, the experimenter was with the same student and a shelter employee. Continuous sound levels (Leq dB) and duration of barking were measured pre- and post-intervention throughout the study's three conditions as well as the amount of time needed to carry out the intervention each day. We found that, on average, both Leq dB and barking duration reduced following the intervention with a decrease in both measures from the beginning to the end of the study. Furthermore, intervention implementation time shortened across the study's conditions, with less than three minutes needed for the intervention to be carried out in Condition 3. In total, our findings suggest that the Barking Reduction Protocol (BRP) is an effective, low-effort intervention that reduces dog barking in the animal shelter. When considering the many issues that compromise the daily lives of shelter dogs, this intervention may be a useful tool in changing dogs’ barking behavior in response to people and improve their welfare as they await adoption.
  • Phosphorylation of RPT6 Controls Its Ability to Bind DNA and Regulate Gene Expression in the Hippocampus of Male Rats during Memory Formation
    Farrell, Kayla; Auerbach, Aubrey; Musaus, Madeline; Navabpour, Shaghayegh; Liu, Catherine; Lin, Yu; Xie, Hehuang; Jarome, Timothy J. (Society for Neuroscience, 2024-01)
    Memory formation requires coordinated control of gene expression, protein synthesis, and ubiquitin–proteasome system (UPS)-mediated protein degradation. The catalytic component of the UPS, the 26S proteasome, contains a 20S catalytic core surrounded by two 19S regulatory caps, and phosphorylation of the 19S cap regulatory subunit RPT6 at serine 120 (pRPT6-S120) has been widely implicated in controlling activity-dependent increases in proteasome activity. Recently, RPT6 was also shown to act outside the proteasome where it has a transcription factor-like role in the hippocampus during memory formation. However, little is known about the proteasome-independent function of “free” RPT6 in the brain or during memory formation and whether phosphorylation of S120 is required for this transcriptional control function. Here, we used RNA-sequencing along with novel genetic approaches and biochemical, molecular, and behavioral assays to test the hypothesis that pRPT6-S120 functions independently of the proteasome to bind DNA and regulate gene expression during memory formation. RNA-sequencing following siRNA-mediated knockdown of free RPT6 revealed 46 gene targets in the dorsal hippocampus of male rats following fear conditioning, where RPT6 was involved in transcriptional activation and repression. Through CRISPR-dCas9-mediated artificial placement of RPT6 at a target gene, we found that RPT6 DNA binding alone may be important for altering gene expression following learning. Further, CRISPR-dCas13-mediated conversion of S120 to glycine on RPT6 revealed that phosphorylation at S120 is necessary for RPT6 to bind DNA and properly regulate transcription during memory formation. Together, we reveal a novel function for phosphorylation of RPT6 in controlling gene transcription during memory formation.
  • Ablation of OCT4 function in cattle embryos by double electroporation of CRISPR-Cas for DNA and RNA targeting (CRISPR-DART)
    Nix, Jada L.; Schettini, Gustavo P.; Speckhart, Savannah L.; Ealy, Alan D.; Biase, Fernando H. (Oxford University Press, 2023-11-01)
    CRISPR-Cas ribonucleoproteins (RNPs) are important tools for gene editing in preimplantation embryos. However, the inefficient production of biallelic deletions in cattle zygotes has hindered mechanistic studies of gene function. In addition, the presence of maternal RNAs that support embryo development until embryonic genome activation may cause confounding phenotypes. Here, we aimed to improve the efficiency of biallelic deletions and deplete specific maternal RNAs in cattle zygotes using CRISPR-Cas editing technology. Two electroporation sessions with Cas9D10A RNPs targeting exon 1 and the promoter of OCT4 produced biallelic deletions in 91% of the embryos tested. In most cases, the deletions were longer than 1,000 nucleotides long. Electroporation of Cas13a RNPs prevents the production of the corresponding proteins. We electroporated Cas9D10A RNPs targeting exon 1, including the promoter region, of OCT4 in two sessions with inclusion of Cas13a RNPs targeting OCT4 mRNAs in the second session to ablate OCT4 function in cattle embryos. A lack of OCT4 resulted in embryos arresting development prior to blastocyst formation at a greater proportion (13%) than controls (31.6%, P < 0.001). The few embryos that developed past the morula stage did not form a normal inner cell mass. Transcriptome analysis of single blastocysts, confirmed to lack exon 1 and promoter region of OCT4, revealed a significant (False Discovery Rate, FDR < 0.1) reduction in transcript abundance of many genes functionally connected to stemness, including markers of pluripotency (CADHD1, DPPA4, GNL3, RRM2). The results confirm that OCT4 is a key regulator of genes that modulate pluripotency and is required to form a functional blastocyst in cattle.
  • Sex linked behavioral and hippocampal transcriptomic changes in mice with cell-type specific Egr1 loss
    Swilley, Cody; Lin, Yu; Zheng, Yuze; Xu, Xiguang; Liu, Min; Jarome, Timothy J.; Hodes, Georgia E.; Xie, Hehuang (Frontiers, 2023-10-19)
    The transcription factor EGR1 is instrumental in numerous neurological processes, encompassing learning and memory as well as the reaction to stress. Egr1 complete knockout mice demonstrate decreased depressive or anxiety-like behavior and impaired performance in spatial learning and memory. Nevertheless, the specific functions of Egr1 in distinct cell types have been largely underexplored. In this study, we cataloged the behavioral and transcriptomic character of Nestin-Cre mediated Egr1 conditional knockout (Egr1cKO) mice together with their controls. Although the conditional knockout did not change nociceptive or anxiety responses, it triggered changes in female exploratory activity during anxiety testing. Hippocampus-dependent spatial learning in the object location task was unaffected, but female Egr1cKO mice did exhibit poorer retention during testing on a contextual fear conditioning task compared to males. RNA-seq data analyses revealed that the presence of the floxed Egr1 cassette or Nestin-Cre driver alone exerts a subtle influence on hippocampal gene expression. The sex-related differences were amplified in Nestin-Cre mediated Egr1 conditional knockout mice and female mice are more sensitive to the loss of Egr1 gene. Differentially expressed genes resulted from the loss of Egr1 in neuronal cell lineage were significantly associated with the regulation of Wnt signaling pathway, extracellular matrix, and axon guidance. Altogether, our results demonstrate that Nestin-Cre and the loss of Egr1 in neuronal cell lineage have distinct impacts on hippocampal gene expression in a sex-specific manner.
  • Comparison of Methods for Individual Killing of Broiler Chickens: A Matter of Animal Welfare and On-Farm Feasibility
    Watteyn, Anneleen; Garmyn, An; Ampe, Bart; Jacobs, Leonie; Moons, Christel P. H.; Tuyttens, Frank A. M. (Frontiers, 2022-05-30)
    The humane killing of individual broiler chickens on-farm requires a minimum of suffering. In this regard, rapid and irreversible loss of consciousness are important determinants. This can be verified by cerebral and spinal reflexes. Also, on-farm feasibility determines whether producers will apply the method. The aim of the study was to compare the effectiveness and animal welfare impact of two different methods for killing individual broilers of varying ages (2, 4, and 6 weeks): manual cervical dislocation (CD) and captive bolt (CB). The evaluation of CD and CB was based on effectiveness and on time to onset (convulsions) or cessation (pain response, pupillary light reflex, convulsions, heartbeat) of non-invasive indicators. In addition, a pilot study was conducted on-farm to assess the feasibility of two alternative methods, CB and nitrogen gasification (N2), and to survey farmers’ opinions on them. The onset of convulsions was almost immediate for both methods in the first study. No differences between CD and CB were observed for the cessation of pain response for chickens at age of 2 weeks (5.0 and 7.5 s, respectively) and 6 weeks (14.0 and 14.1 s, respectively). However, at 4 weeks a longer pain response was measured after CD (11.3 s) than after CB (4.7 s). For the three age categories, the pupillary light reflex disappeared later after CD (54.9 - 80.7 s) compared to CB (8.3 - 13.7 s). The same was observed for cessation of convulsions in 2- and 6-week-old chickens (185.3 and 172.0 s for CD and 79.0 and 82.9 s for CB). This suggests that brain death occurred faster after CB compared to CD. No difference between the methods was found for the cessation of the heartbeat. After the pilot study, the producers preferred N2 over CB in terms of animal-friendliness, time-efficiency, ease of use, and effectiveness. However, both methods were found rather expensive and required some experience. CB and N2 are good killing alternatives to CD due to rapid and irreversible insensibility. However, more information and support for chicken producers will be needed for these to become routine killing methods.
  • Genotype-by-environment interactions for feed efficiency traits in Nellore cattle based on bi-trait reaction norm models
    Silva Neto, João B.; Mota, Lucio F. M.; Amorim, Sabrina T.; Peripolli, Elisa; Brito, Luiz F.; Magnabosco, Claudio U.; Baldi, Fernando (2023-12-14)
    Background: Selecting animals for feed efficiency directly impacts the profitability of the beef cattle industry, which contributes to minimizing the environmental footprint of beef production. Genetic and environmental factors influence animal feed efficiency, leading to phenotypic variability when exposed to different environmental conditions (i.e., temperature and nutritional level). Thus, our aim was to assess potential genotype-by-environment (G × E) interactions for dry matter intake (DMI) and residual feed intake (RFI) in Nellore cattle (Bos taurus indicus) based on bi-trait reaction norm models (RN) and evaluate the genetic association between RFI and DMI across different environmental gradient (EG) levels. For this, we used phenotypic information on 12,958 animals (young bulls and heifers) for DMI and RFI recorded during 158 feed efficiency trials. Results: The heritability estimates for DMI and RFI across EG ranged from 0.26 to 0.54 and from 0.07 to 0.41, respectively. The average genetic correlations (± standard deviation) across EG for DMI and RFI were 0.83 ± 0.19 and 0.81 ± 0.21, respectively, with the lowest genetic correlation estimates observed between extreme EG levels (low vs. high) i.e. 0.22 for RFI and 0.26 for DMI, indicating the presence of G × E interactions. The genetic correlation between RFI and DMI across EG levels decreased as the EG became more favorable and ranged from 0.79 (lowest EG) to 0.52 (highest EG). Based on the estimated breeding values from extreme EG levels (low vs. high), we observed a moderate Spearman correlation of 0.61 (RFI) and 0.55 (DMI) and a selection coincidence of 53.3% and 40.0% for RFI and DMI, respectively. Conclusions: Our results show evidence of G × E interactions on feed efficiency traits in Nellore cattle, especially in feeding trials with an average daily gain (ADG) that is far from the expected of 1 kg/day, thus increasing reranking of animals.
  • ARGem: a new metagenomics pipeline for antibiotic resistance genes: metadata, analysis, and visualization
    Liang, Xiao; Zhang, Jingyi; Kim, Yoonjin; Ho, Josh; Liu, Kevin; Keenum, Ishi M.; Gupta, Suraj; Davis, Benjamin; Hepp, Shannon L.; Zhang, Liqing; Xia, Kang; Knowlton, Katharine F.; Liao, Jingqiu; Vikesland, Peter J.; Pruden, Amy; Heath, Lenwood S. (Frontiers, 2023-09-15)
    Antibiotic resistance is of crucial interest to both human and animal medicine. It has been recognized that increased environmental monitoring of antibiotic resistance is needed. Metagenomic DNA sequencing is becoming an attractive method to profile antibiotic resistance genes (ARGs), including a special focus on pathogens. A number of computational pipelines are available and under development to support environmental ARG monitoring; the pipeline we present here is promising for general adoption for the purpose of harmonized global monitoring. Specifically, ARGem is a user-friendly pipeline that provides full-service analysis, from the initial DNA short reads to the final visualization of results. The capture of extensive metadata is also facilitated to support comparability across projects and broader monitoring goals. The ARGem pipeline offers efficient analysis of a modest number of samples along with affordable computational components, though the throughput could be increased through cloud resources, based on the user’s configuration. The pipeline components were carefully assessed and selected to satisfy tradeoffs, balancing efficiency and flexibility. It was essential to provide a step to perform short read assembly in a reasonable time frame to ensure accurate annotation of identified ARGs. Comprehensive ARG and mobile genetic element databases are included in ARGem for annotation support. ARGem further includes an expandable set of analysis tools that include statistical and network analysis and supports various useful visualization techniques, including Cytoscape visualization of co-occurrence and correlation networks. The performance and flexibility of the ARGem pipeline is demonstrated with analysis of aquatic metagenomes. The pipeline is freely available at https://github.com/xlxlxlx/ARGem.
  • The Influence of Brief Outing and Temporary Fostering Programs on Shelter Dog Welfare
    Gunter, Lisa M.; Blade, Emily M.; Gilchrist, Rachel J.; Nixon, Betsy J.; Reed, Jenifer L.; Platzer, Joanna M.; Wurpts, Ingrid C.; Feuerbacher, Erica N.; Wynne, Clive D. L. (MDPI, 2023-11-15)
    Human interaction is one of the most consistently effective interventions that can improve the welfare of shelter-living dogs. Time out of the kennel with a person has been shown to reduce physiological measures of stress as can leaving the shelter for a night or more in a foster home. In this study, we assessed the effects of brief outings and temporary fostering stays on dogs’ length of stay and outcomes. In total, we analyzed data of 1955 dogs from 51 animal shelters that received these interventions as well as 25,946 dogs residing at these shelters that served as our controls. We found that brief outings and temporary fostering stays increased dogs’ likelihood of adoption by 5.0 and 14.3 times, respectively. While their lengths of stay were longer in comparison to control dogs, this difference was present prior to the intervention. Additionally, we found that these programs were more successful when greater percentages of community members (as compared to volunteers and staff) were involved in caregiving as well as when programs were implemented by better-resourced shelters. As such, animal welfare organizations should consider implementing these fostering programs as evidence-based best practices that can positively impact the outcomes of shelter dogs.
  • Review—Prospects in Cancer Diagnosis: Exosome-Chip for Liquid Biopsy
    Khondakar, Kamil Reza; Ataei Kachouei, Matin; Erukainure, Frank Efe; Ali, Md Azahar (The Electrochemical Society, 2023-12-01)
    A liquid biopsy combined with an exosome-chip (EC) is an important detection tool for early cancer diagnosis. Exosomes have a crucial function in the exchange of information between cells and are present in biological fluids. ECs are miniaturized microfluidic devices designed to isolate, capture, and analyze exosomes for analysis of patient samples. Such devices offer on-chip detection, high-throughput analysis, and multiplex measurements. Further, these chips can integrate with electrochemical and optical detectors, and mass spectrometry enabling comprehensive studies of diseases. This review will cover the outlook on chip-based diagnostics for liquid biopsy, detection, and isolation of exosomes to support cancer diagnostics.
  • Sex-differences in proteasome-dependent K48-polyubiquitin signaling in the amygdala are developmentally regulated in rats
    Farrell, Kayla; Auerbach, Aubrey; Liu, Catherine; Martin, Kiley; Pareno, Myasia; Ray, W. Keith; Helm, Richard F.; Biase, Fernando; Jarome, Timothy J. (2023-11-10)
    Background Sex differences have been observed in several brain regions for the molecular mechanisms involved in baseline (resting) and memory-related processes. The ubiquitin proteasome system (UPS) is a major protein degradation pathway in cells. Sex differences have been observed in lysine-48 (K48)-polyubiquitination, the canonical degradation mark of the UPS, both at baseline and during fear memory formation within the amygdala. Here, we investigated when, how, and why these baseline sex differences arise and whether both sexes require the K48-polyubiquitin mark for memory formation in the amygdala. Methods We used a combination of molecular, biochemical and proteomic approaches to examine global and protein-specific K48-polyubiquitination and DNA methylation levels at a major ubiquitin coding gene (Uba52) at baseline in the amygdala of male and female rats before and after puberty to determine if sex differences were developmentally regulated. We then used behavioral and genetic approaches to test the necessity of K48-polyubiquitination in the amygdala for fear memory formation. Results We observed developmentally regulated baseline differences in Uba52 methylation and total K48-polyubiquitination, with sexual maturity altering levels specifically in female rats. K48-polyubiquitination at specific proteins changed across development in both male and female rats, but sex differences were present regardless of age. Lastly, we found that genetic inhibition of K48-polyubiquitination in the amygdala of female, but not male, rats impaired fear memory formation. Conclusions These results suggest that K48-polyubiquitination differentially targets proteins in the amygdala in a sex-specific manner regardless of age. However, sexual maturity is important in the developmental regulation of K48-polyubiquitination levels in female rats. Consistent with these data, K48-polyubiquitin signaling in the amygdala is selectively required to form fear memories in female rats. Together, these data indicate that sex-differences in baseline K48-polyubiquitination within the amygdala are developmentally regulated, which could have important implications for better understanding sex-differences in molecular mechanisms involved in processes relevant to anxiety-related disorders such as post-traumatic stress disorder (PTSD).
  • Fibroblast Growth Factor 21 Has a Diverse Role in Energetic and Reproductive Physiological Functions of Female Beef Cattle
    Prezotto, Ligia D.; Keane, Jessica A.; Cupp, Andrea S.; Thorson, Jennifer F. (MDPI, 2023-10-12)
    Fibroblast growth factor 21 (FGF21) has been identified in multiple mammalian species as a molecular marker of energy metabolism while also providing negative feedback to the gonads. However, the role of FGF21 in regulating the energetic and reproductive physiology of beef heifers and cows has yet to be characterized. Herein, we investigated the temporal concentrations of FGF21 in female beef cattle from the prepubertal period to early lactation. Circulating concentrations of FGF21, non-esterified fatty acids, plasma urea nitrogen, glucose, and progesterone were assessed. Ultrasonography was employed to determine the onset of puberty and resumption of postpartum ovarian cyclicity as well as to measure backfat thickness. Finally, cows and calves underwent the weigh-suckle-weigh technique to estimate rate of milk production. We have revealed that FGF21 has an expansive role in the physiology of female beef cattle, including pubertal onset, adaptation to nutritional transition, rate of body weight gain, circulating markers of metabolism, and rate of milk production. In conclusion, FGF21 plays a role in physiological functions in beef cattle that can be applied to advance the understanding of basic scientific processes governing the nutritional regulation of reproductive function but also provides a novel means for beef cattle producers to select parameters of financial interest.
  • Profiling intestinal stem and proliferative cells in the small intestine of broiler chickens via in situ hybridization during the peri-hatch period
    Cloft, Sara E.; Uni, Zehava; Wong, Eric A. (Elsevier, 2023-04)
    Mature small intestines have crypts populated by stem cells which produce replacement cells to maintain the absorptive villus surface area. The embryonic crypt is rudimentary and cells along the villi are capable of proliferation. By 7 d post-hatch the crypts are developed and are the primary sites of proliferation. Research characterizing the proliferative expansion of the small intestine during the peri-hatch period is lacking. The objective of this study was to profile the changes of genes that are markers of stem cells and proliferation: Olfactomedin 4 (Olfm4), Leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5), and marker of proliferation Ki67 from embryonic day 17 to 7 d post-hatch using quantitative PCR and in situ hybridization (ISH). The expression of the stem cell marker genes differed. Olfm4 mRNA increased while Lgr5 mRNA decreased post-hatch. Ki67 mRNA decreased post-hatch in the duodenum and was generally the greatest in the ileum. The ISH was consistent with the quantitative PCR results. Olfm4 mRNA was only seen in the crypts and increased with morphological development of the crypts. In contrast Lgr5 mRNA was expressed in the crypt and the villi in the embryonic periods but became restricted to the intestinal crypt during the post-hatch period. Ki67 mRNA was expressed throughout the intestine pre-hatch, but then expression became restricted to the crypt and the center of the villi. The ontogeny of Olfm4, Lgr5, and Ki67 expressing cells show that proliferation in the peri-hatch intestine changes from along the entire villi to being restricted within the crypts.
  • Testing for BLV: Control, reduction, and elimination?
    Corl, Benjamin A.; Ali, Md. Azahar (Virginia State Dairymen's Association, 2023-10-01)