Integrated analyses using medaka as a powerful model animal toward understanding various aspects of reproductive regulation

Glucuronides hydrolysis by intestinal microbial β-Glucuronidases (GUS) is an important procedure for many endogenous and exogenous compounds. The purpose of this study is to determine the impact of experimental conditions on glucuronide hydrolysis by intestinal microbial GUS. Standard probe 4-Nitrophenyl β-D-glucopyranoside (pNPG) and a natural glucuronide wogonoside were used as the model compounds. Feces collection time, buffer conditions, interindividual, and species variations were evaluated by incubating the substrates with enzymes. The relative reaction activity of pNPG, reaction rates, and reaction kinetics for wogonoside were calculated. Fresh feces showed the highest hydrolysis activities. Sonication increased total protein yield during enzyme preparation. The pH of the reaction system increased the activity in 0.69–1.32-fold, 2.9–12.9-fold, and 0.28–1.56-fold for mouse, rat, and human at three different concentrations of wogonoside, respectively. The Vmax for wogonoside hydrolysis was 2.37 ± 0.06, 4.48 ± 0.11, and 5.17 ± 0.16 μmol/min/mg and Km was 6.51 ± 0.71, 3.04 ± 0.34, and 0.34 ± 0.047 μM for mouse, rat, and human, respectively. The inter-individual difference was significant (4–6-fold) using inbred rats as the model animal. Fresh feces should be used to avoid activity loss and sonication should be utilized in enzyme preparation to increase hydrolysis activity. The buffer pH should be appropriate according to the species. Inter-individual and species variations were significant.

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Seminal Plasma: Relevant for Fertility?

International Journal of Molecular Sciences ◽

10.3390/ijms22094368 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4368

Author(s):

Heriberto Rodriguez-Martinez ◽

Emilio A. Martinez ◽

Juan J. Calvete ◽

Fernando J. Peña Vega ◽

Jordi Roca

Keyword(s):

Seminal Plasma ◽

Current Knowledge ◽

Inorganic Ions ◽

Vitro Fertilization ◽

Dna And Rna ◽

Model Animal ◽

Sexual Glands ◽

Species Specific

Seminal plasma (SP), the non-cellular component of semen, is a heterogeneous composite fluid built by secretions of the testis, the epididymis and the accessory sexual glands. Its composition, despite species-specific anatomical peculiarities, consistently contains inorganic ions, specific hormones, proteins and peptides, including cytokines and enzymes, cholesterol, DNA and RNA—the latter often protected within epididymis- or prostate-derived extracellular vesicles. It is beyond question that the SP participates in diverse aspects of sperm function pre-fertilization events. The SP also interacts with the various compartments of the tubular genital tract, triggering changes in gene function that prepares for an eventual successful pregnancy; thus, it ultimately modulates fertility. Despite these concepts, it is imperative to remember that SP-free spermatozoa (epididymal or washed ejaculated) are still fertile, so this review shall focus on the differences between the in vivo roles of the SP following semen deposition in the female and those regarding additions of SP on spermatozoa handled for artificial reproduction, including cryopreservation, from artificial insemination to in vitro fertilization. This review attempts, including our own results on model animal species, to critically summarize the current knowledge of the reproductive roles played by SP components, particularly in our own species, which is increasingly affected by infertility. The ultimate goal is to reconcile the delicate balance between the SP molecular concentration and their concerted effects after temporal exposure in vivo. We aim to appraise the functions of the SP components, their relevance as diagnostic biomarkers and their value as eventual additives to refine reproductive strategies, including biotechnologies, in livestock models and humans.

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In Search of Species-Specific SNPs in a Non-Model Animal (European Bison (Bison bonasus))—Comparison of De Novo and Reference-Based Integrated Pipeline of STACKS Using Genotyping-by-Sequencing (GBS) Data

Animals ◽

10.3390/ani11082226 ◽

2021 ◽

Vol 11 (8) ◽

pp. 2226

Author(s):

Sazia Kunvar ◽

Sylwia Czarnomska ◽

Cino Pertoldi ◽

Małgorzata Tokarska

Keyword(s):

Reference Genome ◽

De Novo ◽

Bos Taurus ◽

Model Organism ◽

Genotyping By Sequencing ◽

Model Organisms ◽

European Bison ◽

Model Animal ◽

Pcr Duplicates ◽

Species Specific

The European bison is a non-model organism; thus, most of its genetic and genomic analyses have been performed using cattle-specific resources, such as BovineSNP50 BeadChip or Illumina Bovine 800 K HD Bead Chip. The problem with non-specific tools is the potential loss of evolutionary diversified information (ascertainment bias) and species-specific markers. Here, we have used a genotyping-by-sequencing (GBS) approach for genotyping 256 samples from the European bison population in Bialowieza Forest (Poland) and performed an analysis using two integrated pipelines of the STACKS software: one is de novo (without reference genome) and the other is a reference pipeline (with reference genome). Moreover, we used a reference pipeline with two different genomes, i.e., Bos taurus and European bison. Genotyping by sequencing (GBS) is a useful tool for SNP genotyping in non-model organisms due to its cost effectiveness. Our results support GBS with a reference pipeline without PCR duplicates as a powerful approach for studying the population structure and genotyping data of non-model organisms. We found more polymorphic markers in the reference pipeline in comparison to the de novo pipeline. The decreased number of SNPs from the de novo pipeline could be due to the extremely low level of heterozygosity in European bison. It has been confirmed that all the de novo/Bos taurus and Bos taurus reference pipeline obtained SNPs were unique and not included in 800 K BovineHD BeadChip.

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Nucleated red blood cells participate in myocardial regeneration in the toad Bufo Gargarizan Gargarizan

Experimental Biology and Medicine ◽

10.1177/15353702211013297 ◽

2021 ◽

pp. 153537022110132

Author(s):

Shu-Qin Liu ◽

Xiao-Ye Hou ◽

Feng Zhao ◽

Xiao-Ge Zhao

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Cardiac Injury ◽

Myocardial Regeneration ◽

Matrix Metalloproteinase 2 ◽

Enzyme Linked Immunosorbent Assay ◽

Cardiac Tissues ◽

Nucleated Red Blood Cells ◽

Model Animal

Heart regeneration is negligible in humans and mammals but remarkable in some ectotherms. Humans and mammals lack nucleated red blood cells (NRBCs), while ectotherms have sufficient NRBCs. This study used Bufo gargarizan gargarizan, a Chinese toad subspecies, as a model animal to verify our hypothesis that NRBCs participate in myocardial regeneration. NRBC infiltration into myocardium was seen in the healthy toad hearts. Heart needle-injury was used as an enlarged model of physiological cardiomyocyte loss. It recovered quickly and scarlessly. NRBC infiltration increased during the recovery. Transwell assay was done to in vitro explore effects of myocardial injury on NRBCs. In the transwell system, NRBCs could infiltrate into cardiac pieces and could transdifferentiate toward cardiomyocytes. Heart apex cautery caused approximately 5% of the ventricle to be injured to varying degrees. In the mildly to moderately injured regions, NRBC infiltration increased and myocardial regeneration started soon after the inflammatory response; the severely damaged region underwent inflammation, scarring, and vascularity before NRBC infiltration and myocardial regeneration, and recovered scarlessly in four months. NRBCs were seen in the newly formed myocardium. Enzyme-linked immunosorbent assay and Western blotting showed that the levels of tumor necrosis factor-α, interleukin- 1β, 6, and11, cardiotrophin-1, vascular endothelial growth factor, erythropoietin, matrix metalloproteinase- 2 and 9 in the serum and/or cardiac tissues fluctuated in different patterns during the cardiac injury-regeneration. Cardiotrophin-1 could induce toad NRBC transdifferentiation toward cardiomyocytes in vitro. Taken together, the results suggest that the NRBC is a cell source for cardiomyocyte renewal/regeneration in the toad; cardiomyocyte loss triggers a series of biological processes, facilitating NRBC infiltration and transition to cardiomyocytes. This finding may guide a new direction for improving human myocardial regeneration.

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Relation between Feed Particle Size Distribution and Plumage Condition in Laying Hens on Commercial Farms

Animals ◽

10.3390/ani11030773 ◽

2021 ◽

Vol 11 (3) ◽

pp. 773

Author(s):

Ruben Schreiter ◽

Klaus Damme ◽

Markus Freick

Keyword(s):

Damage Control ◽

Geometric Mean ◽

Cross Sectional Study ◽

Cross Sectional ◽

Nutrient Analysis ◽

Indirect Measure ◽

Model Animal ◽

Commercial Farms ◽

And Control ◽

Feed Samples

In this cross-sectional study, 103 complete feed samples from laying hen herds affected by plumage damage as an indirect measure for severe feather pecking (affected herds; AH, n = 37) and control herds without plumage damage (control herd; CH, n = 66) of commercial German farms were examined by dry sieve and nutrient analysis. AH showed higher percentages of particles >2.50 mm (mean ± SD, CH: 11.0 ± 8.5%, AH: 24.9 ± 14.3%) and 2.00–2.50 mm (CH: 11.2 ± 5.3%, AH: 15.7 ± 5.7%), but lower proportions of fractions 1.01–1.60 mm (CH: 22.9 ± 4.9%, AH: 17.8 ± 5.7%), 0.51–1.00 mm (CH: 25.5 ± 8.2%, AH: 16.0 ± 6.8%) and ≤0.50 mm (CH: 15.4 ± 5.0%, AH: 11.0 ± 4.8%) (p < 0.001). Diets of AH had a higher geometric mean diameter (GMD) compared to CH (AH: 1470.8 ± 343.9 μm; CH: 1113.3 ± 225.7 μm) (p < 0.001). Contents of crude ash (CH: 130.3 ± 18.8 g/kg, AH: 115.9 ± 24.3 g/kg), lysine (CH: 8.2 ± 1.0 g/kg, AH: 7.7 ± 1.2 g/kg), methionine (CH: 3.4 ± 0.5 g/kg, AH: 3.2 ± 0.6 g/kg) and sodium (CH: 1.7 ± 0.4 g/kg, AH: 1.3 ± 0.4 g/kg) were lower in AH (p ≤ 0.041). In a logistic regression model, animal age (p = 0.041) and GMD (p < 0.001) were significant factors on the occurrence of plumage damage.

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