Hormonal biochemical monitoring at bull-producers of different breeds in the conditions of the Central Black Economic District

Purpose: To conduct a comprehensive monitoring of bulls of different breeds in the summer in a comparative perspective to the study of protein-lipid and mineral metabolism, as well as to determine hormonal status on the content of endogenous hormones: testosterone, estradiol, cortisol and serum thyroxine per day taking seed.Materials and methods. For the first time in a Chernozem region of the Russian Federation (Voronezh region) was carried out comprehensive monitoring of bulls of red-motley Holstein (n = 16), Red-and-White (n = 8) and Simmental (n = 6) rocks in the same time of year (summer) of the protein-lipid and mineral metabolism, as well as the level of endogenous hormone (testosterone, estradiol, cortisol, thyroxine) and their cholesterol precursor in comparative aspect, immediately after taking the seed. Age range bulls ranged from 2 to 11 years. General monitoring was conducted on 23 indices of blood serum 3 rocks. The studies were conducted by modern methods, using modern equipment and reagents.Results. At Red Pied Holsteins an increase of total protein 100.45 g / L versus 92 g / l in normal (above the norm of 9.2%, as compared with red and White breed at the level of 11.3%). A similar increase in concentration in the blood of Holstein bulls red-White breed recorded on globulins, 68.87 g / l to 63 g / l in normal (above the norm of 9.3%), and with respect to the red-and-White breed 17, 6%. Also notes the high content of AST bulls red-motley Holstein against the red-and-White 18%, but within the reference range. Condition protein and lipid metabolism in sires only differed in content of creatinine, which amounted to 165.04 micromol / l against 163 mmol / l in normal, 1.6% higher as compared to the red and white breed this index was higher 18.7%. According to the content of testosterone highest rate was observed in bulls red-White breed 54.73 ± 9.51 nmol / l, is 10.2% more than in bulls red checkered-Holstein and 12.45% compared with Simmental breed. Similarly, in terms of serum cortisol - 355.60 ± 39.2 nmol / l in the red-bulls White breed that more than 2-fold compared with bulls Red Pied Holstein and Simmental. The difference is statistically significant at P ≤ 0,01. According to the content of estradiol relatively low levels recorded in the bulls of red-motley breed - 0.265 nmol / L vs. 0.301 nmol / L in red bulls-motley Holstein, and 0.332 nmol / L - Simmental, is lower by 13.6% and 25 , 3%, respectively.Conclusion. It was found that substantially all of the studied parameters in bulls 3 breeds are at reference values, and this gives grounds to assume that all of these breeds are well adapted to the conditions of Black Soil.

Hormonal state of breeding bulls after a long winter period of operation in the conditions of the Udmurt Republic

Relevance. The study will help to identify new perspectives in breeding; get new data about the biological processes occurring in the body of animals according to the endogenous hormones of breeding bulls. The hormonal-immunobiological relationship in the body of bulls and their adaptive ability in different climatic conditions is a very important issue today.Methods. The work was performed in a temperate continental climate with a long, snowy winter and warm summer in the conditions of the Udmurt Republic in the period from 2020 to 2021 on breeding bulls (n = 27) of modern breeding at the age of 15–69 months. The state of endogenous hormones testosterone, estradiol, thyroxine and cortisol after a long winter period was studied, depending on the age, breed and selection in the normal physiological functioning of breeding bulls. The average serum values of estradiol — 0.517 nmol/l, testosterone — 25.7 nmol/l, thyroxine — 74.7 nmol/l, cortisol — 420 nmol/l, and their cholesterol precursor at the level of 3.2 nmol/l were established in the blood of breeding bulls located in this region. It was found that, depending on age, there are differences in all the studied indicators, however, there is a reliability for thyroxine and cortisol. The concentration of cortisol by selection significantly differs in imported bulls of European selection from the Netherlands at the level of p > 0.05, which characterizes the presence of a certain stress adaptation factor in this group. In terms of variability, a significant difference between min and max indicators was noted for the concentration of testosterone, which varied between 2.4 nmol/l and 60 nmol/l, indicating the individuality of these hormones in breeding bulls in this region.Results. Based on the data obtained, the need to take these data into account when operating breeding bulls and analyzing sperm products, taking into account the concentration of these hormones, as well as the implementation of appropriate preventive measures, is revealed.

Atxn2-CAG100-KnockIn mouse spinal cord shows progressive TDP43 pathology associated with cholesterol biosynthesis suppression

Large polyglutamine expansions in Ataxin-2 (ATXN2) cause multi-system nervous atrophy in Spinocerebellar Ataxia type 2 (SCA2). Intermediate size expansions carry a risk for selective motor neuron degeneration, known as Amyotrophic Lateral Sclerosis (ALS). Conversely, the depletion of ATXN2 prevents disease progression in ALS. Although ATXN2 interacts directly with RNA, and in ALS pathogenesis there is a crucial role of RNA toxicity, the affected functional pathways remain ill defined. Here, we examined an authentic SCA2 mouse model with Atxn2-CAG100-KnockIn for a first definition of molecular mechanisms in spinal cord pathology. Neurophysiology of lower limbs detected sensory neuropathy rather than motor denervation. Triple immunofluorescence demonstrated cytosolic ATXN2 aggregates sequestrating TDP43 and TIA1 from the nucleus. In immunoblots, this was accompanied by elevated CASP3, RIPK1 and PQBP1 abundance. RT-qPCR showed increase of Grn, Tlr7 and Rnaset2 mRNA versus Eif5a2, Dcp2, Uhmk1 and Kif5a decrease. These SCA2 findings overlap well with known ALS features. Similar to other ataxias and dystonias, decreased mRNA levels for Unc80, Tacr1, Gnal, Ano3, Kcna2, Elovl5 and Cdr1 contrasted with Gpnmb increase. Preterminal stage tissue showed strongly activated microglia containing ATXN2 aggregates, with parallel astrogliosis. Global transcriptome profiles from stages of incipient motor deficit versus preterminal age identified molecules with progressive downregulation, where a cluster of cholesterol biosynthesis enzymes including Dhcr24, Msmo1, Idi1 and Hmgcs1 was prominent. Gas chromatography demonstrated a massive loss of crucial cholesterol precursor metabolites. Overall, the ATXN2 protein aggregation process affects diverse subcellular compartments, in particular stress granules, endoplasmic reticulum and receptor tyrosine kinase signaling. These findings identify new targets and potential biomarkers for neuroprotective therapies.

Novel Approaches for Detection Fluorescent-Labeled by Cellvizio Lab System on Hippocampal CA1 Region

Neurosteroids have been identified in the 1981. Dehydroepiandrosterone sulphate (DHEAS) is one of the vital neurosteroids that de novo synthesized in the nervous system from cholesterol precursor (Baulieu & Robel, 1998). The aim of the study is to develop a method for fluorescence labelling. Alexa Fluor 488 dye with DHEAS antibody can binds the DHEAS antibody in the rat brain monitored by Cellvizio Lab System. DHEAS antibody (IgG isotype antibodies) was fluorescently conjugated by an amine-reactive compound, Alexa Fluor 5-SDP ester 488 dye. The resultant Alexa Fluor 488-conjugated antibodies were collected and analyzed by UV-Vis spectrophotometer instrument. The absorbance of the protein-dye conjugate at 280 nm and 494 nm were measured. Then, the degree of labeling (DOL) was calculated to achieve the desired results. Fluorescence labelling were carried out into the CA1 region of hippocampus Sprague-Dawley rat. We reported that the conjugation was successful. Optimal labeling depending on degree of labeling (DOL) needs some necessity to achieve and effective binding to the target neurosteroid, DHEAS. Cellvizio Lab system connected with Fiber Fluorescence Microscopy (FFM) probe is presented as a new approach in real-time imaging of DHEAS. In conclusion, we have developed a new method of DHEAS-Alexa Fluor fluorescence labelling to visualize and evaluate the changes of DHEAS fluorescence level in the rat hippocampus. This novel approach as a diagnostic tool and can be used to better understand the mechanisms and functions of DHEAS and other neurosteroids in future research.

Patched1 and Patched2 inhibit Smoothened non-cell autonomously

Smoothened (Smo) inhibition by Patched (Ptch) is central to Hedgehog (Hh) signaling. Ptch, a proton driven antiporter, is required for Smo inhibition via an unknown mechanism. Hh ligand binding to Ptch reverses this inhibition and activated Smo initiates the Hh response. To determine whether Ptch inhibits Smo strictly in the same cell or also mediates non-cell-autonomous Smo inhibition, we generated genetically mosaic neuralized embryoid bodies (nEBs) from mouse embryonic stem cells (mESCs). These experiments utilized novel mESC lines in which Ptch1, Ptch2, Smo, Shh and 7dhcr were inactivated via gene editing in multiple combinations, allowing us to measure non-cell autonomous interactions between cells with differing Ptch1/2 status. In several independent assays, the Hh response was repressed by Ptch1/2 in nearby cells. When 7dhcr was targeted, cells displayed elevated non-cell autonomous inhibition. These findings support a model in which Ptch1/2 mediate secretion of a Smo-inhibitory cholesterol precursor.

Apolipoprotein E ( ApoE ) polymorphism is related to differences in potential fertility in women: a case of antagonistic pleiotropy?

The alleles that are detrimental to health, especially in older age, are thought to persist in populations because they also confer some benefits for individuals (through antagonistic pleiotropy). The ApoE4 allele at the ApoE locus, encoding apolipoprotein E (ApoE), significantly increases risk of poor health, and yet it is present in many populations at relatively high frequencies. Why has it not been replaced by natural selection with the health-beneficial ApoE3 allele? ApoE is a major supplier of cholesterol precursor for the production of ovarian oestrogen and progesterone, thus ApoE has been suggested as the potential candidate gene that may cause variation in reproductive performance. Our results support this hypothesis showing that in 117 regularly menstruating women those with genotypes with at least one ApoE4 allele had significantly higher levels of mean luteal progesterone (144.21 pmol l −1 ) than women with genotypes without ApoE4 (120.49 pmol l −1 ), which indicates higher potential fertility. The hormonal profiles were based on daily data for entire menstrual cycles. We suggest that the finding of higher progesterone in women with ApoE4 allele could provide first strong evidence for an evolutionary mechanism of maintaining the ancestral and health-worsening ApoE4 allele in human populations.