Regulation of Folliculogenesis by Growth Factors in Piglet Ovary Exposed Prenatally to β-Hydroxy-β-Methylbutyrate (HMB)

Abstract

<jats:title>Abstract</jats:title><jats:p>Β-hydroxy-β-methylbutyrate (HMB) is one of the leucine metabolites with protein anabolic effects which makes it very popular among athletes. Previously, it was shown that HMB administered during the prenatal period reduced the pool of primordial follicles and increased the proportion of developing follicles in newborn piglets. This work is a further step to understand these morphological alterations. Therefore, the aim of this study was to examine the effect of prenatal HMB treatment on the expression of the Kit ligand, BMP-4, bFGF, and the IGF-1/IGF-1R system which are the main growth factors controlling follicular development. Excised ovaries from 12 newborn piglets, originated from the control (n=6) and HMB-treated (n=6) sows were used for immunohistochemical and western-blot analysis. The tested proteins were localized within egg nests and ovarian follicles. Furthermore, the western-blot assay indicated higher BMP-4, Kit ligand, and IGF-1R expression, while the level of bFGF and IGF-1 proteins decreased after HMB dietary treatment. These findings show that HMB included into sow diet can modulate the expression of growth factors and thereby alter ovarian morphology in offspring. Therefore, this study opens a discussion about the benefits and risks of the diet supplemented with HMB and its potential application in medicine and animal husbandry, and further research is necessary in this area.</jats:p>

Modern Hybrid Rye, as an Alternative Energy Source for Broiler Chickens, Improves the Absorption Surface of the Small Intestine Depending on the Intestinal Part and Xylanase Supplementation

“…Tissue samples obtained from relevant segments, adjacent to the above-mentioned histology samples of the small intestine, were collected from every sacrificed chicken and were used for Western-Blot analysis, as previously described previously [ 26 ]. Briefly, the samples were homogenized in lysis buffer (125 mM TRIS-HCl pH 6.8; 4% SDS; 10% glycerol; 100 mM DTT), boiled in a water bath for 10 min and centrifuged at 10,000× g for 10 min.…”

Femoral µCT Analysis, Mechanical Testing and Immunolocalization of Bone Proteins in β-Hydroxy β-Methylbutyrate (HMB) Supplemented Spiny Mouse in a Model of Pregnancy and Lactation-Associated Osteoporosis

“…As a secondary antibody, a Bright Vision +Poly-HRP-Anti Ms/Rb IgG Biotin-free (Immunologic, Duiven, Netherlands) kit was used, while 3,3′-diaminobenzidine tetrahydrochloride (DAB, D5905, Sigma-Aldrich, St. Louis, MO, USA) was used as a substrate-staining chromogen. Counterstaining was performed with Mayer’s hematoxylin (MHS32-1L, Sigma-Aldrich) [ 42 , 43 ]. Negative control sections for each antibody were conducted by identical immunohistochemical staining excluding the primary antibody ( Supplementary Data, Figure S1 ).…”

Prenatal programming of the small intestine in piglets: the effect of supplementation with 3-hydroxy-3-methylbutyric acid (HMB) in pregnant sows on the structure of jejunum of their offspring

“…Following primary antibodies were used: rabbit polyclonal anti-Ki67 antibody (ab15580); mouse monoclonal anti-neurofilament heavy (poly)peptide antibody to localize myenteric and submucosal ganglia (ab187374); mouse monoclonal anti-active caspase-3 (ab208161); rabbit polyclonal anti-VIP (ab22736), rabbit polyclonal anti-claudin-3 (ab15102), rabbit polyclonal anti-E-cadherin (ab15148), rabbit polyclonal anti-occludin (ab222691); rabbit polyclonal anti-leptin (ab16227, Abcam, Cambridge, UK, dilution 1:200), rabbit monoclonal anti-CD3 (ab16669). The immunoreaction was verified with the negative controls, treated by identical immunohistochemical staining, excluding the application of the primary antibody (Hułas-Stasiak et al, 2020).…”