Domestication Effects on Stress Induced Steroid Secretion and Adrenal Gene Expression in Chickens

Understanding the genetic basis of phenotypic diversity is a challenge in contemporary biology. Domestication provides a model for unravelling aspects of the genetic basis of stress sensitivity. The ancestral Red Junglefowl (RJF) exhibits greater fear-related behaviour and a more pronounced HPA-axis reactivity than its domesticated counterpart, the White Leghorn (WL). By comparing hormones (plasmatic) and adrenal global gene transcription profiles between WL and RJF in response to an acute stress event, we investigated the molecular basis for the altered physiological stress responsiveness in domesticated chickens. Basal levels of pregnenolone and dehydroepiandrosterone as well as corticosterone response were lower in WL. Microarray analysis of gene expression in adrenal glands showed a significant breed effect in a large number of transcripts with over-representation of genes in the channel activity pathway. The expression of the best-known steroidogenesis genes were similar across the breeds used. Transcription levels of acute stress response genes such as StAR, CH25 and POMC were upregulated in response to acute stress. Dampened HPA reactivity in domesticated chickens was associated with changes in the expression of several genes that presents potentially minor regulatory effects rather than by means of change in expression of critical steroidogenic genes in the adrenal.Funding agencies: Swedish Research Council (VR) [621-2011-4731]; Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (FORMAS) [221-2011-1088]; SRC [621-2011-5523]; ERC [322206]; Swedish Centre of Excellence in Animal Welfare</p

A fast ultra performance supercritical fluid chromatography-tandem mass spectrometric method for profiling of targeted phytosterols

Phytosterols are essential structural components of plant cell membranes and possess health-related benefits, including lowering blood cholesterol levels in humans. Numerous analytical methods are being used to profile plant and animal sterols. Chromatography hyphenated to tandem mass spectrometry, is a better option due to its specificity, selectivity, and sensitivity. An ultra-performance supercritical fluid chromatography hyphenated with atmospheric pressure chemical ionization (APCI) tandem mass spectrometric method was developed and eval-uated for fingerprint analysis of seven phytosterols. Mass spectrometry fragmentation behavior was used for phytosterol identification, and multiple reaction monitoring scanning was utilized for phytosterol confirmation, where APCI outperformed superiority in terms of ion intensity, particularly in the production of [M + H-H2O]+ ions rather than [M + H]+ ions. The chromatographic conditions were thoroughly evaluated, and the ionization parameters were optimized as well. In a 3 min. run, the seven phytosterols were separated concurrently. The calibration and repeatability tests were conducted to check the instrument's performance, and the results indicated that all of the phytosterols tested had correlation coefficients (r2) greater than 0.9911 over the con-centration range of 5-5000 ng/mL. The limit of quantification was below 20 ng/mL for all the tested analytes except for stigmasterol and campesterol. The partially validated method was applied for the evaluation of phytosterols in pure coconut oil and palm oil in order to demonstrate its applicability. Total sterols in coconut and palm oils were 126.77 ng/mL and 101.73 ng/mL, respectively. In comparison to earlier methods of phytosterol analysis, the novel method offers a far faster, more sensitive, and more selective analytical process