Project description:Abstract Objective: This study was designed to reveal the potential molecular mechanisms of long-term overgrazing-induced dwarf of sheepgrass (Leymus chinensis). Methods: The electrospray ionization-mass spectrometry (ESI-MS) system was used to generate the proteomic data in dwarf sheepgrass from long-term overgrazed rangeland and normal sheepgrass from long-term enclosed rangeland. The differentially expressed proteins (DEPs) between dwarf and normal sheepgrass were identified, following the analyses of potential functions and interactions of DEPs. Additionally, the expressions of DEPs were confirmed by high performance liquid chromatography-mass spectrum (HPLC-MS) system with multiple reaction monitoring (MRM) method. Results: A total of 51 up-regulated proteins and 53 down-regulated proteins were identified in the dwarf sheepgrass, compared with the normal controls. Some proteins like SAT5_ARATH and DAPA_MAIZE were enriched in the pathway of amino acid biosynthesis. In the protein-protein interaction network, RPOB2_LEPTE, A0A023H9M8_9STRA, ATPB_DIOEL, RBL_AMOTI and DNAK_GRATL interacted with each other. Furthermore, 4 modules were extracted from the PPI network. In addition, the HPLC-MS analysis confirmed the up-regulation of ATPB_DIOEL and the down-regulation of DNAK_GRATL in dwarf samples compared with the controls. Conclusion: The up-regulated ATPB_DIOEL and down-regulated DNAK_GRATL, as well as the proteins that interacted with them, such as RPOB2_LEPTE, A0A023H9M8_9STRA and RBL_AMOTI, may be associated with the long-term overgrazing-induced sheepgrass dwarf.
Project description:Kinase activity of cGMP-dependant, type II, protein kinase (PRKG2) is required for the proliferative to hypertrophic growth transition of growth plate chondrocytes during endochondral ossification. Loss of PRKG2 function in rodent and bovine models results in dwarfism. We compared growth plate cartilage gene expression profiles of PRKG2R678X/R678X, dwarf and PRKG2R678X/+, unaffected Angus cattle using microarray technology to discover pathways regulated by PRKG2. Calves used in this analysis were produced by the mating between a dwarf carrier sire and two dams- a mother (carrier) and daughter (dwarf) pair. Each dam produced three calves by embryo transfer, which were born in two calving groups. Growth plate cartilage was harvested from the tibia of each animal at approximately 215 days of age. Total RNA was extracted by a modified Trizol protocol. The bovine cDNA microarray (GEO Accession: GPL8813) was used for this analysis. Samples were labeled with Cy3 or Cy5 and hybridized by dye swapping the dwarf (4 dwarf animals- 2 per dam/calving group) and unaffected individuals (2 unaffected animals- 1 per dam/ calving group) such that each dwarf was compared to the unaffected full-sib produced in the same calving group. Each cDNA array was analyzed at three different PMT levels (PMT70,80 and 90), resulting in 12 total image files for statistical analysis. Dwarf PRKG2 R678X homozygous cattle were compared to unaffected, age matched samples. Samples were labeled with Cy3 or Cy5 and hybridized by dye swapping the dwarf (4 dwarf animals- 2 per dam/calving group) and unaffected individuals (2 unaffected animals- 1 per dam/ calving group) such that each dwarf was compared to the unaffected full-sib produced in the same calving group (i.e. unaffected samples had 2 technical replicates). Each cDNA array was analyzed at three different PMT levels (PMT70,80 and 90), resulting in 12 total image files for statistical analysis.