Transplantation and Re-transplantation of Mouse Kidney Grafts to Study Local Immune Responses

Mouse kidney transplantation is widely used to study the immune response to allogeneic grafts. This response includes a circulating systemic compartment and a resident non circulating one. A distinction between these compartments remains an important caveat to the interpretation of the resident or local immune response’s importance and function. Here, we describe re-transplantation as a method to functionally test the resident component of the primary immune response while also studying the secondary recipient’s response. Our detailed, stepwise protocol can be reliably replicated for both the primary and secondary, donor and recipient operations. The techniques in this protocol can be efficiently implemented by an individual proficient in mouse kidney transplantation surgical procedures.

The Hydrogen-Coupled Oligopeptide Membrane Cotransporter Pept2 is SUMOylated in Kidney Distal Convoluted Tubule Cells

Protein post-translational modification by the Small Ubiquitin-like MOdifier (SUMO) on lysine residues is a reversible process highly important for transcription and protein stability. In the kidney, SUMOylation appears to be important for the cellular response to aldosterone. Therefore, in this study, we generated a SUMOylation profile of the aldosterone-sensitive kidney distal convoluted tubule (DCT) as a basis for understanding SUMOylation events in this cell type. Using mass spectrometry-based proteomics, 1037 SUMO1 and 552 SUMO2 sites, corresponding to 546 SUMO1 and 356 SUMO2 proteins, were identified from a modified mouse kidney DCT cell line (mpkDCT). SUMOylation of the renal hydrogen-coupled oligopeptide and drug co-transporter (Pept2) at one site (K139) was found to be highly regulated by aldosterone. Using immunolabelling of mouse kidney sections Pept2 was localized to DCT cells in vivo. Aldosterone stimulation of mpkDCT cell lines expressing wild-type Pept2 or mutant K139R-Pept2, post-transcriptionally increased Pept2 expression up to four-fold. Aldosterone decreased wild-type Pept2 abundance in the apical membrane domain of mpkDCT cells, but this response was absent in K139R-Pept2 expressing cells. In summary, we have generated a SUMOylation landscape of the mouse DCT and determined that SUMOylation plays an important role in the physiological regulation of Pept2 trafficking by aldosterone.

Transplantation and Re-transplantation of Mouse Kidney Grafts to Study Local Immune Responses

Mouse kidney transplantation is widely used to study the immune response to allogeneic grafts. This response includes a circulating systemic compartment and a resident non circulating one. A distinction between these compartments remains an important caveat to the interpretation of the resident or local immune response’s importance and function. Here, we describe re-transplantation as a method to functionally test the resident component of the primary immune response while also studying the secondary recipient’s response. Our detailed, stepwise protocol can be reliably replicated for both the primary and secondary, donor and recipient operations. The techniques in this protocol can be efficiently implemented by an individual proficient in mouse kidney transplantation surgical procedures.

The Value of CTA Based on Gold Nanorod Contrast Agent in Coronary Artery Diagnosis and Plaque Property Analysis

Coronary CT angiography (CTA) with the characteristics of noninvasive and simple operation is widely used in the diagnosis of coronary artery stenosis. The choice of contrast agent exerts an important impact on the imaging quality of CTA. Conventional iodine contrast agents are easily excreted by the kidneys, from which the imaging window is short, and the imaging quality is poor. Metal nanomaterials have unique optical properties and have broad application prospects in imaging. Our aim is to explore the value of gold nanorod contrast agent in the diagnosis of coronary heart disease. A gold nanorod suspension was first prepared, and the prepared gold nanorod was uniform and had good dispersibility. It can be seen from the light absorption curve that there are two obvious peaks on the UV absorption peak of the gold nanorods. The gold nanorods were cultured in different solutions, and it was found that the particle size of the gold nanorods did not change significantly within 72 hours, indicating that the prepared gold nanorods had good stability. When observing the damage degree of mouse kidney tissue, it was shown that the damage degree of gold nanorod contrast agent to mouse kidney tissue was less than that of iodine contrast agent. The above results indicate that the gold nanorod contrast agent has good stability and safety. Therefore, our study demonstrated that the gold nanorod contrast agent has high value in the diagnosis of coronary arteries and the analysis of plaque properties.