title: Visualizing the mouse podocyte with multiphoton microscopy.
abstract: The podocyte is a highly specialized kidney glomerular epithelial cell that plays an essential role in glomerular filtration and is believed to be the target of numerous glomerular diseases leading to proteinuria. Despite the leaps in our understanding of podocyte biology, new methodologies are needed to facilitate research into the cell. Multiphoton microscopy (MPM) was used to image the nephrin knockout/green fluorescent protein (GFP) knock-in heterozygote (Nphs1(tm1Rkl)/J) mouse. The nephrin promoter restricts GFP expression to the podocytes that fluoresce green under excitation. From the exterior of an intact kidney, MPM can peer into the renal parenchyma and visualize the podocytes that outline the globular shape of the glomeruli. Details as fine as the podocyte's secondary processes can be resolved. In contrast, podocytes exhibit no fluorescence in the wildtype mouse and are invisible to MPM. Phenotypically, there are no significant differences between wildtype and Nphs1(tm1Rkl)/J mice in body weight, urinary albumin excretion, creatinine clearance, or glomerular depth. Interestingly, the glomeruli are closer to the kidney capsule in female mice, making the gender the preferred choice for MPM. For the first time, green fluorescent podocytes in a mouse model free of confounding phenotypes can be visualized unequivocally and in the "positive" by MPM, facilitating intravital studies of the podocyte.
authors.affiliation: Division of Nephrology/Hypertension, Northwestern University, Chicago, IL, USA. khoury.charbel@gmail.com
authors.forename: Charbel C
authors.initials: CC
authors.lastname: Khoury
authors.forename: Mark F
authors.initials: MF
authors.lastname: Khayat
authors.forename: Tet-Kin
authors.initials: TK
authors.lastname: Yeo
authors.forename: Petr E
authors.initials: PE
authors.lastname: Pyagay
authors.forename: Amy
authors.initials: A
authors.lastname: Wang
authors.forename: Allan M
authors.initials: AM
authors.lastname: Asuncion
authors.forename: Kumar
authors.initials: K
authors.lastname: Sharma
authors.forename: Weiming
authors.initials: W
authors.lastname: Yu
authors.forename: Sheldon
authors.initials: S
authors.lastname: Chen
mesh_terms: D000818:Animals; D005260:Female; D055879:Gene Knock-In Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007678:Kidney Glomerulus / Q000166:cytology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D050199:Podocytes / Q000166:cytology* / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; C113267:nephrin; D049452:Green Fluorescent Proteins