Quantitative proteomic analysis of aqueous humor after rabbit lensectomy reveals differences in coagulation and immunomodulatory proteins

Abstract

Proteomics of juvenile rabbit aqueous humor was evaluated before and after surgery to determine proteins responsible for postoperative outcomes.

Tissue Plasminogen Activator Effects on Fibrin Volume and the Ocular Proteome in a Juvenile Rabbit Model of Lensectomy

“…A total of 27 New Zealand White rabbits (Kuiper Rabbit Ranch, Gary, IN), 6 to 7 weeks of age and weighing between 600 and 900 g, were housed in a 14-hour light/10-hour dark cycle and fed a diet of pelleted chow (Laboratory Rabbit Diet High Fiber 5326; LabDiet, St. Louis, MO) daily and a Timothy hay cube twice per week (Bio-Serv, Flemington, NJ). Based on previous studies with preventative pharmacologic therapies where five or six rabbits per group showed significant postoperative differences clinically and in AH proteins, 14 , 15 eyes of either male or female sex per group were chosen to be randomized to the tPA or control balanced salt solution (BSS) group for postoperative treatments (described below). Rabbits were anesthetized with isoflurane (5% induction, 2.5%–4% maintenance) with 2 L/min of oxygen for surgery and postoperative exams.…”

“…POD3 was chosen because (1) we previously classified this day as the peak of the postoperative response, and (2) it allowed us to compare outcomes with our previous clinical and proteomic studies. 14 , 15 The intraocular pressure (IOP) in each eye was measured using a TonoLab tonometer (Icare USA, Raleigh, NC). Slit-lamp biomicroscopy (SL-D8Z Digital-Ready Zoom Slit Lamp; Topcon Medical Systems, Oakland, NJ) was performed with photographs obtained using a Nikon D810 camera (Nikon USA, Melville, NY) to evaluate each eye in the categories of iris synechiae, pupil dilation in millimeters, and presence or absence of hemorrhage in the anterior chamber.…”

“…Our laboratory utilizes a juvenile rabbit animal model to investigate the response following intraocular surgery. 14 , 15 Juvenile rabbits have a robust immune and fibrotic response following lensectomy with IOL insertion, making them an ideal animal for studying treatment outcomes following intraocular surgery. Sampling of the aqueous humor (AH) before and after a surgical intervention can help deduce the mechanisms and proteins responsible for the exaggerated immune response.…”

“…Using mass spectrometry (MS), we have previously shown that several coagulation factors and inflammatory proteins are significantly increased in abundance in the AH following lensectomy using our rabbit animal model. 14 In this current study, we determine changes in protein abundance of 33 proteins 24 hours following tPA administration. By understanding these abundance changes, we gain insights regarding the effect of tPA on the ocular proteome and which pathways may contribute to postoperative complications.…”

“…Determining how the ocular proteome responds to pharmacotherapies builds upon our previous work establishing the ocular proteome in an untreated rabbit model. 14 …”