Feedback-controlled constant-pressure anterior chamber perfusion in live mice

摘要

Purpose: To describe live mouse, anterior chamber constant-pressure perfusion by an approach using feedback-controlled coupling of pressure and flow to maintain a preset pressure.Methods: We established a microperfusion system that maintains a constant preset pressure in the anterior chamber of live mice by automatically regulating the microsyringe pump flow rate with a computer-controlled voltage feedback loop. Perfusion was by single-needle cannulation. We characterized the following in C57BL/6 mice aged 3–4 months in vivo: (i) pressure stability, (ii) pressure and flow rate reproducibility, (iii) total outflow facility, and (iv) anterior segment histology after perfusion.Results: Twenty live mice underwent perfusion. Constant pressure was quickly attained and stably maintained. The coefficient of pressure variation over time during perfusion at a preset pressure was <0.001. The average coefficient of variation for repeat pressure and flow rate measurements was 0.0005 and 0.127, respectively. The relationship between flow rate and pressure was linear for perfusions between 15 and 35 mmHg. The total outflow facility was 0.0066 µl/min/mmHg. Perfusion system resistance (0.5 mmHg/min/µl) was negligible relative to the ocular outflow resistance (147 mmHg/min/µl) at physiologically relevant perfusion pressures of 15–35 mmHg. No histological disruption of the drainage tissue was seen following perfusion.Conclusions: Predetermined pressure was stably maintained during constant-pressure perfusion of live mouse eyes by a method using feedback-controlled coupling of pressure and flow along with single-needle anterior chamber cannulation. Perfusion measurements were reproducible. This approach is potentially useful for exploring aqueous drainage tissue biology, physiology, and pharmacology in live mice.