Antifreeze Glycoproteins from Antarctic Notothenioid Fishes Fail to Protect the Rat Cardiac Explant during Hypothermic and Freezing Preservation

Abstract

The antarctic notothenioid fishes avoid freezing through the action of circulating antifreeze glycoproteins (AFGPs). This study investigated whether AFGPs could serve as cryoprotectants for the isolated rat heart under three different storage conditions. (1) Hearts were flushed with 15 mg AFGP/ ml cardioplegic solution (CP) and stored for 9 h at 0°C. This AFGP concentration has been reported to protect pig oocytes during hypothermic storage. (2) Hearts were flushed with 10 mg AFGP/ml CP-14 and stored frozen at - 1.4°C for 3 h. At this concentration the AFGPs significantly reduce the solution freezing point and also change the crystal morphology from dendritic to spicular. (3) Hearts were flushed with 10 μg AFGP/ml CP-15 and stored frozen at - 1.4°C for 5 h. At this low concentration the AFGPs have a strong inhibitory effect on ice recrystallization, but have little effect on the freezing point and less apparent effect on the crystal habit. After hypothermic or freezing storage, the functional viability was assessed by determining cardiac output (CO) during working reperfusion. No difference in CO was found between AFGP-treated and untreated hearts after 9 h of 0°C storage. CO in hearts frozen in CP-14 without AFGPs recovered to 50% of the freshly perfused control hearts. Hearts frozen in the presence of high concentrations of AFGPs (10 mg/ml CP-14) failed to beat upon thawing and reperfusion, although their tissue ice content was less than that found in hearts without AFGP treatment. Hearts frozen with low concentrations of AFGPs (10 μg/ml CP-15) showed reduced recovery upon thawing and reperfusion compared to CP-15 hearts, which recovered to 67% of freshly perfused controls. Thus notothenioid fish AFGPs not only fail to enhance storage of the isolated rat heart preparation at hypothermic temperatures, but cause increased damage under freezing conditions regardless of AFGP concentration.