FIXATION OF THE MACULA DENSA WITH FIXATIVES OF DIFFERENT OSMOLARITIES IN NORMAL AND DIABETIC RATS

The influence of different fixative osmolarities on the ultrastructure of the macula densa region has been studied in normal and in streptozotocin diabetic rats after 50 days' duration of diabetes. In vivo perfusion fixation of the kidneys was performed retrogradely through the aorta in five groups of controls and diabetic animals with a 1% glutaraldehyde fixative in a Tyrode solution. Between groups, the osmolarity of the fixative solution varied from 216 to 476 mosm/1. This was accomplished by variation of the NaCl concentration of the fixative vehicle. The tissue was embedded in Epon and midcortical tissue was prepared for electron microscopy. The volume density of the lateral intercellular spaces between the macula densa cells was measured applying a morphometric technique. The results confirmed that the lateral intercellular spaces are narrowed in diabetes and also showed that for all fixative osmolarities the difference is persistent between diabetic and non-diabetic animals, except at very low osmolarities. The abnormal macula densa ultrastructure has been suggested to be connected with the functional abnormalities in diabetes, i.e. the resetting of the tubuloglomerular feedback and subsequent increases in GFR. It is therefore of interest to further understand the mechanisms underlying the narrowing of the lateral intercellular spaces in diabetes. The present study has shown that the size difference in lateral intercellular spaces between diabetic and normal animals is reproducible over a wide range of fixative osmolarities, further indicating that it is an in vivo phenomenon. The diminution of the LIS in diabetes could perhaps indicate a decrease in signal function, previously reported at the MD level in diabetic animals, contributing to the increased GFR in diabetes.