Complementary freeze fracture replication: An example of its use in the study of horizontal cell gap junctions of the carp retina

✍ Scribed by Tonosaki, Akira ;Washioka, Hiroshi ;Nakamura, Hidenori ;Negishi, Koroku

• Publisher: Wiley (John Wiley & Sons)
• Year: 1985
• Tongue: English
• Weight: 635 KB
• Volume: 2
• Category: Article
• ISSN: 0741-0581
• DOI: 10.1002/jemt.1060020303

✦ Synopsis

Dark-adapted carp retinas were fixed with glutaraldehyde, frozen in liquid nitrogen slush, and their complementary replicas were harvested using a bipolar-vacuum evaporater. On montage pictures, plasmic (PFF) and ectoplasmic fracture faces (EFF) of the external horizontal cells (EHC) were marked, and PFF of gap junction (GJ) areas were scrutinized by virtue of particulate aggregation visible at relatively low magnification. Summation of digitized values obtained from a complementary pair of replicas was taken to represent all existent G J areas in a n examined EHC surface. A similar method was applied to large horizontal processes (LHP) that are located in the innermost part of the internal nuclear layer and currently accepted to be extensions of EHC. The occurrence of fractured faces of plasma membrane was smaller in EHC than LHP. The percentage of GJ areas was about ten times larger in EHC than LHP. Examining complementary replicas at relatively low magnification appears to be useful for facilitating quantitative analysis of intercellular coupling by G J structures.