Localization of SCP2 and SCP3 protein molecules within synaptonemal complexes of the rat.

Abstract

SCP2 and SCP3 are major protein components of the lateral elements (LEs) of synaptonemal complexes (SCs) of the rat, with Mrs of 173, 000 and 30,000. We performed a detailed immunocytochemical comparison of the localization of SCP2 and SCP3 within SCs at the electron microscopic level. The ultrastructural localization of SCP2 and SCP3 was analyzed by immunogold labeling of two types of preparations, namely surface-spread spermatocytes and ultrathin sections of Lowicryl-embedded testicular tissue of the rat. For each of the antisera used, the distribution of immunogold label over SCs in surface-spread spermatocytes differed significantly from the distribution of label on sections. We attributed this difference to artifacts caused by the surface-spreading technique, and therefore we relied on sections for the precise localization of epitopes. On sections, the distribution of label obtained with two antisera against nonoverlapping, widely separated fragments of SCP2 did not differ significantly. There was a small but significant difference between the labeling pattern obtained with an anti-SCP3 serum and the pattern obtained with either of the two antisera against fragments of SCP2; although for all three antisera the peak of the immunogold label coincided with the center of the LE, the distributions of label obtained with the antisera against fragments of SCP2 were asymmetrical, with a shoulder at the inner side of the LE, whereas the distribution of label obtained with anti-SCP3 serum was symmetrical. Furthermore, we observed fuzzy connections between the LEs that were labeled by anti-SCP2 but not anti-SCP3 antibodies. It is possible that labeling of these fuzzy bridges caused the shoulder in the gold label distributions obtained with anti-SCP2 antibodies.