Chromatin condensation during Scrobicularia plana spermiogenesis: a controlled and comparative enzymatic ultracytochemical study

In Scrobicularia plana testis, a nuclear acid phosphatase (ACPase) activity was detected in mid and late spermatids with the improved Gomori-chloride procedure, Lead deposits were first observed in mid spermatids at focal poi nts over condensed chromatin strands, increasing in density as chromatin fu rther condensated, In late spermiogenesis, lead deposits became concentrate d between chromatin aggregates, and after total DNA compaction were transfe red to the nuclear periphery and then shed into the cytoplasm. The specific ity of the nuclear ACPase was tested against different pH values (3.9, 7.2, 7.8, 9.0), substrates (TPP, IDP, TMP, p-NCS, ATP, GTP, AMP, ADP, AMP-PNP) and inhibitors (NaF, levamisole, Zn, vanadate, theophylline). To further sp ecify the nature of this nuclear ACPase, other enzymes were comparatively s tudied at their optimal pH values and at pH 5.0. nucleoside-diphosphatase, thiamin-pyrophosphatase, inorganic trimetaphosphatase, lysosomal arylsulfat ases A and B, ATPase, GTPase, 5'-nucleotidase, adenylate kinase, and adenyl ate cyclase, Several other controls were introduced to exclude artefactual deposits induced by lead ions and tissue molecules. The results showed that the enzyme has an optimal pH at 5.0, a high specific affinity for beta-GP, and is inhibited by NaF, which suggests that it behaves as a type B-ACPase , and all controls demonstrated the specificity of the enzymic activity. Be cause lead deposits were specifically and temporally associated with sperma tid chromatin condensation, when DNA and RNA synthesis, histones, phosphopr oteins and RNA molecules strongly decrease, it is possible to suggest that the nuclear ACPase could be associated with DNA processing during chromatin compaction or involved in the hydrolysis of 2' and 3' nucleotides resultin g from nuclear RNase action during RNA degradation. (C) 2000 Harcourt Publi shers Ltd.