Microtubule organization and the distribution of gamma-tubulin in spermatogenesis of a beetle, Tenebrio molitor (Tenebrionidae, Coleoptera, Insecta)

Abstract

The present study focuses on the restructuring of the microtubule (MT) cytoskeleton and microtubule-organizing centres (MTOCs) throughout spermatogenesis of a darkling beetle, Tenebrio molitor (Tenebrionidae, Coleoptera, Insecta). To this end, serial ultrathin sections through male germ cells were studied using transmission electron microscopy. Additionally, spindles and young spermatids were isolated from testes under MT-stabilizing conditions and doubly labeled with antibodies against beta- and gamma-tubulin. The latter is a tubulin isoform detected in MTOCs of a wide variety of species. The observations suggest that microtubules may be nucleated from sites with and without high gamma-tubulin content and that these sites do not necessarily possess canonical centrosomes. In a prominent cytoplasmic MT system of primary spermatocytes in prophase, microtubule nucleation apparently occurs in the absence of immunologically detectable gamma-tubulin. At the poles of meiotic spindles, MTs are directly inserted into gamma-tubulin-containing material and this connection is considered responsible for their nucleation. The interzone spindle MTs of telophase cells contain gamma-tubulin and this may confer stability to them. Finally, manchette MTs of spermatids originate in the vicinity of the acrosome precursor but are not inserted into this body. The acrosome precursor is surrounded by a membrane and is clearly detected by the antibody against gamma-tubulin.