title: Transmission electron microscopy, immunocytochemical and fluorescence in situ hybridisation studies in a case of 100% necrozoospermia: case report.
abstract: We present the ultrastructural, functional and chromosomal analyses of sperm from an infertile man with a normal karyotype and 100% necrozoospermia. Tests for microbial infection showed the presence of Escherichia coli in seminal and urethral fluid. Semen analysis was performed concomitantly with the infection and repeated twice after the therapy, in the absence of infection, to control the possible recovery of necrozoospermia. Morphological sperm evaluation was performed by light, fluorescent and electron microscopy; meiotic segregation was examined by fluorescence in situ hybridisation (FISH) technique. Polymerase chain reaction (PCR) was carried out on DNA from peripheral blood lymphocytes to analyse partial sequences of the Akap4 and Akap3 genes. AKAP3 protein is synthesised in round spermatids, incorporated into the fibrous sheath and is involved in organising the basic structure of the fibrous sheath. AKAP4 protein is incorporated late in spermatid development and plays a major role in completing fibrous sheath assembly. The whole sperm population was immotile and non-viable. The ultrastructural characteristics of sperm necrosis were identified. Immunostaining of tubulin and AKAP4 proteins was negative in sperm tails. PCR did not reveal any deletions in the investigated regions. FISH sperm analysis highlighted an altered meiotic segregation. After recovery from infection, 100% necrozoospermia persisted and chromosomal sperm aneuploidies were still present.
authors.affiliation: Department of Surgery, Biology Section, University of Siena, Siena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Moretti
authors.forename: B
authors.initials: B
authors.lastname: Baccetti
authors.forename: G
authors.initials: G
authors.lastname: Scapigliati
authors.forename: G
authors.initials: G
authors.lastname: Collodel
mesh_terms: D054758:A Kinase Anchor Proteins; D004247:DNA / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D007248:Infertility, Male / Q000503:physiopathology; D008297:Male; D008540:Meiosis; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D009336:Necrosis / Q000235:genetics / Q000473:pathology; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D013076:Sperm Count; D013094:Spermatozoa / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure*; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
chemical_list: D054758:A Kinase Anchor Proteins; C485480:AKAP4 protein, human; D011498:Protein Precursors; D014404:Tubulin; D004247:DNA