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A cryo-fixation protocol to study the structure of the synaptonemal complex.
Ortiz, Rosario; Echeverría, Olga M; Masich, Sergej; Höög, Christer; Hernández-Hernández, Abrahan.
Afiliación
  • Ortiz R; Laboratorio de Microscopía Electrónica, Facultad de Ciencias, Universidad Nacional Autónoma de México, Mexico City, Mexico.
  • Echeverría OM; Laboratorio de Microscopía Electrónica, Facultad de Ciencias, Universidad Nacional Autónoma de México, Mexico City, Mexico.
  • Masich S; Department of Cell and Molecular Biology, Biomedicum, Karolinska Institutet, Tomtebodavägen 16, S-171 65, Stockholm, Sweden.
  • Höög C; Department of Cell and Molecular Biology, Biomedicum, Karolinska Institutet, Tomtebodavägen 16, S-171 65, Stockholm, Sweden.
  • Hernández-Hernández A; Biología de Células Individuales (BIOCELIN), Laboratorio de Investigación en Patología Experimental, Hospital Infantil de México Federico Gómez, Mexico City, Mexico. abrahan.h.hernandez@gmail.com.
Chromosome Res ; 30(4): 385-400, 2022 12.
Article en En | MEDLINE | ID: mdl-35486207
ABSTRACT
Genetic variability in sexually reproducing organisms results from an exchange of genetic material between homologous chromosomes. The genetic exchange mechanism is dependent on the synaptonemal complex (SC), a protein structure localized between the homologous chromosomes. The current structural models of the mammalian SC are based on electron microscopy, superresolution, and expansion microscopy studies using chemical fixatives and sample dehydration of gonads, which are methodologies known to produce structural artifacts. To further analyze the structure of the SC, without chemical fixation, we have adapted a cryo-fixation method for electron microscopy where pachytene cells are isolated from mouse testis by FACS, followed by cryo-fixation, cryo-substitution, and electron tomography. In parallel, we performed conventional chemical fixation and electron tomography on mouse seminiferous tubules to compare the SC structure obtained with the two fixation methods. We found several differences in the structure and organization of the SC in cryo-fixed samples when compared to chemically preserved samples. We found the central region of the SC to be wider and the transverse filaments to be more densely packed in the central region of the SC.
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Texto completo: 1 Base de datos: MEDLINE Asunto principal: Complejo Sinaptonémico / Microscopía Idioma: En Revista: Chromosome Res Asunto de la revista: BIOLOGIA MOLECULAR Año: 2022 Tipo del documento: Article

Texto completo: 1 Base de datos: MEDLINE Asunto principal: Complejo Sinaptonémico / Microscopía Idioma: En Revista: Chromosome Res Asunto de la revista: BIOLOGIA MOLECULAR Año: 2022 Tipo del documento: Article