Objective
The investigations comprised detailed studies of chromosome synapsis and crossing over in diploids and polyploid species. Aims were to gain further insight into the processes of the genetic regulation of chromosome pairing and crossing over, the extent of chromosome interlocking during synapsis and the short term effects of ionising radiation on chromosomes in the meiotic prophase. The analysis of the biochemical composition of meiosis specific structures has been focussed on the possible role of deoxyribonucleic acid (DNA) topoisomerase II RecA like proteins in meiosis.
THE PROPOSED PROJECT COMPRISES THE FOLLOWING TASKS:
1.-THE ANALYSIS OF THE LONG TERM EFFECT OF RADIATION ON MEIOSIS IN THE HUMAN MALE WILL BE CONTINUED. CONTROL BIOPSIES (75 CASES) PRIOR TO RADIATION TREATMENT HAVE BEEN EMBEDDED FOR ULTRASTRUCTURAL ANALYSIS AND BIOPSIES FROM FOUR MEN HAVING RECEIVED A GONADAL RADIATION DOSE OF 0.4-1.1 GRAY 2.5-3 YEARS AGO DURING TREATMENT FOR CANCER OF THE CONTRALATERAL TESTICLE ARE AT PRESENT UNDER INVESTIGATION. A SUFFICIENT NUMBER OF NUCLEI WILL BE RECONSTRUCTED TO ALLOW A STATISTICAL COMPARISON WITH THE REFERENCE MATERIAL.
2.-THE SHORT TERM EFFECTS OF RADIATION ON MEIOSIS WILL BE ANALYSED IN THE SILKWORM. LARVAE AT DIFFERENT STAGES OF DEVELOPMENT WILL BE SUBJECTED TO RADIATION DOSES SIMILAR TO THOSE USED IN RADIATION THERAPY OF THE HUMAN MALE AND FEMALE AND THE MEIOTIC NUCLEI ANALYSED BY SURFACE SPREADING AND THREE DIMENSIONAL RECONSTRUCTION OF NUCLEI AT DIFFERENT TIME INTERVALS THEREAFTER.
FOR THE FIRST TIME THE RADIATION SENSITIVITY OF SYNAPTONEMAL COMPLEX FORMATION, RESOLUTION OF INTERLOCKINGS AND REDISTRIBUTION OF RECOMBINATION NODULES ENSURING THAT EACH BIVALENT HAS AT LEAST ONE CHIASMA WILL BE EVALUATED.
3.-A MOLECULAR ANALYSIS OF THE COMPOSITION AND ORGANISATION OF THE SYNAPTONEMAL COMPLEX WILL BE ATTEMPTED.BIVALENTS WILL BE ISOLATED FROM THE RECOMBINATION PROFICIENT BOMBYX MALES AND THE RECOMBINATION DEFICIENT FEMALES AND THE CHROMATIM STRIPPED FROM THE SYNAPTONEMAL COMPLEXES BY ENZYMATIC DIGESTION OR MECHANIC DISRUPTION.SUBSEQUENTLY AN ANALYSIS OF THE DNA AND RNA ASSOCIATED WITH THE SYNAPTONEMAL COMPLEX IS PLANNED IN ORDER TO ASSASS THE BIOCHEMICAL BASIS FOR HOMOLOGUE RECOGNITION AND CROSSING OVER.
4.-THE ULTRASTRUCTURAL ANALYSIS OF THE ZYGOTENE STAGE IN THE NORMAL HUMAN FEMALE WILL BE COMPLETED AND A STUDY OF CROSSING OVER AND CHIASMA FORMATION DURING PACHYTENE AND DIPLOTENE UNDERTAKEN. THIS ANALYSIS WILL PROVIDE A DETAILED REFERENCE MATERIAL SIMILAR TO THAT NOW AVAILABLE FOR THE HUMAN MALE.IF IN VITRO CULTIVATED OOCYTES CAN BE MADE AVAILABLE,IT WILL BE RELEVANT TO STUDY THE STAGES UP TO METAPHASE. I.
5.-A COMPARATIVE STUDY OF MEIOSIS BY RECONSTRUCTION OF NUCLEI WILL BE UNDERTAKEN IN THE MALE MOUSE. TO DATE MEIOTIC ANALYSES IN THE MOUSE HAVE BEEN RESTRICTED TO SPREADING OF NUCLEI WITH CHROMOSOMAL ABERRATIONS AND THUS TO STUDIES OF PACHYTENE. THIS IS REGRETTABLE IN VIEW OF THE VERY EXTENSIVE RADIATION PROGRAMME CONDUCTED WITH THIS MAMMAL,ESPECIALLY IN THE UNITED KINGDOM AND THE NETHERLANDS.A DETAILED KNOWLEDGE OF CHROMOSOME PAIRING,FORMATION AND DISTRIBUTION OF RECOMBINATION NODULES AND CHIASMATA WILL HELP TO UNDERSTAND THE SHORT TERM EFFECTS OF RADIATION IN THIS MAMMAL AND SUPPLEMENT THE RESULTS OBTAINED WITH THE SILKWORMS.
6.-THE EFFECT OF CROSSING OVER ON THE STABILISATION OF MULTIVALENT CHROMOSOME CONFIGURATIONS IN POLYPLOID SPECIES WILL BE INVESTIGATED IN TRI- AND TETRAPLOID MALES OF BOMBYX AND IN WHEAT LINES CARRYING MULTIPLE DOSES OF THE PH GENE KNOWN TO CONTROL REGULAR CHROMOSOME DISJUNCTION
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences genetics DNA
- natural sciences biological sciences biochemistry biomolecules proteins
- natural sciences biological sciences zoology mammalogy
- natural sciences biological sciences genetics heredity
- natural sciences biological sciences genetics chromosomes
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Coordinator
2500 KOEBENHAVN
Denmark
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