SUB-PROJECT 3 WILL AIM TO ANALYSE THE EFFECTS OF BISPHENOL A, EE2 AND CIMETIDINE ON MALE GERMLINE CELLS, USING ZEBRAFISH AS A MODEL, SINCE IT ALLOWS UNVIABLE EXPOSURE PROTOCOLS IN COMMERCIAL SPECIES. PATERNAL CONTRIBUTION IS CRUCIAL TO THE CONTROL OF EARLY EMBRYONIC DEVELOPMENT THROUGH THE EXPRESSION OF CERTAIN GENES, THE TRANSCRIPTION OF CERTAIN MSRNAS AND THE MAINTENANCE OF A CORRECT EPIGENETIC PATTERN, WHICH IS REPRODUCED IN THE EMBRYO 24HPF. EMERGING POLLUTANTS CAN MODIFY THIS INFORMATION AND AFFECT, AS HAS ALREADY BEEN SHOWN, THE DEVELOPMENT OF F1. OUR STUDY WILL ANALYSE POTENTIAL DAMAGE TO THE GENOME, TRANSCRIPTOME, AND EPIGENOME IN ANIMALS EXPOSED TO TOXICS DURING EMBRYONIC OR ADULT LIFE, IDENTIFYING GENES FOR GENOTOXIC AND EPIGENOTOXIC DAMAGE. FOR THIS PURPOSE TECHNIQUES WILL BE USED DEVELOPED IN OUR LABORATORY, COLLABORATING WITH OTHER TEAMS FOR THE REALISATION OF THE RRBS. THESE STUDIES WILL ALLOW US TO IDENTIFY THE MOST SENSITIVE PHASES AND THE ELEMENTS POTENTIALLY AFFECTED BY TOXIC AGENTS, IDENTIFYING THE MOST VULNERABLE GENES OR PROMOTERS AND ESTABLISHING A PANEL OF DAMAGE INDICATORS IN THE GERM LINE. _x000D_ in the DESCENDENCE OF EXPUEST PROGENITORS will be evaluated morphogenesis, with special attention to HEART FORMATION, reproductive capacity, IMMUNE RESPERTY, TEJID REGENERATION CAPACITY AND Aging Tax, using MORFOLOGICAL, FUNCIONAL and GENIC EXPRESSION STUDIES. THE POSSIBLE TRANSMISSION OF PATERNAL INFORMATION TO THE CELLS OF THE GERMINAL LINE OF THE F1, WHICH PROVOCARIAN A TRANSGENERATIONAL INHERITANCE TO F2 WAS ANALYSED BY ANALYSING THE GERMLINE CELLS OF THE PREVIOUSLY DEFINED TOXICITY MARKERS. IF APPLICABLE, THE DEVELOPMENT OF F2 WILL ALSO BE EVALUATED USING THE SAME PARAMETERS AS WITH F1. THE POSSIBLE NUTRITIONAL RESCUE OF EPIGENOTOXIC DAMAGE, WHICH MAY BE DUE TO LOWER METHYL TRANSFERASE ACTIVITY OR REDUCED AVAILABILITY OF METHYL GROUPS, WILL ALSO BE EVALUATED THROUGH THE ADMINISTRATION OF FOLATE-RICH DIETS. The INFLUENCE of the somatic microenvironment IN THE RECOVERY OF TOXIC EFFECTS ON THE GERMINAL LINE CELLS IS STUDIATED FOR THE TRANSPLANT OF GERMINAL CELULS EXPUESED AND ANIMAL CONTROL PREVIENTLY STERILISED by ELIMINATION OF YOUR GERMINAL CELLS WITH morfolin DND._x000D_ for the realisation of the state, WILD TYPE (AB) and TRANSGENICAS (VASA:EGFP) will be used, which will allow the isolation, transPLANT AND STUDY OF THE EVOLUTION OF THE GERMINAL CELLS