The Research Of Male Zebrafish Holding Water Attracts Females And Induces Spawning Behavior

Reproduction is a continuation of the biological gene race,and is also a vital life activity.Evolution has given rise to the formation of a variety of reproductive mechanisms.In animals,spermatozoa fertilize an egg by combining their genes with those of the egg,a very common strategy.In this process,a series of complex reproductive mechanisms,consisting of the generation,release,and combination of spermatozoa and eggs,are influenced by various environmental factors,such as day-length and temperature.However,biological factors,such communication between the sexes,are also critical.In this study,in order to isolate and identify important factors that affect zebrafish reproduction-sex pheromones-we have used behavioral,chromatographic mass spectrometry(GC-MS),high performance liquid chromatography(HPLC)and olfactory electro-physiologal(EOG)approaches.Firstly,we assessed the relative importance of visual and olfactory stimuli on zebrafish reproductive behavior,namely pre-spawning attraction and spawning behavior of females.Male zebrafish holding water can attract females and can evoke them to spawn.Before the females spawn,we found that they were attracted to male-released odorants.Sexually mature male and female wild-type zebrafish were used as donors and receivers,respectively,for visual and stimuli.Female zebrafish were attracted to males that they could smell but not see more than to males that they could see but not smell.However,the effects of the two stimuli were additive.In the spawning behavior experiment,we also used in the sexual maturity of wild-type male and female zebrafish for visual and male zebrafish two different stimuli,with non-visual non-exchange of holding water in the natural state of the female sexual attraction percentage.As a blank control,by comparing the visual and male zebrafish holding water with two different stimuli,as well as female zebrafish in the visual and male zebrafish double stimulus,the number of spawning females was determined in the act of causing female zebrafish to spawns.The stimulation of male zebrafish holding water dominates the visual stimulus.In the second part,we carried out GC-MS analysis of the co-existence of male zebrafish and female zebrafish.System Based on the study of the zebrafish reproductive system neutral pheromone,we boldly assumed that 36 steroids were analyzed.According to the results of GC-MS analysis,there are 22 kinds of steroids in the holding water when male zebrafish and female zebrafish coexist,and there are 10 kinds of steroids that may exist.In order to verify whether these steroids are substances that attract female zebrafish,we use these substances to stimulate female zebrafish and record the olfactory electrophysiological response(EOG)of female zebrafish.As a result,none of the 22 steroids had a continuous olfactory electrophysiological response,and we were not sure of attracting female zebrafish and promoting the production of female zebrafish.In the third part,we carried out high performance liquid chromatography(HPLC)analysis of holding water,male zebrafish holding water and female zebrafish fed with common zebrafish and female zebrafish.Based on a comparative analysis of three high performance liquid chromatograms,we found that the holding water of male zebrafish and female zebrafish contained all of the substances in the water when they were present alone.But the peak area is not male and female zebrafish peak map area simply added,indicating that female zebrafish and male zebrafish co-exist,mutual promotion of each other's secretion of certain substances.In order to determine the attracting of female zebrafish and the spawning of substances,we used the zebrafish three groups of holding water samples(32 isolates per group)of the 96 components of the female zebrafish olfactory electrophysiology(EOG)experiments.According to the electrophysiological results,it was found that the components of female zebrafish olfactory electrophysiological reaction existed at 10 min and 20 min,and the female zebrafish had a strong olfactory electrophysiological reaction when the male zebrafish and female zebrafish existed,And more strongly than male zebrafish female zebrafish alone,once again proved that they coexist each other to promote the secretion of certain substances.And the female zebrafish on the male zebrafish holding water separation component olfactory electrophysiological reaction was significantly stronger than the female holding water separation components to be strong,indicating that attracting female and promote the female zebrafish spawning material exists in the male zebrafish breeding Water.Comprehensive experiments show that male zebrafish holding water can attract female zebrafish,and can cause female zebrafish to spawning.And when the female zebrafish is present,it can promote the secretion of male zebrafish to attract female zebrafish and cause spawning.These substances may be present in the holding spectrum of male zebrafish for 10 min and 20 min of substances or mixtures,which is our next research direction.