The role of extracellular vesicles in animal reproduction and diseases

Extracellular vesicles(EVs)are nanosized membrane-enclosed compartments that serve as messengers in cell-to-cell communication,both in normal physiology and in pathological conditions.EVs can transfer functional proteins and genetic information to alter the phenotype and function of recipient cells,which undergo different changes that positively affect their structural and functional integrity.Biological fluids are enriched with several subpopulations of EVs,including exosomes,microvesicles(MVs),and apoptotic bodies carrying several cargoes,such as lipids,proteins,and nucleic acids.EVs associated with the reproductive system are actively involved in the regulation of different physiological events,including gamete maturation,fertilization,and embryo and fetal development.EVs can influence follicle development,oocyte maturation,embryo production,and endometrial-conceptus communication.EVs loaded with cargoes are used to diagnose various diseases,including pregnancy disorders;however,these are dependent on the type of cell of origin and pathological characteristics.EV-derived microRNAs(miRNAs)and proteins in the placenta regulate inflammatory responses and trophoblast invasion through intercellular delivery in the placental microenvironment.This review presents evidence regarding the types of extracellular vesicles,and general aspects of isolation,purification,and characterization of EVs,particularly from various types of embryos.Further,we discuss EVs as mediators and messengers in reproductive biology,the effects of EVs on placentation and pregnancy disorders,the role of EVs in animal reproduction,in the male reproductive system,and mother and embryo cross-communication.In addition,we emphasize the role of microRNAs in embryo implantation and the role of EVs in reproductive and therapeutic medicine.Finally,we discuss the future perspectives of EVs in reproductive biology.

Interstitial tissue-specific gene expression in mouse testis by intra-tunica albuguineal injection of recombinant baculovirus

The purpose of this study is to establish a gene delivery system for interstitial tissue-specific protein expression in mice testes using modified recombinant baculovirus. Green fluorescent protein （GFP）-expressing recombinant baculovirus （GFP-baculovirus）, in which the insect cell-specific polyhedron promoter was replaced by the cytomegalovirus （CMV）-IE promoter, was used to transfect testicular cells in vitro, and for intra-tunica albuguineal injection of the interstitial tissue of the testis. GFP expression was monitored in frozen testes sections by fluorescence microscopy. Expression of GFP in testicular tissues was also assessed by reverse transcription polymerase chain reaction （RT-PCR）, and protein expression was assessed by Western blot. Testicular cells in vitro were infected efficiently by modified recombinant GFP-baculovirus. lntra-tunica albuguineal injection of GFP- baculovirus into the mouse testis resulted in a high level of GFP expression in the interstitial tissues. RT-PCR analysis clearly showed GFP gene expression in the testis, particularly interstitial tissues. Intra-tunica albuguineal injection of a modified baculovirus that encoded recombinant rat insulin-like growth factor binding protein （IGFBP）-5 resulted in an increase in IGFBP-5 in testis and semen. In conclusion, we have developed an efficient delivery system for gene expression in vivo in testicular cells, particularly cells of the interstitial tissue using intratunica albuguineal injection of a modified recombinant baculovirus. This method will be particularly relevant for application that requires gene delivery and protein expression in the testicular cells of the outer seminiferous tubule of the testis.