栉孔扇贝LAMP1基因的序列特征、表达模式及功能OA

Lysosome-associated membrane protein 1(LAMP1)is a member of the LAMP family;current studies on this gene have mainly focused on vertebrates,wherein it is considered to be involved in spermatogenesis.To date,no studies have examined LAMP1 in mollusks.Based on preliminary omics analyses conducted in our laboratory,LAMP1 is presumed to be associated with testis development in Chlamys farreri.Therefore,a detailed investigation of this gene was conducted to provide support for a better understanding of testis development in C.farreri.Bioinformatic approaches were used to analyze the sequence characteristics of LAMP1 in C.farreri.Quantitative reverse transcription PCR(qRT-PCR)was employed to examine the expression patterns of LAMP1 in various tissues,including the kidney,hepatopancreas,adductor muscle,gill,mantle,and gonad,as well as across different stages of the gonadal cycle.Additionally,RNA interference(RNAi)was used to investigate the functional role of LAMP1.Bioinformatic analysis revealed that LAMP1 had a coding sequence of 1926 bp,encoding 641 amino acids.The protein contained a highly glycosylated luminal domain,a unique hinge region,a conserved C-terminal transmembrane region,and a short cytoplasmic tail.qRT-PCR revealed differences in LAMP1 expression between sexes and among tissues.In most tissues,male scallops had higher LAMP1 expression than female ones,except in the mantle.Testis expression was 3.27 times greater than ovary expression.In males,the testis exhibited the highest expression,followed by the kidney.In females,the mantle and ovary exhibited relatively high expression levels.RNAi successfully reduced LAMP1 mRNA expression in the experimental group by 65%in ovaries and 62%in testes compared with the blank control and by 62%in ovaries and 60%in testes compared with the negative control.Morphological analysis after RNAi treatment revealed no significant changes in the ovary;however,the testis displayed abnormal sperm morphology.The sperm were short and spherical,unlike the typical elongated shape seen in the controls.LAMP1 is predominantly expressed in the testes of comb-toothed scallops.It is mainly involved in the differentiation of spermatogonia into spermatozoa.This gene is hypothesized to regulate normal sperm morphology.It may facilitate the autophagic clearance of structurally abnormal spermatozoa.