Spermiation is the process by which mature spermatids are released from the supporting somatic Sertoli cells into the lumen of the seminiferous tubule. Aquaporins, the channel proteins which facilitate water and ion exchange, are suggested for possible involvement in the hydration of the seminal fluid as well as in sperm motility. The characterisation of Aquaporin 7 and its expression analysis was carried out in walking catfish (Clarias batrachus). By using primers based on conserved regions of other related species, AQP 7 gene was amplified. With the help of RACE PCR, 5' and 3' end were amplified and PCR product were cloned and sequenced. The sequence results ORF of 972 nucleotide that encodes for a protein of 323 amino acid with no signal peptide. Six transmembrane helices were predicted using Phyre 2 software. In differential tissue expression study in case of female AQP 7 transcripts were found significantly in high quantity in ovary followed by intestine, kidney and gill in quantitative PCR and in case of male higher expression was observed in intestine followed by testes, kidney and gill tissues. Tissue expression study was studied in gonad after post injection of ovatide at intervals of 0 hour, 8 hour, 16 hour and 32 hour. In case of ovary higher expression of AQP 7 was observed at 16 hour while in case of testes 32 hour shows higher expression. In AQP 7 expression shows higher in mature followed by premature and resting breeding season. The findings support the hypothesis of hydration of sperm is aquaporin 7 mediated.