CRISP (Cysteine Rich Secretory Proteins) as novel regulators of epididymal epithelium differentiation

CUASNICÚ PS; BATTISTONE MA; CARVAJAL G; LUSTIG L; BRUKMAN NG; BRETON S; WEIGEL MUÑOZ M

Buenos Aires

Congreso; XIX Reunión Anual de la Sociedad Argentina de Biología (SAB). Reunión Conjunta de Sociedades de Biociencias; 2017

Epididymal CRISP1 andCRISP4 associate with sperm during maturation and participate in differentstages of fertilization. Whereas single knockout (KO) males for these moleculesexhibit in vitro sperm fertilizing defects and normal fertility, theabsence of both proteins (double KO, DKO) affects both in vitrofertilization and male fertility, and triggers epididymal inflammation, suggestingthe existence of compensatory mechanisms between these proteins at both the reproductive and immunological level.Whereas the role of these CRISP in fertilization wasthoroughlystudied, their involvement in epididymal physiology andimmunology was novel and unexpected. Based on this, we investigated themechanisms underlining the DKO inflammatory phenotype.Histological studies ofDKO epididymidesshowed the presence of desquamation of the epithelium,cytoplasmic vacuolation and abnormal presence of immune cells in theinterstitium and lumen, in one third of examined males.In addition, intraluminalpH was higher in the affected epididymides and sperm viability decreasedbetween caput and cauda. RT-qPCR for different immunomodulator moleculesrevealed normal levels of Ido but anupregulation of Il-6, Il-10 and a downregulation of Tgf-β only in inflamedepididymides.However, DKO epididymides without signs of inflammation were indistinguishablefrom wildtypes (WT) for these parameters. Immunofluorescence experiments usingspecific markers for different epididymal epithelial cells revealed an immaturephenotype,similarto a pre-pubertal animal, in both inflamed and normal epididymides, suggesting defectspriorto the immunological response and the inflammatory process that may also affectsperm maturation.In this sense, a high percentage of sperm with cytoplasmicdroplet and deficiencies in different motility parameters were found in spermfrom non-inflamed DKO epididymides. Altogether,this work revealed the importance of CRISP inepididymal physiology to preserveits integrity and functionality