Effects of dietary phosphorus and myo-inositol supplementation on NaPi-IIb and TRPV6 protein expression in duodenal apical membranes of laying hens from two strains

Phosphorus (P) and calcium (Ca) absorption in the intestine is mediated by apical brush border membrane (BBM) transporters, including the sodium-dependent phosphate (P_i) transporter NaPi-IIb and the Ca²⁺-selective channel TRPV6. Both are highly expressed in the duodenum and exhibit dietary adaptability; yet little is known about how this adaptability varies with strain and age in laying hens. The present study examined the effects of dietary mineral P renunciation and myo-inositol (MI) supplementation on NaPi-IIb and TRPV6 protein expression in the duodenal BBM of Lohmann Brown-Classic (LB) and Lohmann LSL-Classic (LSL) hens. Two independent feeding trials were conducted: hens received diets either with or without mineral P supplementation (wk 15 - 19 and 20 - 24), or with graded MI levels (0 - 3 g/kg; wk 26 - 30). At the end of each period, hens were euthanized and protein expression of NaPi-IIb and TRPV6 in duodenal BBM was studied by western blotting. Statistical correlation with additional traits of mineral metabolism was analyzed. An immunoreactive NaPi-IIb band was detected at ∼ 45 kDa; therefore, all results reported here refer to this NaPi-IIb fragment. Mineral P renunciation did not affect NaPi-IIb fragment or TRPV6 expression in either hen strain. In LSL hens NaPi-IIb fragment expression increased from wk 19 to wk 24, whereas in LB hens it remained unchanged. NaPi-IIb fragment expression was positively associated with duodenal phosphatase activity and plasma estradiol. TRPV6 expression tended to reduce in LB hens from wk 19 to wk 24, but remained stable in LSL hens. TRPV6 expression was positively associated with duodeno-jejunal P content. MI supplementation upregulated NaPi-IIb fragment expression in LB, but downregulated it in LSL hens with high dietary MI levels, without affecting TRPV6. These findings demonstrate strain-dependent regulatory patterns of duodenal expression of NaPi-IIb fragment and TRPV6 in response to physiological stage and MI supply, indicating that mineral feeding strategies may benefit from genotype-specific consideration, whereas the mechanisms underlying MI-related effects require further clarification.