Comparative studies on Na-dependent Pi transport in ovine, caprine and porcine renal cortex.
In contrast to monogastric species, renal excretion of inorganic phosphate (Pi) in ruminants is low and this could be attributed to an almost complete tubular Pi reabsorption. However, the functional and regulatory basis for this phenomenon has not yet been clarified. Therefore, it was the aim of the present study to characterize the kinetic parameters of the tubular Pi reabsorption system as affected by P or Ca depletion using preparations of renal cortex brush border membrane vesicles (BBMV) from goats and sheep and to compare the data with respective parameters of porcine preparations. Na-dependent Pi uptake into renal cortex BBMV as a function of Pi concentration showed typical Michaelis-Menten kinetic and respective Scatchard plot analysis of the specific Pi uptake revealed linearity indicating the predominant presence of a single type of Pi transporters in the preparations. Under control conditions Vmax values of Na-dependent Pi uptake into BBMV were highest in goats and sheep and lowest in pigs (1.98, 1.39 and 0.95 nmol x mg(-1) protein x 10s(-1), respectively). Km values were not different between goats and sheep under all feeding conditions and ranged from between 0.34 mmol x l(-1) and 0.55 mmol x l(-1) which was three- to five-times higher than that found in pigs (0.11 mmol x l(-1)). Oligonucleotides derived from rat kidney cortex type IIa Na/Pi cDNA were used for reverse transcriptase-polymerase chain reaction (RT-PCR) in goat, sheep and pig kidney cortex. The products isolated were 768 bp for sheep and pigs and 765 bp for goats, with the respective amino acids sequences, representing a segment of approximately 40% in length of the entire transporter, exhibiting an at least 92% sequence homology between different species. From the results, involvement of type IIa Na/Pi cotransport in tubular Pi reabsorption in small ruminants can be postulated. However, it should not be considered that a potential role of other Pi transport systems be completely be excluded. Interestingly, neither P nor Ca depletion caused significant effects on Na-dependent Pi transport capacities and affinities in goats and sheep. From this, parathyroid-hormone independent regulatory pathways of tubular Pi reabsorption can be assumed.
Schröder
B
B
Department of Physiology, School of Veterinary Medicine, Hannover, Germany. bschroed@physiology.tiho-hannover.de
Walter
C
C
Breves
G
G
Huber
K
K
D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary; D015870:Gene Expression / Q000502:physiology; D006041:Goats; D007672:Kidney Cortex / Q000378:metabolism*; D008297:Male; D008871:Microvilli / Q000378:metabolism; D008969:Molecular Sequence Data; D010710:Phosphates / Q000378:metabolism*; D051381:Rats; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D012964:Sodium / Q000378:metabolism*; D050608:Sodium-Phosphate Cotransporter Proteins; D050613:Sodium-Phosphate Cotransporter Proteins, Type IIa; D013045:Species Specificity; D013552:Swine; D027981:Symporters
D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D010710:Phosphates; C493556:Slc34a1 protein, rat; D050608:Sodium-Phosphate Cotransporter Proteins; D050613:Sodium-Phosphate Cotransporter Proteins, Type IIa; D027981:Symporters; D012964:Sodium; D002118:Calcium