Renal Na+, K+-ATPase in SHR: studies of activity and gene expression.
The mechanism by which increased dietary intake of calcium reduces blood pressure in the spontaneously hypertensive rat is unknown. The present studies were designed to determine if there were alterations in the activity of the major membrane ion translocating pump, sodium, potassium-ATPase (NKA), in the kidneys of hypertensive rats and whether increased dietary calcium intake affected the activity of this enzyme. Fifteen-week old SHR's were found to have lower total ATPase activity in microsomal preparations from the kidney than age matched Wistar-Kyoto animals. Both the ouabain-sensitive component (NKA) and the ouabain-insensitive component were lower in SHR. Increasing dietary calcium intake from 1% to 3% elevated both components of the ATPase activity in SHR, but was without effect in WKY. Measurement of membrane phospholipid composition suggested that altered phospholipid composition did not account for the reduced ATPase activity observed, but indicated a reduced density of ATPase in SHR. A technique has been devised for qualitative and quantitative analysis of Na, K-ATPase alpha isoforms using RT-PCR. This technique reveals that the alpha 1 isoform is the sole catalytic isoform present in the nephron. Accurate and precise quantification of the amount of gene expression in individual nephron segments is reported and will be applied to determine whether dietary calcium influences blood pressure by a mechanism which alters nephron NKA gene expression.
Institute of Molecular Medicine, University of Texas Health Science Center, Houston 77030, USA.
A T
AT
Nguyen
A
A
Hayward-Lester
S
S
Sabatini
P A
PA
Doris
D000818:Animals; D001794:Blood Pressure; D002136:Calcium, Dietary / Q000494:pharmacology; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D006973:Hypertension / Q000201:enzymology*; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D010743:Phospholipids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*
D002136:Calcium, Dietary; D017931:DNA Primers; D010743:Phospholipids; D012313:RNA; D000254:Sodium-Potassium-Exchanging ATPase