Differential distribution of the vasopressin V receptor along the rat nephron during renal ontogeny and maturation.
BACKGROUND
Ontogeny and cellular distribution of vasopressin receptors in the kidney are key factors determining the role of vasopressin in renal physiology. Expression of vasopressin V(2) receptor (V(2)R) mRNA and the immunoreactive protein in rat kidney were investigated.


METHODS
An antiserum directed to epitope TLD25 of the rat V(2)R sequence was characterized by Western blotting. Expression of V(2)R mRNA was assessed by reverse transcription-polymerase chain reaction (RT-PCR), and on protein level by immunohistochemistry.


RESULTS
Specificity of the antiserum was documented by Western blots from cells expressing a fusion protein of V(2)R and GFP. Using lysates of rat kidney and of native cell lines expressing V(2)R but not V(1)R, our antiserum to peptide TLD25 revealed a major band of 55 kD corresponding to the monomeric form of V(2)R, and a band of 110 kD most likely representing the homodimeric form of the receptor. This highly specific antiserum allowed us to localize the V(2)R in thick ascending limbs, distal convoluted and connecting tubules, and in collecting ducts. During ontogeny, immunoreactivity was first observed at the luminal membrane on prenatal day 20, emerging at the basolateral side from postnatal day 5 on. RT-PCR demonstrated V(2)R transcripts from prenatal day 18 to gradually increasing thereafter.


CONCLUSIONS
Expression of V(2)R is first detectable in the late embryonic stage of rat ontogeny starting from day E18 and gradually increasing with kidney maturation. In the adult kidney, V(2)R is differentially distributed in the various nephron segments.
Sarmiento
José M
JM
Institutos de Fisiología and Histología y Patología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, Chile.
Ehrenfeld
Pamela
P
Añazco
Carolina C
CC
Reyes
Carlos E
CE
Troncoso
Silvia
S
Figueroa
Carlos D
CD
Müller-Esterl
Werner
W
González
Carlos B
CB
D000367:Age Factors; D000818:Animals; D000918:Antibody Specificity; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005865:Gestational Age; D007150:Immunohistochemistry; D007685:Kidney Tubules, Collecting / Q000196:embryology / Q000502:physiology; D007686:Kidney Tubules, Distal / Q000196:embryology / Q000502:physiology; D008138:Loop of Henle / Q000196:embryology / Q000502:physiology; D008297:Male; D009399:Nephrons / Q000196:embryology* / Q000502:physiology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017483:Receptors, Vasopressin / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
D012333:RNA, Messenger; D017483:Receptors, Vasopressin