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Kenji Machida

Publications and source records attributed to Kenji Machida.

5 recordsLinked to original sources
PubMed2006

Downregulation of the V2 vasopressin receptor in dehydration: mechanisms and role of renal prostaglandin synthesis.

The vasopressin-aquaporin 2 system plays a key role in urine concentration in dehydration. In contrast to the upregulation of aquaporin 2, the downregulation of the vasopressin V2 receptor in dehydration is known. We investigated the mechanisms of this downregulation in dehydration using reverse transcription-competitive polymerase chain reaction (RT-competitive PCR) and Western blot analysis. The incubation of microdissected inner medullary collecting ducts (IMCDs) in a hypertonic medium or with vasopressin stimulated V2 receptor mRNA and protein expression, showing that dehydration-induced hyperosmolality in renal medulla and increased plasma arginine vasopressin (AVP) concentration should upregulate V2 receptor. The presence of inhibitory factors on the V2 receptor in dehydration was suggested. Prostaglandin E(2) (PGE(2)) is known to inhibit AVP-induced cAMP production and to increase production in dehydration. PGE(2) slightly stimulated V2 receptor mRNA expression in IMCD in vitro. However, PGE(2) inhibited V2 receptor mRNA expression in IMCD in the presence of 10(-9) M vasopressin. The blockade of PGE(2) synthesis by indomethacin in dehydrated rats increased V2 receptor protein expression after 24-48 h with an early increase in V2 receptor mRNA expression. In summary, these data suggest that increased production of PGE(2) in renal medulla plays a key role in the downregulation of V2 receptor in dehydration.

Animals↗
PubMed2003

Mechanisms of down-regulation of the renal parathyroid hormone receptor in rats with chronic renal failure.

Hypocalcemia, hyperphosphatemia and resistance to the action of parathyroid hormone (PTH) are well-characterized features in advanced chronic renal failure (CRF). Their pathogenesis has been attributed to both PTH receptor (PTH-R) down-regulation and postreceptor abnormalities. In this study, we examined the renal expression of the PTH-R mRNA in CRF (5/6 nephrectomy) rats. Experiments were also performed to determine whether an acidic condition and PTH itself influence PTH-R mRNA expression. RT-competitive PCR was used to examine mRNA expression, and polyclonal antibody against PTH-R was used for Western blot. PTH-R mRNA expression was abundant in glomeruli, proximal convoluted and straight tubules (PCT, PST), small in medullary and cortical thick ascending limbs, and cortical collecting ducts and not detectable in outer and inner medullary collecting ducts. The expression was significantly decreased in PCT and PST in CRF rats. Decrease in PTH-R mRNA expression was observed 1 week after the induction of CRF. PTH-R protein was decreased at 2 (-23%) and 4 (-45%) weeks in renal cortex, but not in medulla in CRF rats. PTH-R mRNA expression in PST was decreased by low pH (7.1 or 6.7) incubation compared with that at pH 7.4. PTH(1-34) (10(-9) M) increased PTH-R mRNA expression in PST from control rats by 250%. The stimulatory effect of PTH on PTH-R mRNA expression was decreased by the incubation at low pH medium. In summary, renal PTH-R is down-regulated in CRF rats. The decrease in mRNA expression in PCT and PST causes the decrease in PTH-R protein. Metabolic acidosis may participate in the down-regulation of PTH-R in early stage of CRF. This abnormality could be important in the pathogenesis of secondary hyperparathyroidism of CRF.

Acidosis↗
PubMed2003

Type 1A angiotensin II receptor is regulated differently in proximal and distal nephron segments.

Angiotensin II plays important roles in renal vasoconstriction, sodium reabsorption in proximal tubules, and cell proliferation. Angiotensin II receptors are present not only in proximal but also in distal tubules. We investigated the effects of dehydration on the mRNA expression of type 1A angiotensin II receptor (AT1A) in proximal and distal nephron segments and on the expression of type 1 angiotensin II receptor (AT1) protein. Competitive polymerase chain reaction was employed to quantitatively examine mRNA expression, and AT1-specific polyclonal antibody was used for Western blot analysis. AT1A mRNA expression was most abundant in glomeruli. Collecting ducts showed higher expressions than did proximal tubules or thick ascending limbs. Dehydration caused an increase of AT1A mRNA expression in glomeruli, proximal straight tubules (PST), and medullary and cortical thick ascending limbs (MAL and CAL, respectively). In contrast, dehydration decreased AT1A mRNA expression in cortical, outer medullary, and inner medullary collecting ducts (CCD, OMCD, and IMCD, respectively). Incubation of isolated glomeruli, PST, and IMCD in hypertonic solution made by NaCl and mannitol in vitro increased AT1A mRNA expression. Incubation of IMCD with AVP (10(-7) mol/l) also increased AT1A mRNA expression. AT1 was detected at 45 kDa by Western blotting. Dehydration caused a decrease and increase of AT1 expression in the cortex and the medulla, respectively. In summary, these data showed that the mechanisms of the regulation of AT1A differ between proximal and distal tubules. The finding that AT1 was up-regulated in the medulla during dehydration may suggest that this receptor plays an important role in dehydration in the distal tubules.

Animals↗
PubMed2003

Acute regulation of the epithelial sodium channel gene by vasopressin and hyperosmolality.

The amiloride-sensitive epithelial sodium channel (ENaC) plays a key role in sodium reabsorption in the collecting ducts. We examined ENaC mRNA distribution along the nephron and acute effects of vasopressin and hyperosmolality on ENaC mRNA expression. ENaCalpha, beta, and gamma mRNA expressions were observed in cortical, outer medullary and initial inner medullary collecting ducts (CCD, OMCD and ilMCD, respectively). ENaCalpha mRNA expression was also observed in medullary and cortical thick ascending limbs (MAL and CAL, respectively), while ENaCbeta and gamma mRNA expressions were not observed. Furthermore, ENaCalpha mRNA expression in MAL but not in collecting ducts was stimulated by acute exposure to arginine vasopressin (AVP), 8-(4-chlorophenylthio) (CPT)-cAMP and hyperosmolality. However, the physiological significance of these effects is not known, since ENaC protein is reported to be absent in MAL. These data suggest that ENaCalpha mRNA expression in MAL but not in collecting ducts is acutely regulated by AVP and hyperosmolality. The absence of stimulation of ENaCalpha mRNA expression in collecting ducts suggests the physiological significance of ENaCbeta and gamma mRNA for acute regulation by vasopressin. Determining the physiological significance of the acute effect of vasopressin in MAL will require further investigations.

Animals↗