Friedman P A
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03756.
Am J Physiol. 1988 Jan;254(1 Pt 2):F62-70. doi: 10.1152/ajprenal.1988.254.1.F62.
These studies examined transepithelial routes of basal and hormone-activated calcium absorption in thick ascending limbs of Henle's loop perfused in vitro. Single cortical (cTAL) or medullary (mTAL) thick ascending limbs were dissected from kidneys of normal mice. Tubules were perfused at 37 degrees C at pH 7.4 in bicarbonate-buffered media for cTALs and in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid-buffered solutions for mTALs. Calcium was determined by electrothermal atomic absorption spectrophotometry. External driving forces for calcium were eliminated by perfusing and bathing tubules with symmetric solutions containing identical calcium concentrations and by abolishing the transepithelial voltage (VT), by adding furosemide or bumetanide to the luminal perfusate. Basal calcium absorption averaged 60 X 10(-12) mol.s-1.cm-2 in cTAL. After addition of 1 U/ml parathyroid hormone (PTH)-(1-34), net calcium absorption increased to 162 X 10(-12) mol.s-1.cm-2 (P less than 0.05) with no change in VT. Similarly, addition of 10(-3) M dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP) to external bathing solutions increased net calcium absorption to 201 from 70 X 10(-12) mol.s-1.cm-2 in control conditions (P less than 0.025). These results demonstrate that PTH, acting at least in part through cAMP, activates a cellular pathway for calcium absorption in cTAL. In segments of mTAL, calcium absorption was strictly voltage dependent: when VT was abolished, net calcium absorption fell to values indistinguishable from zero and was not altered in the presence of dibutyryl cAMP.(ABSTRACT TRUNCATED AT 250 WORDS)
这些研究检测了体外灌注的亨氏袢厚升支中基础钙吸收和激素激活的钙吸收的跨上皮途径。从正常小鼠的肾脏中分离出单个皮质(cTAL)或髓质(mTAL)厚升支。对于cTAL,在37℃、pH 7.4的碳酸氢盐缓冲培养基中灌注小管;对于mTAL,则在N-2-羟乙基哌嗪-N'-2-乙磺酸缓冲溶液中灌注。通过电热原子吸收分光光度法测定钙含量。通过用含有相同钙浓度的对称溶液灌注和浸泡小管,并通过向管腔灌注液中添加呋塞米或布美他尼消除跨上皮电压(VT),消除钙的外部驱动力。cTAL中基础钙吸收平均为60×10⁻¹² mol·s⁻¹·cm⁻²。添加1 U/ml甲状旁腺激素(PTH)-(1-34)后,净钙吸收增加到162×10⁻¹² mol·s⁻¹·cm⁻²(P<0.05),VT无变化。同样,在外部浸泡溶液中添加10⁻³ M二丁酰腺苷3',5'-环磷酸(cAMP),可使对照条件下的净钙吸收从70×10⁻¹² mol·s⁻¹·cm⁻²增加到201(P<0.025)。这些结果表明,PTH至少部分通过cAMP起作用,激活了cTAL中钙吸收的细胞途径。在mTAL段,钙吸收严格依赖电压:当VT消除时,净钙吸收降至与零无差异的值,且在二丁酰cAMP存在时无变化。(摘要截断于250字)