Sequence analysis of hypothalamic parathyroid hormone messenger ribonucleic acid

Endocrinology
M T NutleyS Harvey

Abstract

PTH-like peptides and messenger RNA (mRNA) have recently been detected in neural tissues, but it is uncertain whether this reflects the transcription of the PTH gene or that of a closely related gene. This possibility has, therefore, been investigated. PTH-like complementary DNA (cDNA) moieties of predicted size were readily generated from reverse transcribed brain (hypothalamic and extrahypothalamic tissue) and pituitary RNA, using polymerase chain reaction (PCR) with three sets of overlapping oligonucleotide primers designed to amplify PTH cDNA fragments of 285, 372, and 459 base pairs (bp). PCR reamplification of the largest hypothalamic moiety with an internal set of primers also generated a cDNA fragment of the predicted size (372 bp). Restriction endonuclease digestion with BstNI cleaved the largest hypothalamic cDNA moieties into smaller fragments of 217 and 242 bp, identical to the cleavage of parathyroidal PTH cDNA. Rapid amplification of cDNA ends of the 3'-flanking cDNA sequences also produced hypothalamic and extra-hypothalamic cDNA moieties identical in size (499 bp) to parathyroidal PTH cDNA. Southern analysis of these PCR and rapid amplification of cDNA end cDNA fragments further indicated homology with PTH cDNA....Continue Reading

Citations

May 9, 2001·The Journal of Clinical Investigation·Y ImanishiA Arnold
Apr 17, 2014·Transgenic Research·Ziqiang YuanMichael A Levine
Oct 6, 2000·Journal of Endocrinological Investigation·L M Calvi, E Schipani
Nov 24, 1999·The American Journal of Physiology·M MannstadtT J Gardella

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