Detection of mRNA encoding guanylate cyclase A/atrial natriuretic peptide receptor in the rat cochlea by competitive polymerase chain reaction and in situ hybridization.
Expression of mRNA encoding guanylate cyclase A (GC-A)/atrial natriuretic peptide (ANP) receptor in the rat cochlea was examined by polymerase chain reaction (PCR) and in situ hybridization (ISH). After reverse-transcription, PCR amplification, subcloning, and sequencing, we found that GC-A mRNA with sequence identical to that previously cloned from the rat brain (Chinkers et al., 1989) was expressed in the rat spiral ligament as well as in the spiral ganglion. However, GC-A mRNA was not detected in the stria vascularis. Competitive PCR using internal standard DNAs indicated that the expression of GC-A in the cochlea occurred at a level approximately 16 times less than that measured in kidney cortex. ISH histochemistry using a 35S-labeled antisense riboprobe showed the highest level of expression of GC-A mRNA to be in oligodendrocytes of the cochlear nerve root. The results suggest that ANP may play a role in the cochlear nerve function.
Furuta
H
H
Department of Surgery/Otolaryngology, UCSD, School of Medicine, La Jolla, 92093-0666, USA.
Mori
N
N
Luo
L
L
Ryan
A F
AF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003051:Cochlea / Q000201:enzymology / Q000378:metabolism*; D003056:Cochlear Nerve / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D006162:Guanylate Cyclase / Q000235:genetics* / Q000378:metabolism; D017403:In Situ Hybridization; D007672:Kidney Cortex / Q000201:enzymology / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013136:Spiral Ganglion / Q000378:metabolism; D013316:Stria Vascularis / Q000378:metabolism
D017931:DNA Primers; D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor