Decreased tyrosine hydroxylase mRNA but not cholecystokinin mRNA in the pars compacta of the substantia nigra and ventral tegmental area of aged rats.
Quantitative in situ hybridization histochemistry was used to determine the age-related changes in tyrosine hydroxylase (TH) mRNA and cholecystokinin (CCK) mRNA in the ventral tegmental area (VTA) and substantia nigra pars compacta (SNc) of the rat. Coronal sections (10 microns) were cut in a cryostat through the VTA and SNc of brains from 3 months and 33 month old Sprague-Dawley rats and immediately adjacent sections hybridized with 35S-labelled 45-mer oligonucleotide probes specific for either the rat TH or CCK genes. The mRNA levels of each gene were estimated by computerised densitometric analysis of the signal on X-ray film autoradiograms and estimation of the number of mRNA expressing cells as well as the density of expression per cell (grain density) was made from high resolution emulsion autoradiograms. Analysis of the TH mRNA on X-ray film autoradiograms indicated that the levels averaged 25% lower in the SNc (P < 0.01) and 18% lower in the VTA (P < 0.05) of the old rats. However, analysis of the emulsion autoradiograms showed that this reduction in TH mRNA in the VTA and SNc in the old rats was not due to a loss of TH mRNA expressing cells but due to a reduction in the hybridization signal per expressing cell.(ABSTRACT TRUNCATED AT 250 WORDS)
Schuligoi
R
R
Merck Sharp & Dohme Research Laboratories, Neuroscience Research Centre, Harlow, Essex, UK.
Fernandez
J
J
Heavens
R P
RP
Sirinathsinghji
D J
DJ
D000375:Aging / Q000502:physiology*; D000818:Animals; D001345:Autoradiography; D002766:Cholecystokinin / Q000096:biosynthesis*; D015870:Gene Expression / Q000502:physiology*; D017403:In Situ Hybridization; D008297:Male; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013378:Substantia Nigra / Q000254:growth & development / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis*; D017557:Ventral Tegmental Area / Q000254:growth & development / Q000378:metabolism*
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D002766:Cholecystokinin; D014446:Tyrosine 3-Monooxygenase