Expression patterns of voltage-dependent calcium channel alpha 1 subunits (alpha 1A-alpha 1E) mRNA in rat retina.
The transcript levels of the genes encoding for the different alpha1 (alpha1A-alpha1E) subunits of voltage-dependent calcium channels (VDCCs) were studied in the retina of the rat using RT-PCR, Northern blotting, and in situ hybridization. Abundant expression of alpha1A and alpha1B was found with RT-PCR and on Northern blots of total retina RNA, corresponding with high expression levels in all nuclear layers (outer and inner nuclear layers and the ganglion cell layer) of the retina. VDCC alpha1D mRNA was also present in all nuclear layers of the retina but at less abundant levels than alpha1A or alpha1B. Expression level of alpha1C in the retina was low as deduced from a faint Northern blot signal and a moderate yield after PCR amplification. VDCC alpha1E specific amplification of retinal cDNA yielded a longer product (designated alpha1E-L) than obtained from the hippocampus. Nucleotide sequencing of this PCR product revealed a 129 bp insert which is largely homologous (97%) with a previously described insert in the same position in human alpha1E cDNA. In situ hybridization in rat brain showed a differential expression pattern of the long and short variants of alpha1E mRNA. Northern blotting of retinal RNA confirmed the absence of the short variant (alpha1E-S), while alpha1E-L was present at low levels. In situ hybridization detected a significant level of expression of alpha1E-L in the inner nuclear layer. The prevalent expression of alpha1A and alpha1B, and to a lesser extent, of alpha1D, indicates that P/Q-, N-, and L-type calcium currents play a prominent role in the various cell types involved in the retinal signal-transduction pathway. The absence of alpha1C transcript in the retina suggests that the slowly inactivating L-type calcium currents involved in neurotransmitter release from the terminals of photoreceptors and bipolar cells may be encoded by the alpha1D isoform.
Kamphuis
W
W
Graduate School for the Neurosciences Amsterdam, Netherlands Ophthalmic Research Institute, Department of Ophthalmo-Morphology, PO Box 12141, 1100 AC Amsterdam, Netherlands. w.kamphuis@ioi.knaw.nl
Hendriksen
H
H
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000096:biosynthesis* / Q000235:genetics; D020864:Calcium Channels, N-Type; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015220:Calcium Channels; D020864:Calcium Channels, N-Type; D020033:Protein Isoforms; D012333:RNA, Messenger