Analysis of the cellular expression pattern of beta-CGRP in alpha-CGRP-deficient mice

In this study we compared the alpha-calcitonin gene-related peptide (alphaCGRP) and betaCGRP expression patterns in wild-type and knockout mice by using quantitative reverse transcriptase polymerase chain reaction and immunohistochemistry. In dorsal root ganglia and spinal cord of wild-type animals, alphaCGRP mRNA was about two times more abundant than betaCGRP mRNA. The betaCGRP mRNA was the only isoform expressed in the intestine. In alphaCGRP knockout mice, we found no change in betaCGRP mRNA levels in dorsal root ganglia and spinal cord compared with wild-type controls, but a twofold decrease in the intestine. CGRP immunoreactivity (IR) was detected in many small and some large neurons in the dorsal root ganglia, was found in sensory fibers and motor neurons in the spinal cord, and labeled neuromuscular junctions in wild-type mice. In the dorsal root ganglia of alphaCGRP knockout mice, punctate betaCGRP-IR again was predominantly found in small neurons. In the spinal cord, betaCGRP-IR fibers were localized to the outermost layer of the dorsal horn. IR was found in the cell bodies of motor neurons, but it was undetectable in neuromuscular junctions. In the intestine, CGRP-IR was localized to neurons of the myenteric plexus and to fibers in the mucosal folds, with similar staining intensity in both wild-type and knockout mice. Finally, CGRP-IR was undetectable in preganglionic fibers and postganglionic sympathetic neurons in mice from both genotypes. Our results indicate that alphaCGRP and betaCGRP are variably coexpressed in different functional aspects of the mouse nervous system. This pattern suggests distinct roles for betaCGRP in pain, neuromuscular, and gastrointestinal systems.