Chymotrypsin gene expression in rat peripheral organs

Abstract

Prior studies have revealed the presence of chymotrypsinlike protease in peripheral organs, although no definitive evidence for the synthesis of this enzyme in tissue other than the pancreas is available. In an attempt to detect chymotrypsinogen mRNA in peripheral organs, a fragment of the pancreatic chymotrypsin mRNA from rat was amplified using PCR. The sequence was identified as a portion of the rat chymotrypsin B gene overlapping exon 5 through exon 7. It was subcloned into the pGEM-4Z vector and used as a template for the vitro transcription of an antisense riboprobe. Using ribonuclease protection and Northern blot analyses, chymotrypsin mRNA was detected in the rat pancreas, stomach, duodenum, ovary, and spleen. Monoclonal and polyclonal antisera against chymotrypsin detected chymotrypsinlike immunoreactivity in rat and human pancreas, rat stomach, duodenum and jejunum. Electrophoresis and immunoblotting revealed chymotrypsin-chymotrypsinogen bands (25-29 kDa) in the stomach and duodenum. Synthesis of a potent protease such as chymotrypsin in tissue other than pancreas is significant, suggesting a potential physiological and/or pathological role in these tissues.

Impact of developmental lead exposure on splenic factors

“…In other studies, serum levels of pancreatic amylase have been observed to increase due to various forms of stress (Kubisch and Logsdon, 2007; Cosen-Binker et al, 2004; Takano et al, 1992). Up-regulation of chymotrypsin, elastase, carboxypeptidases and trypsin 4 by Pb could be indications that Pb is causing inflammation or injury to the spleen, since proteases have been found to be released from lysosomes during inflammatory reactions (Vischer and Bertrand, 1976; Wang et al, 1998). In addition to activation of the complement system, these proteases can function in the activation and proliferation of lymphocytes, as was demonstrated by Vischer and Bertrand (1976) and Waldo et al (2003).…”

Transcriptional Profiling of Laser Capture Microdissected Rat Arterial Elements: Fenoldopam-induced Vascular Toxicity as a Model System

“…These data support that gross contamination of mesenteric tissue with pancreatic RNA was highly unlikely. In addition, some enzymes primarily considered to be pancreatic-specific have been identified in EC or other tissues and include pancreatic-type carboxyl ester lipase (Li and Hui, 1998), chymotrypsin (Wang et al, 1998), trypsinogen 1 (Koshikawa et al, 1994), chymotrypsin (Wang et al, 1998), and pancreatic carboxyl ester lipase (Shamir et al, 1996). In addition, Koshikawa et al (1997) have reported expression of trypsino-gen-2, a proteinase involved in blood coagulation, angiogenesis, and control of blood pressure, in human vascular EC around gastric tumors and in patients with disseminated intravascular coagulation.…”