Volume 117, Issue 1, Pages 191-199 (July 1999) Cholinergic system modulates growth, apoptosis, and secretion of cholangiocytes from bile duct–ligated rats  Gene LeSage*, Domenico Alvaro‡, Antonio Benedetti§, Shannon Glaser∥, Luca Marucci§, Leonardo Baiocchi¶, Wendy Eisel#, Alessandra Caligiuri*, Jo Lynne Phinizy∥, Rebecca Rodgers¶, Heather Francis∥, Gianfranco Alpini*,¶,#  Gastroenterology  Volume 117, Issue 1, Pages 191-199 (July 1999) DOI: 10.1016/S0016-5085(99)70567-6 Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 1 Western blot analysis (representative blot of 2 experiments) of muscarinic receptors in cholangiocytes from BDL or BDL+vagotomy rats. The intensity of the muscarinic receptor band was 10-fold higher in cholangiocytes from BDL rats than in cholangiocytes from BDL+vagotomy rats. Gastroenterology 1999 117, 191-199DOI: (10.1016/S0016-5085(99)70567-6) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 2 Immunohistochemistry for CK-19 in frozen liver sections (n = 6) from (A) BDL and (B) BDL+vagotomy rats. Note that vagotomy induced a marked decrease in the number of ducts compared with BDL control rats (original magnification 125×). (C) Liver section from a BDL+vagotomy rat stained with H&E. Two apoptotic bodies (arrows) appear in the epithelium of a bile duct (arrowhead) that is surrounded by hepatocytes and located in proximity of an enlarged portal space (representative section of 6 examined). (D) Rat liver section 7 days after BDL, vagotomy, and forskolin treatment showing proliferating bile ducts without any evidence of apoptosis. This was seen in the portal spaces examined (representative section of 6 examined; original magnification 100×). Gastroenterology 1999 117, 191-199DOI: (10.1016/S0016-5085(99)70567-6) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 3 (A) Gene expression for PCNA (■) and H3-histone (2) was determined by direct RNase protection assay. Autoradiograms (n = 4) were quantified by densitometry. *P < 0.05 vs. cholangiocytes from BDL rats. (B) Effect of long-term administration of forskolin or DMSO on the expression of PCNA (■) and H3-histone (2) mRNAs in cholangiocytes from BDL+vagotomy rats. Forskolin increases the expression of both PCNA and H3-histone mRNAs compared with cholangiocytes from DMSO-treated rats. Autoradiograms (n = 6) were quantified by densitometry. *P < 0.05 vs. cholangiocytes from DMSO-treated BDL+vagotomy rats. Gastroenterology 1999 117, 191-199DOI: (10.1016/S0016-5085(99)70567-6) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 4 (A) SR gene expression in cholangiocytes from BDL and BDL+vagotomy rats was measured by direct RNase protection assay. Autoradiograms (n = 4) were quantified by densitometry. *P < 0.05 vs. basal values. (B) Effect of long-term administration of forskolin or DMSO (control) on SR gene expression in cholangiocytes from BDL+vagotomy rats. Forskolin administration increases the expression of SR mRNA in purified cholangiocytes compared with cholangiocytes from DMSO-treated controls. Autoradiograms (n = 6) were quantified by densitometry. *P < 0.05 vs. corresponding basal values. Gastroenterology 1999 117, 191-199DOI: (10.1016/S0016-5085(99)70567-6) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 5 (A) Measurement of basal (■) and secretin (10−7 mol/L; 2)-induced cAMP levels in cholangiocytes from BDL or BDL+vagotomy rats. *P < 0.05 vs. basal cAMP levels of cholangiocytes from BDL rats. **P < 0.05 vs. secretin-induced cAMP levels of cholangiocytes from BDL rats. Data are means ± SE of at least 6 values. (B) Effect of long-term administration of forskolin or DMSO on basal (■) and secretin (10−7 mol/L; 2)-induced cAMP synthesis in cholangiocytes from BDL+vagotomy rats. Forskolin administration increases cAMP levels in cholangiocytes from BDL+vagotomy rats compared with cholangiocytes from DMSO-treated controls. *P < 0.05 vs. basal cAMP levels. **P < 0.05 vs. secretin induced cAMP levels of cholangiocytes from BDL+vagotomy rats treated with DMSO. Data are means ± SE of at least 10 values. Gastroenterology 1999 117, 191-199DOI: (10.1016/S0016-5085(99)70567-6) Copyright © 1999 American Gastroenterological Association Terms and Conditions