title: Assessment of the labelling index of cohorts of the pancreatic islet cell population in phenobarbitone-treated male rats using a double immunohistochemical technique for 5-bromo-2'-deoxyuridine and pancreatic hormones.
abstract: A previous study demonstrated that administration of phenobarbitone to male AP Wistar rats for up to 7 days caused alterations in labelling indices (LIs) in several different tissues (including a reduction of the endocrine pancreas population LI) as determined by immunohistochemical visualisation of 5-bromo-2'-deoxyuridine (BrdU) incorporation into S-phase nuclei. The primary objective of this study was to determine whether treatment with phenorbarbitone influenced the replicative states of specific cohorts of the islet (of Langerhans) cell population or generated a uniform depression of LI. Quantitation of the LIs of individual islet cell cohorts was achieved by utilisation of a dual immunohistochemical staining method for BrdU and islet hormones (insulin, glucagon and somatostatin) using a sequential peroxidase anti-peroxidase (PAP)/alkaline phosphatase anti-alkaline phosphatase (APAAP) method employing diaminobenzidine and New Fuchsin chromogens, respectively. We observed reductions, increases and no change in LIs of insulin-, glucagon- and somatostatin-positive cells, respectively. We conclude that the decreased LI of the insulin-positive cohort was not countered entirely by the LI increase in the glucagon-positive cohort due to the larger size of the former. Furthermore, the effects of phenobarbitone treatment are not manifested generally in the islet cell population but in the insulin- and glucagon-positive cohorts only. The causation of these effects is unknown but is likely to be due to enhanced carbohydrate and hormone metabolism. We believe that the visualisation and quantitation of replicating cells in specific hormone-positive cohorts of the islet cell population provide opportunities for understanding the influence of xenobiotics and disease processes on pancreatic function.
authors.affiliation: Experimental Morphology, Safety of Medicines Department, Zeneca Pharmaceuticals, Macclesfield, Cheshire, United Kingdom.
authors.forename: H B
authors.initials: HB
authors.lastname: Jones
authors.forename: S J
authors.initials: SJ
authors.lastname: Harbottle
authors.forename: A L
authors.initials: AL
authors.lastname: Bowdler
mesh_terms: D000469:Alkaline Phosphatase / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000378:metabolism*; D004261:DNA Replication / Q000187:drug effects; D005934:Glucagon / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D010634:Phenobarbital / Q000008:administration & dosage / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D013004:Somatostatin / Q000378:metabolism
chemical_list: D000906:Antibodies; D007328:Insulin; D013004:Somatostatin; D005934:Glucagon; D000469:Alkaline Phosphatase; D001973:Bromodeoxyuridine; D010634:Phenobarbital