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C Van Ginneken

Publications and source records attributed to C Van Ginneken.

6 recordsLinked to original sources

A stereological evaluation of secretin and gastric inhibitory peptide-containing mucosal cells of the perinatal small intestine of the pig.

Stereological methods were used to quantify secretin and gastric inhibitory peptide (GIP)-immunoreactivity (GIP-IR) in paraffin sections of the duodenum, jejunum and ileum of fetal and neonatal piglets. In addition, sections were processed for GLP-1-immunohistochemistry. The volume density of the tunica mucosa increased after birth, giving rise to a decreased volume density of the tela submucosa and tunica muscularis. Generally known region-specific morphological distinctions were reflected in differing volume densities of the various layers. The highest volume density of GIP-IR epithelial cells was observed in the jejunum of the neonate. In contrast, the volume density of secretin-IR epithelial cells was highest in the duodenum of both fetal and neonatal piglets. The volume occupied by GIP-IR and secretin-IR epithelial cells increased in the jejunum after birth. Additionally, ileal secretin-IR epithelial cells were more numerous in the neonatal piglet. In conclusion, the quantitative and qualitative presence of GIP-IR and secretin-IR epithelial cells agree with earlier reports of their presence and co-localization between GIP-IR and GLP-1-IR, in the pig small intestine. Furthermore, the differences suggest that age- and region-related functional demands are temporally and probably causally related with the morphological diversification of the intestine and its endocrine cells.

Animals↗

A stereologic evaluation of glucagon-like peptide-1 (GLP-1) mucosal cells in the small intestine of the developing pig.

Studies that investigate possible developmental changes in gastrointestinal hormones in the pig are sparse and contradictory. Therefore, a quantitative morphological study using stereologic methods on paraffin sections of the different small intestinal regions (cranial and caudal duodenum, jejunum and ileum) of the developing pig (second half of the gestation, neonatal and weaning period) has been conducted. The sections were processed for GLP-1-immunohistochemistry. During the investigated time span, the volume of GLP-1-IR cells increased approximately 40-fold in both the jejunum and ileum, notwithstanding a decrease of their volume density. The ileal small intestinal segment contained the highest volume density of GLP-1-IR cells. In contrast, GLP-1-IR cells were only occasionally encountered in the duodenum. The development-related changes of the investigated parameters coincide with dietary changes and the regional differences are in accordance with functional reports that point to GLP-1 as a gastrointestinal hormone that ediates the 'ileal brake' and stimulates glucose-dependent pancreatic insulin secretion.

Animals↗

Stereologic characteristics of pig small intestine during normal development.

Stereologic methods were used to study the behavior of the pig's intestinal wall during periods that are characterized by a high incidence of gastrointestinal disorders. For this purpose conventionally stained transverse and vertical paraffin sections were made of the small intestine (duodenum, jejunum, and ileum) of fetal, neonatal, and weaned pigs. The volumes of the intestinal walls were estimated using Cavalieri's method. Subsequently, the surface density (Sv) of the tunica mucosa and the volume densities (Vv) of the different small intestinal elements were estimated. Finally, the surface and volumes per serosal surface area (Ss and Vs) were calculated. The decrease of Sv can be attributed to the finding that the mucosal surface increases to a lesser extent compared with the volume of the intestinal wall. The Vs of the various layers increased postnatally, illustrating that the intestinal wall thickens. Despite an increasing total mucosal surface, this postnatal thickening causes Ss to decline. Each of these changes is temporally related to dietary changes, an increased antigen load, and an increased need for protection. Additionally, the regional differences of the various parameters match the qualitative descriptions of the small intestine of the pig and relate to region-specific functions.

Animals↗

Developmental changes in heme-oxygenase-2 and bNOS expression in enteric neurons in the pig duodenum.

There exists much parallelism between carbon monoxide- and nitric oxide-generating systems. Therefore, we wondered whether developmental and functional differences along the duodenum similarly affect, part of them, namely, heme oxygenase-2-(HO-2) and neural isoform of nitric oxide synthase- (nNOS) expressing neurons. By applying NADPH diaphorase histochemistry and HO-2 immunohistochemistry on whole-mount preparations and by using stereologic methods, a qualitative and quantitative description of HO-2 and nNOS expression was obtained. Examinations were carried out on the duodenum of fetal, neonatal and weaned pigs. At all ages, three enteric plexuses were readily distinguished. The presence of both enzymes fits in with other morphological and physiological reports. However, the expression of both enzymes significantly changed during development. The number of HO-2-IR neurons increased approximately 20-fold in the inner submucous and almost doubled in the myenteric plexus. In addition, the number of nNOS-expressing neurons displayed a significant decrease in the outer submucous plexus after weaning. High levels of glucocorticoids may cause the perinatally increased HO-2 expression, whereas an influence on nNOS expression is doubtful. Therefore, it seems that notwithstanding the high similarity between both systems, their expression is regulated differently in the pig duodenum.

Animals↗

Nitric oxide synthase expression in enteric neurons during development in the pig duodenum.

The expression of the constitutive neural isoform of nitric oxide synthase (bNOS) is dynamic and thus forms an ideal parameter to evaluate whether development and region affect the enteric nervous system. By applying NADPH-diaphorase histochemistry on whole-mount preparations of the myenteric and submucosal plexuses and by using the 'unbiased counting frame', a qualitative and quantitative description of bNOS-expression in enteric neurons in the pig duodenum in various developmental stage and region was obtained. Examinations were carried out on the oral and aboral duodenum of fetal pigs from the second half of gestation, of 1-2-day-old pigs and of 6-8-week-old pigs. In the pig duodenum, three enteric plexuses were readily distinguished: the inner submucous, the outer submucous and the myenteric plexuses. All three plexuses already harboured, to different degrees, bNOS-expressing neurons at midgestation. Although the enteric nervous system was present at midgestation, the enteric neurons had not yet reached their adult phenotype and morphology. During gestation, the number of inner submucous bNOS-expressing neurons increased approximately 50-fold, whereas after birth that number fell to about 10% of the prenatal value. During further postnatal development it returned to prenatal values. In addition, the number of bNOS-expressing myenteric neurons doubled postnatally. These changes favour a role for NO in mediating the development of enteric neurons and point to a greater necessity for inhibitory innervation in the adult pig as compared with the fetal pig. Furthermore, the number of bNOS-expressing outer submucosal and myenteric neurons was significantly higher in the oral duodenal segment compared with the aboral duodenal segment. This regional difference suggests that the oral duodenal segment is more prominently involved in the regulation of NO-mediated gastrointestinal processes than the aboral one. The developmentally and regionally dependent bNOS-expression can be explained by shifts and differences in the balanced system of hormones, presynaptic input and target-derived signals that affects neurotransmitter expression.

Age Factors↗

Microwave staining of intestinal whole-mount preparations with Cuprolinic blue.

Encouraged by the knowledge that microwaves have a beneficial effect on immunohistochemical reactions, the present study aimed to find out whether microwaves could improve the Cuprolinic Blue staining of enteric neurons as well as the actual method that has been developed for gastrointestinal whole-mount preparations. In addition to incorporating a microwave application in the method described by Holst and Powley (1995), some other modifications were made: two incubations before incubation in the staining solution and free-floating incubations. In the whole-mount preparations, most, if not all, enteric neurons were stained by Cuprolinic Blue. These neurons appeared as blue-green cells with non-reacting nuclei and neuronal processes. At higher magnification, the cytoplasm was characterized by a fine blue-green granulation, and the nucleolus in the nucleus appeared as a blue iridescent structure. Non-specific staining occurred in fibrocytes and epithelial cells but, because of their location and appearance, they could easily be distinguished from neurons. The modified incubations and the incorporation of a microwave application into the conventional Cuprolinic Blue staining method turn the method into an easy-to-use one that seems to visualize most, if not all, enteric neurons in whole-mount preparations of the pig jejunum.

Animals↗