L. Berghman

Characterisation of chicken monocytes, macrophages and interdigitating cells by the monoclonal antibody KUL01.

The distribution, function and ontogeny of the mononuclear phagocyte system of the chicken were characterised using the monoclonal antibody (MAb) KUL01. KUL01 specifically recognises chicken monocytes, macrophages and interdigitating cells, as well as activated microglia cells. Its tissue distribution allowed to discriminate KUL01 from all earlier described MAb, reactive with mononuclear phagocytes. The specificity of KUL01 for mononuclear phagocytes was further confirmed in functional assays: KUL01-positive macrophages in spleen and liver actively took up colloidal carbon, while monocytes and spleen and gut macrophages contained non-specific esterase and acid phosphatase activities characteristic for antigen-processing. Further, it was demonstrated that KUL01-reactive peripheral blood monocytes express MHC-II, but not CD4. In all tissues investigated, the same morphological subtypes of macrophages were detected in chicken at similar localisations as in mammals, indicating a high degree of conservation between the mononuclear phagocyte system of the chicken and of mammals.

Immunocytochemistry and immunoblotting of avian prolactins using polyclonal and monoclonal antibodies toward a synthetic fragment of chicken prolactin

A major obstacle in the production of specific antibodies toward chicken prolactin (PRL) has been overcome by mimicking a putative epitope of the molecule using the synthetic decapeptide Lys-chPRL 59-67. This peptide represents the highest hydrophilicity peak of the amino acid sequence of chPRL that was recently derived from the nucleotide sequence. Polyclonal mouse antisera against the fragment specifically recognized the lactotropes in the cephalic lobe of the chicken pars distalis as illustrated by immunocytochemical double staining experiments. Monoclonal antibody production yielded antibodies that specifically labeled purified turkey PRL upon SDS-PAGE separation and immunoblotting. Turkey and chicken PRL showed a very similar polymorphism with respect to their apparent molecular weights, including the occurrence of a glycosylated variant of chicken PRL. The monoclonal antibodies were finally used to demonstrate the presence of PRL-like immunoreactivity both in the pituitary gland and in the brain of the quail. In the brain, immunoreactive neurons were in the nucleus accumbens and in the lateral parts of the ventro-medial hypothalamus, partly similar to those described in the rat.

Immunoaffinity purification and partial characterization of sea bass (Dicentrarchus labrax) growth hormone

Growth hormone (GH) was isolated from sea bass (Dicentrarchus labrax) pituitary extract by a simple one-step procedure involving immunoaffinity chromatography. A monoclonal antibody raised against chicken GH and found to immunostain very specifically the GH cells in the pituitary of the sea bass was coupled to CNBr-activated Sepharose 4B. Sea bass pituitary extracts were run on the affinity column, and the eluted material was analyzed on reversed-phase HPLC and found to consist of one single peak. The yield of purified hormone was 2.4 mg/g pituitary. Two monomeric forms (MW = 20,000 and 22,000 Da) of sea bass GH were identified by gel electrophoresis. Gel electrofocusing revealed apparent isoelectric points of 6.15, 6.50, and 6.95. Amino acid composition is consistent with other vertebrate GHs. The immunological relatedness was tested by immunoblotting using antisera raised against GH of different species. Polyclonal antisera raised against the isolated hormone exhibited a specific labeling of the GH cells in sea bass pituitary sections as well as of the immunoblotted purified GH.

Disappearance rate of glycosylated and non-glycosylated chicken growth hormone: influence on biological activity

The disappearance rates for glycosylated (GcGH) and non-glycosylated chicken growth hormone (NGcGH) were compared following their intravenous injection into anaesthetized adult laying hens. The metabolic clearance rate of GcGH was about 20% lower (P = 0.13) compared to NGcGH. Similarly, there was a tendency towards a longer half-life for GcGH. No major physiological significance could be attributed to the glycosylation state of the preparations as far as their effects on circulating insulin-like growth factor I and iodohormone levels were concerned.

Pulsatility of plasma growth hormone and hepatic growth hormone receptor characteristics of broiler chickens divergently selected for abdominal fat content.

1. Plasma growth hormone (GH) pulsatility and hepatic GH receptor characteristics were compared in experimental lines of meat-type chickens selected for high (HF) or low (LF) abdominal fat content. 2. Mean GH concentration, baseline and amplitude of pulses were slightly, but not significantly, greater in LF chickens. Length and frequency of pulses were similar. 3. LF chickens exhibited higher plasma triiodothyronine (T3) concentrations. This difference between genotypes disappeared when the diet was supplemented with 1 mg/kg T3. 4. Specific binding of GH to liver membranes was higher for the fat line but was depressed by T3 supplementation to the same level in both lines. No difference was observed between lines for affinity constants. 5. It is concluded that direct selection for leanness has a less pronounced, if any, effect on GH pulsatility as compared with selection for food conversion efficiency; therefore, different physiological mechanisms are triggered to achieve leanness.