PubMed Health⌕ Search

PubMed · 7309792

Polymorphonuclear leukocyte migration through human amnion membrane.

Abstract

A new in vitro model has been developed for studying migration of human polymorphonuclear leukocytes (PMN) through living native cellular and matrix barriers. Human amnion membrane consists of a single layer of epithelium bound to a continuous basement membrane interfacing an avascular collagenous stroma. Living amnion was placed in plastic chambers with separate compartments on each side of the membrane. PMN were introduced on the epithelial side of the amnion, and a Millipore filter (Millipore Corp., Bedford, Mass.) was placed against the stromal side. In response to N-formylmethionyl-leucyl- phenylanlanine (FMLP) chemoattractant, PMN penetrated the full thickness of the amnion and were collected and counted on the filter. The rate of PMN traversal of the amnion was dependent on the concentration of FMLP (optimal at 10(-8)M) as well as the slope of the FMLP gradient across the amnion. The route of PMN migration was studied by transmission electron microscopy. PMN first attached to the epithelial surface, then infiltrated between intercellular junctions. PMN migrated around or through tight junction and hemidesmosome attachments. The PMN then penetrated the basement membrane and migrated through the dense collagenous stroma. The present amnion migration system has characteristics of the in vivo inflammatory state not described in any previous method for monitoring PMN migration in vitro. Prior methods have not used native epithelium, whole basement membrane, or collagenous stroma. PMN penetration of these barriers occurs in the normal inflammatory response and probably involves biochemical mechanisms not required for simple migration through the pores of an artificial filter. The amnion system can be useful for future biochemical and morphological studies of PMN penetration of these barriers and possible repair processes that may follow.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

R G Russo, L A Liotta, U Thorgeirsson, R Brundage, E Schiffmann. 1981. Polymorphonuclear leukocyte migration through human amnion membrane.. https://doi.org/10.1083/jcb.91.2.459

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

Implantation of human amniotic epithelial cells prevents the degeneration of nigral dopamine neurons in rats with 6-hydroxydopamine lesions.

We recently found that human amniotic epithelial (HAE) cells secrete biologically active neurotrophins such as brain-derived neurotrophic factor and neurotrophin-3, both of which exhibit trophic activities on dopamine (DA) neurons. The present study explored whether implantation of HAE cells can be a possible means to deliver trophic factors into the brain to prevent the death of DA neurons in a rat model of Parkinson's disease. We first investigated the ability of HAE cells to produce factors capable of promoting DA cell survival in vitro, and then tested whether HAE cell grafts survive and prevent the death of nigral DA neurons in rats with 6-hydroxydopamine lesions. A treatment with conditioned medium derived from HAE cell cultures enhanced the survival of tyrosine hydroxylase (TH)-immunopositive DA cells in serum-free cultures. The conditioned medium also protected the morphological integrity of TH-positive neurons against toxic insult with 6-hydroxydopamine. HAE cells were grafted into the midbrain of immunosuppressed rats. The rats were then subjected to a unilateral nigrostriatal lesion induced by intrastriatal infusions of 6-hydroxydopamine. HAE cell transplants were found to survive without evidence for overgrowth 2 weeks postgrafting. The number of nigral DA cells, detected with either TH-immunohistochemistry or retrograde labelling with fluorogold, was significantly increased in rats given the grafts as compared to that in control animals without the grafts. The results indicate that HAE cells produce diffusible molecules that can enhance the survival of DA neurons. Although the factors that contribute to the currently observed effects remain to be fully determined, implantation of HAE cells could be a viable strategy to counteract the loss of DA neurons in Parkinson's disease.

Amnion↗

Evidence of dopamine D1 receptor mRNA and binding sites in cultured human amniotic epithelial cells.

In this study, we employed RT-PCR and radioligand binding studies to evaluate the gene expression and binding characteristics, respectively, of dopamine D(1) receptors in human amniotic epithelial cells (HAEC). The results showed that HAEC natively expressed D(1) receptor mRNA, as measured by RT-PCR, which was identical to that of human brain. Saturation binding studies using [(3)H]SCH 23390 demonstrated the presence of a high affinity D(1) site in HAEC with K(D) and B(max) values of 2.01+/-0.25 nM and 32.5+/-3.7 fmol/mg protein, respectively. Competition studies showed that selective D(1) antagonists were potent displacers of [(3)H]SCH 23390 binding with a potency order consistent with D(1) receptor characteristics. The current results present compelling evidence that HAEC natively express D(1) receptor mRNA and binding sites. The results also establish a primate cell model that can possibly be used for studying D(1) receptor signal transduction and molecular mechanisms and exploring newly developed drugs acting at these receptors.

Amnion↗